Most recent papers:

• The role of microRNAs involved in PI3‐kinase signaling pathway in colorectal cancer.
  Saeed Noorolyai, Ahad Mokhtarzadeh, Elham Baghbani, Milad Asadi, Amir Baghbanzadeh Kojabad, Mahsa Maleki Mogaddam, Behzad Baradaran.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- microRNAs have a very crucial role in tumorgenesis and prevention of cancer, which play a significant role in influencing various factors through different signaling pathways. microRNAs can act as tumor suppressors and oncomirs in this cancer through phosphoinositide 3‐kinase/AKT signaling pathway. Abstract In recent decades, cancer has been one of the most important concerns of the human community, which affects human life from many different ways, such as breast, lung, colorectal, prostate, and other cancers. Colorectal cancer is one of the most commonly diagnosed cancers in the world that has recently been introduced as the third leading cause of cancer deaths in the world. microRNAs have a very crucial role in tumorgenesis and prevention of cancer, which plays a significant role with influencing various factors through different signaling pathways. Phosphoinositide 3 (PI3)‐kinase/AKT is one of the most important signaling pathways involved in the control and growth of tumor in colorectal cancer, through important proteins of this pathway, such as PTEN and AKT, that they can perform specific influence on this process. Our effort in this study is to collect microRNAs that act as tumor suppressors and oncomirs in this cancer through PI3‐kinase/AKT signaling pathway. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27415   open full text
• DDIT3 regulates cementoblast mineralization by isocitrate dehydrogenase 1 through nuclear factor‐κB pathway.
  Xiayi Liu, Hualing Sun, Miao Yu, Jie Liu, Beining Yang, Yanru Wu, Jiawei Wang.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- In this study, we reported the decreased expression of DDIT3 during cementogenic mineralization, as well as the inhibitory effect of DDIT3 to cementogenic mineralization. In addition, we showed that DDIT3 can regulate isocitrate dehydrogenase 1 through the nuclear factor‐κB pathway. Abstract DDIT3 is of great importance in endoplasmic reticulum stress and is involved in many inflammatory diseases and mineralization processes. The cementum protects teeth from periodontitis and provides attachment for Sharpey's fibers of the periodontal ligament. However, the effect of DDIT3 on cementoblast differentiation remains largely unknown. In this study, we found that DDIT3 was suppressed during cementoblast differentiation. Knockdown of DDIT3 increased the messenger RNA (mRNA) and protein levels of several key osteogenic markers in vitro, including alkaline phosphatase, runt‐related transcription factor 2, and osteocalcin (OCN). In addition, isocitrate dehydrogenase 1 (IDH1) was increased during cementoblast differentiation, and knockdown of DDIT3 increased the protein and mRNA levels of IDH1. Furthermore, inhibition of IDH1 could partially reduce the effect of DDIT3 on cementoblast differentiation. The DDIT3 knockdown activated nuclear factor‐κB (NF‐κB) transcriptional activity and upregulated the expression of p‐p65 and p‐IκBα. The increased osteogenic differentiation ability and IDH1 expression, as induced by the DDIT3 knockdown, could be partially turned over by the addition of NF‐κB inhibitor BAY 11–7082. Overall, our data clarified that DDIT3 suppresses cementoblast differentiation through IDH1, via the NF‐κB pathway. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27811   open full text
• Ranolazine prevents pressure overload‐induced cardiac hypertrophy and heart failure by restoring aberrant Na+ and Ca2+ handling.
  Jiali Nie, Quanlu Duan, Mengying He, Xianqing Li, Bei Wang, Chi Zhou, Lujin Wu, Zheng Wen, Chen Chen, Dao Wu Wang, Katherina M. Alsina, Xander H.T. Wehrens, Dao Wen Wang, Li Ni.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2 In this study, we revealed the effect of ranolazine to attenuate pressure overload‐induced cardiac hypertrophy and heart failure in mice by reducing Na+ accumulation and Ca 2+ overload, normalizing SR leak. The Ca 2+‐dependent Ca 2+/calmodulin (CaM)/CaMKII/MEF2D and CaM/CaMKII/calcineurin/nuclear factor of activated T‐cells hypertrophic pathways and endoplasmic reticulum (ER) stress were triggered by pressure overload and were inhibited by ranolazine. Abstract Background Cardiac hypertrophy and heart failure are characterized by increased late sodium current and abnormal Ca2+ handling. Ranolazine, a selective inhibitor of the late sodium current, can reduce sodium accumulation and Ca 2+ overload. In this study, we investigated the effects of ranolazine on pressure overload‐induced cardiac hypertrophy and heart failure in mice. Methods and Results Inhibition of late sodium current with the selective inhibitor ranolazine suppressed cardiac hypertrophy and fibrosis and improved heart function assessed by echocardiography, hemodynamics, and histological analysis in mice exposed to chronic pressure overload induced by transverse aortic constriction (TAC). Ca2+ imaging of ventricular myocytes from TAC mice revealed both abnormal SR Ca 2+ release and increased SR Ca 2+ leak. Ranolazine restored aberrant SR Ca 2+ handling induced by pressure overload. Ranolazine also suppressed Na + overload induced in the failing heart, and restored Na +‐induced Ca 2+ overload in an sodium‐calcium exchanger (NCX)‐dependent manner. Ranolazine suppressed the Ca 2+‐dependent calmodulin (CaM)/CaMKII/myocyte enhancer factor‐2 (MEF2) and CaM/CaMKII/calcineurin/nuclear factor of activated T‐cells (NFAT) hypertrophy signaling pathways triggered by pressure overload. Pressure overload also prolonged endoplasmic reticulum (ER) stress leading to ER‐initiated apoptosis, while inhibition of late sodium current or NCX relieved ER stress and ER‐initiated cardiomyocyte apoptosis. Conclusions Our study demonstrates that inhibition of late sodium current with ranolazine improves pressure overload‐induced cardiac hypertrophy and systolic and diastolic function by restoring Na+ and Ca 2+ handling, inhibiting the downstream hypertrophic pathways and ER stress. Inhibition of late sodium current may provide a new treatment strategy for cardiac hypertrophy and heart failure. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27791   open full text
• Effect of PLK1 inhibition on cisplatin‐resistant gastric cancer cells.
  Zihao Chen, Yanling Chai, Ting Zhao, Ping Li, Lihua Zhao, Fang He, Yu Lang, Jing Qin, Hongping Ju.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2 This study aims to investigate the effect of polo‐like kinase 1 (PLK1) inhibition on cisplatin (DDP)‐resistant gastric cancer (GC) cells. Together, our experimental results illustrated that the DDP resistance of GC cells might be associated with the aberrant overexpression of PLK1. PLK1 inhibition, including si‐PLK1 and BI2536 treatment, could restore the chemosensitivity of drug‐resistant SGC‐7901/DDP cells and enhance the efficacy of DDP, revealing the potential value of PLK1 inhibition in GC chemotherapy. Abstract Objective This study aims to investigate the effect of polo‐like kinase 1 (PLK1) inhibition on cisplatin (DDP)‐resistant gastric cancer (GC) cells. Methods: The transcriptional level of PLK1 was measured by quantitative reverse‐transcription polymerase chain reaction. Expressions of PLK1 and its downstream mediators as well as autophagy‐related protein LC3 I/LC3 II were detected by western blot. An 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and 5‐ethynyl‐2′‐deoxyuridine immunofluorescent staining were conducted to evaluate the cell viability and replication activity separately. Flow cytometry was carried out to determine the cell cycle status. The GFP‐LC3 vector contributed toward tracking the formation and aggregation of autophagosomes. Results: Drug‐resistant SGC‐7901/DDP cells showed insignificant changes in all phases after DDP treatment, including DNA replication, cell proliferation, cell cycle, and apoptosis, whereas DDP could significantly improve the autophagy level of SGC‐7901/DDP as well as PLK1expression. By downregulating the expression of PLK1, both BI2536 andsi‐PLK1 enhanced SGC‐7901/DDP sensitivity to DDP, suppressing the proliferation and autophagy as well as improving the apoptosis rate. PLK1 inhibition also resulted in the repression of cell division regulators CDC25C and cyclin B1. Conclusion: Together, our experimental results illustrated that the DDP resistance of GC cells might be associated with the aberrant overexpression of PLK1. PLK1 inhibition, including si‐PLK1 and BI2536 treatment, could restore the chemosensitivity of drug‐resistant SGC‐7901/DDP cells and enhance the efficacy of DDP, revealing the potential value of PLK1 inhibition in GC chemotherapy. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.26777   open full text
• Novel effects of sphingosylphosphorylcholine on the apoptosis of breast cancer via autophagy/AKT/p38 and JNK signaling.
  Di Ge, Jia Gao, Lina Han, Ying Li, Hong‐Hong Liu, Wan‐Cheng Yang, Fen Chang, Jing Liu, Mei Yu, Jing Zhao.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- Sphingosylphosphorylcholine (SPC) could induce apoptosis through c‐Jun N‐terminal kinase (JNK) signaling in MDA‐MB‐231 cells. An antagonistic signaling about autophagy/AKT/p38 was activated to inhibit JNK signaling and subsequent apoptosis by SPC. Though evoking autophagy/AKT/p38 pathways that anatagonize apoptosis, SPC eventually led to cell apoptotic death. Abstract Sphingosylphosphorylcholine (SPC), an important lipid mediator in blood, inhibits the proliferation and migration of various cancer cells. However, its effect as a cell‐specific sphingolipid in breast cancer cells is still unknown. Here, we showed that SPC promoted autophagy and apoptosis in triple‐negative breast cancer MDA‐MB‐231 cells. Autophagy worked as a negative regulator of apoptosis‐induced by SPC. Mechanistically, SPC mediated apoptosis via activating c‐Jun N‐terminal kinase (JNK). Meanwhile, p38MAPK (p38) and protein kinase B (PKB or AKT) signaling pathways were also activated to inhibit apoptosis, suggesting that SPC could evoke multiple signaling pathways to modulate cell apoptosis. In addition, the crosstalk between autophagy, p38, AKT and JNK is that autophagy, p38, and AKT attenuated the JNK. AKT and p38 were in the downstream of autophagy, which is autophagy/AKT/p38 signaling evoked by SPC to antagonize JNK signaling and subsequent apoptosis. Although the pathways that antagonize apoptosis were evoked, the cells eventually reached apoptosis by SPC. Therefore, the combination with pharmacological autophagy inhibitors would be a more effective therapeutic strategy for eliminating breast cancer cells by SPC. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27802   open full text
• Protective effect of microRNA‐224 on acute lower extremity ischemia through activation of the mTOR signaling pathway via CHOP in mice.
  Yang‐Xi Chen.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- MicroRNA‐224 could alleviate the occurrence and development of acute lower extremity ischemia (ALEXI) in mice through activation of the mammalian target of rapamycin (mTOR) signaling pathway by downregulating C/EBP homologous protein (CHOP). Abstract Acute lower extremity ischemia (ALEXI) is known worldwide as an urgent condition, occurring when there is an abrupt interruption in blood flow into an extremity. This study aims to investigate whether microRNA‐224 (miR‐224) affects the ALEXI mice and the underlying mechanism. The miR‐224 expression and C/EBP homologous protein (CHOP), mammalian target of rapamycin (mTOR), translation initiation factor 4E‐binding protein 1 (4E‐BP1), and phosphoprotein 70 ribosomal protein S6 kinase (p70S6K) messenger RNA (mRNA), as well as protein expressions, were determined. The target gene of miR‐224 was also verified by using a luciferase reporter gene assay. The vascular endothelial cells from the ALEXI mice were transfected with miR‐224 mimics, miR‐224 inhibitors, or small‐interfering RNA against CHOP. Cell proliferation was assessed using a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. The cell cycle distribution along with the cell apoptosis were both evaluated by using a flow cytometry. The muscle fibers of the lower extremities found in the ALEXI mice were evidently swollen and rounded, presenting with a remarkably narrowed gap. The positive CHOP expression increased in ALEXI mice than normal mice, while the miR‐224 expression and mTOR, 4E‐BP1, and p70S6K mRNA, as well as the protein expression, decreased. Luciferase reporter gene assay validated that the miR‐224 gene directly targeted CHOP. MiR‐224 facilitated cell proliferation but inhibited cell apoptosis; by contrast, CHOP increased cell apoptosis. Moreover, the cells transfected along with miR‐224 mimic exhibited a lower CHOP expression as well as increased mTOR, 4E‐BP1, and p70S6K expression. Our study provided evidence that miR‐224 could alleviate the occurrence and development of ALEXI in mice through activation of the mTOR signaling pathway by downregulating CHOP. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27550   open full text
• Biogenic nanoselenium particles activate Nrf2‐ARE pathway by phosphorylating p38, ERK1/2, and AKT on IPEC‐J2 cells.
  Xiao Xiao, Deguang Song, Yuanzhi Cheng, Yuhan Hu, Fengqin Wang, Zeqing Lu, Yizhen Wang.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- Our data demonstrated that biogenic nanoselenium (BNS) particles activated Nrf2‐ARE pathway through p38, extracellular signal‐regulated kinase 1/2 (ERK1/2), and protein kinase B (AKT) mediated‐phosphorylation of nuclear factor (erythroid‐derived‐2)‐like 2 (Nrf2) to improve the antioxidant function of intestinal epithelial cells. Abstract As the intestinal epithelium is vulnerable to oxidative stress because of frequent enterocyte renewal and continuous exposure to exogenous agents, it is meaningful to figure out how the epithelial cells exert antioxidant function. We previously synthesized a novel biogenic nanoselenium (BNS) particles and proved that BNS could effectively improve intestinal antioxidative function through activating Nrf2‐ARE pathway. The objective of the present study was to investigate the mechanism by which BNS activate Nrf2‐ARE pathway on the physiological function of intestinal epithelial cells. In the present study, we demonstrated that treatment of IPEC‐J2 cells with BNS particles not only elevated the levels of downstream proteins of nuclear factor (erythroid‐derived‐2)‐like 2 (Nrf2) such as heme oxygenase‐1 and NQO‐1 in a time‐dependent manner which started to weaken at 12 hr after treatment but also significantly activated Nrf2, mitogen‐activated protein kinase (MAPK), and protein kinase B (AKT) pathway in a time‐dependent manner within 24 hr. BNS particles significantly increased the content of phosphorylated‐Nrf2, without evident influence on the level of Kelch‐like ECH‐associated protein 1 (Keap1). Moreover, BNS also induced the activation of p38, extracellular signal‐regulated kinase 1/2 (ERK1/2), c‐Jun N‐terminal kinase, and AKT while phosphorylating Nrf2. Using specific protein kinase inhibitors, we found that the Nrf2‐phosphorylating and antioxidative effects of BNS particles were abolished when p38, ERK1/2, and AKT were significantly inhibited. Overall, our data demonstrated that BNS particles activated Nrf2‐ARE pathway through p38, ERK1/2, and AKT mediated‐phosphorylation of Nrf2 to improve the antioxidant function of intestinal epithelial cells - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27773   open full text
• Prospect of mesenchymal stem cells in therapy of osteoporosis: A review.
  Leili Aghebati‐Maleki, Sanam Dolati, Reza Zandi, Ali Fotouhi, Majid Ahmadi, Ali Aghebati, Mohammad Nouri, Seyed Kazem Shakouri, Mehdi Yousefi.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- Stem cell therapy has been recommended as a probable therapeutic approach for patients with osteoporosis. Even though the concept of stem cell‐based therapy for osteoporosis has suggested substantial attention over the years, no clinical trial has been published in humans. The cell studies on the basis of osteoporosis are primarily attentive on osteoclastic activity and bone resorption procedures; at that point on osteoblastogenesis, and in recent times on the differentiation probable of mesenchymal stem cells. In this review, we have summarized the therapeutic role of stem cell‐based strategy in osteoporosis. Abstract Osteoporosis is a systemic skeletal disease associated with reduced bone strong point that results in raised fracture risk, with decreased bone strength, leading to reduced bone mineral density and poor bone quality. It is the most common in older females but some men are also at high risk. Although considered as a predictable result of aging, it is can be avoidable and treatable. The existing treatment of osteoporosis mainly contains antiresorptive and anabolic agents. In spite of these improvements, concerns around unusual side‐effects of antiresorptive drugs, and the lack of perfect confirmation in maintenance of their long‐standing effectiveness is bring about many patients not receiving these drugs. Over the years, the stem cell‐based therapy has attained substantial clinical consideration because of its potential to treat numerous diseases. The stem cell therapy has been recommended as a probable therapeutic approach for patients with osteoporosis. Even though the concept of stem cell‐based therapy for osteoporosis has caught substantial attention, no clinical trial has been published on humans. The cell studies based on osteoporosis are primarily focused on osteoclastic activity and bone resorption procedures. Earlier, it was on osteoblastogenesis and in recent times, on the differentiation probable of mesenchymal stem cells. In this review, we have summarized the therapeutic role of stem cell‐based strategy in osteoporosis. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27833   open full text
• Trehalose inhibits cell proliferation and amplifies long‐term temozolomide‐ and radiation‐induced cytotoxicity in melanoma cells: A role for autophagy and premature senescence.
  Giulia Allavena, Barbara Del Bello, Paolo Tini, Nila Volpi, Giuseppe Valacchi, Clelia Miracco, Luigi Pirtoli, Emilia Maellaro.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- Trehalose, a natural disaccharide, inhibits short‐term cell proliferation and, even more, colony‐forming capacity in melanoma cells. Moreover, trehalose magnifies temozolomide (TMZ)‐ and irradiation (IR)‐induced cytotoxicity in the long term. Two different cell responses are induced by trehalose: a remarkable autophagy in melanoma cells (A375) sensitive to TMZ‐ and IR‐induced apoptosis, and premature senescence in melanoma cells (SK‐Mel‐28) resistant to apoptosis and less prone to autophagy. Abstract Cutaneous melanomas frequently metastasize to the brain, with temozolomide (TMZ) plus radiotherapy (RT) offering little control of these lesions. We tested whether trehalose, a natural glucose disaccharide proved to induce autophagy, could enhance the effect of TMZ and ionizing radiation (IR). In two melanoma cell lines (A375 and SK‐Mel‐28), which greatly differ in chemosensitivity and radiosensitivity, trehalose significantly inhibited short‐term cell proliferation and also enhanced IR‐induced cytostasis. Interestingly, in TMZ‐resistant SK‐Mel‐28 cells, trehalose was more effective than TMZ, and combined trehalose + TMZ further reduced cell proliferation. In long‐term experiments, colony‐forming capacity was dramatically reduced by trehalose, and even more by combined trehalose + TMZ or trehalose + IR. In resistant SK‐Mel‐28 cells, although growth was inhibited most with trehalose + TMZ + IR‐6 Gy combined treatment, it is notable that trehalose + TMZ treatment was also very effective. Along with a direct antiproliferative effect, two further mechanisms may explain how trehalose potentiates TMZ‐ and IR‐induced effects: the remarkable trehalose‐stimulated autophagy in A375 cells, which were sensitive to TMZ‐ and IR‐induced apoptosis; and the notable trehalose‐stimulated premature senescence in SK‐Mel‐28 cells, which were resistant to apoptosis and less prone to autophagy. In normal melanocytes, trehalose induced a minor autophagy and cell proliferation inhibition, without affecting cell viability; moreover, when trehalose was used in combination with TMZ, the slight TMZ‐induced cytotoxicity was not significantly reinforced. Together, our results suggest that trehalose, a safe nutrient supplement able to cross the blood–brain barrier, is a promising candidate, worthy to be further explored in vivo, to augment the therapeutic efficacy of TMZ and RT in melanoma brain metastases. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27838   open full text
• Identification of dysregulated miRNAs in triple negative breast cancer: A meta‐analysis approach.
  Leimarembi Devi Naorem, Mathavan Muthaiyan, Amouda Venkatesan.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- The study adopted meta‐analysis using the robust rank aggregation method to integrate miRNA expression profiling datasets of triple negative breast cancer. A meta‐signatures of six significantly dysregulated miRNAs (hsa‐miR‐135b‐5p, hsa‐miR‐18a‐5p, hsa‐miR‐9‐5p, hsa‐miR‐522‐3p, hsa‐miR‐190b, and hsa‐miR‐449a) were identified from different studies and have high prediction accuracy. Abstract Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer with poor clinical outcomes and lack of approved targeted therapy. Dysregulated microRNAs (miRNAs) have been considered a promising biomarker, which plays an important role in the tumorigenesis of human cancer. Due to the increase in miRNA profiling datasets of TNBC, a proper analysis is required for studying. Therefore, this study used meta‐analysis to amalgamate ten miRNA profiling studies of TNBC. By the robust rank aggregation method, metasignatures of six miRNAs (4 upregulated: hsa‐miR‐135b‐5p, hsa‐miR‐18a‐5p, hsa‐miR‐9‐5p and hsa‐miR‐522‐3p; 2 downregulated: hsa‐miR‐190b and hsa‐miR‐449a) were obtained. The gene ontology analysis revealed that target genes regulated by miRNAs were associated with processes like the regulation of transcription, DNA dependent, and signal transduction. Also, it is noted from the pathway analysis that signaling and cancer pathways were associated with the progression of TNBC. A Naïve Bayes‐based classifier built with miRNA signatures discriminates TNBC and non‐TNBC samples in test data set with high diagnostic sensitivity and specificity. From the analysis carried out by the study, it is suggested that the identified miRNAs are of great importance in improving the diagnostics and therapeutics for TNBC. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27839   open full text
• LINC00707 contributes to hepatocellular carcinoma progression via sponging miR‐206 to increase CDK14.
  Janfei Tu, Zhongwei Zhao, Min Xu, Minjiang Chen, Qiaoyou Weng, Jiangmei Wang, Jiansong Ji.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- We observed that long intergenic nonprotein‐coding RNA 707 (LINC00707) was increased in hepatocellular carcinoma (HCC) cells and LINC00707 silence was able to greatly repress HCC progression, and the correlation between LINC00707 and mciroRNA‐206 (miR‐206) was validated in our research and downregulation of LINC00707 increased miR‐206 expression in HCC cells. Then, cyclin‐dependent kinase 14 (CDK14) was predicted as a target of miR‐206 and the correlation between them was proved in our study. In conclusion, our data implied that LINC00707/miR‐206/CDK14 axis participated in HCC development and LINC00707 might be a biomarker for HCC. Abstract Recently, increasing numbers of long noncoding RNAs (lncRNAs) have been found to be aberrantly expressed in various cancers. However, the roles of lncRNAs in hepatocellular carcinoma (HCC) progression is largely unknown. In our current study, we identified that long intergenic nonprotein‐coding RNA 707 (LINC00707) was remarkably elevated in HCC cells, indicating that LINC00707 was involved in HCC development. Subsequently, LINC00707 was significantly decreased in HepG2 and Huh7 cells. The in vitro functional assays demonstrated that knockdown of LINC00707 significantly reduced HCC cell proliferation, induced cell apoptosis, and blocked the cell cycle progression. In addition, HCC cell migration and invasion was also greatly inhibited by downregulation of LINC00707. Increasing evidence has indicated that lncRNAs can act as molecular sponges of microRNAs. Currently, we observed that microRNA‐206 (miR‐206) was dramatically inhibited in HCC cells and LINC00707 can modulate HCC development through sponging miR‐206. The binding correlation between LINC00707 and miR‐206 was confirmed by dual‐luciferase reporter assay, RNA pull down and RNA immunoprecipitation assay in our study. Moreover, cyclin‐dependent kinase 14 (CDK14) was predicted as a target of miR‐206 and we found that miR‐206 suppressed CDK14 levels in HCC cells. Finally, in vivo assays were used and it was proved that silence of LINC00707 can restrain HCC development through modulating miR‐206 to upregulate CDK14. In conclusion, it was implied that LINC00707 can lead to HCC progression through sponging miR‐206 and modulating CDK14. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27737   open full text
• Immunoresolvents in asthma and allergic diseases: Review and update.
  Ramin Lotfi, Alireza Rezaiemanesh, Seyed Hamidreza Mortazavi, Ali Gorgin Karaji, Farhad Salari.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- Asthma and allergic diseases are inflammatory conditions developed by excessive reaction of the immune system against normally harmless environmental substances. Essential fatty‐acids‐derived immunoresolvents, namely, lipoxins, resolvins, protectins, and maresins are anti‐inflammatory compounds that are believed to have protective and beneficial effects in inflammatory disorders, including asthma and allergies. Abstract Asthma and allergic diseases are inflammatory conditions developed by excessive reaction of the immune system against normally harmless environmental substances. Although acute inflammation is necessary to eradicate the damaging agents, shifting to chronic inflammation can be potentially detrimental. Essential fatty‐acids‐derived immunoresolvents, namely, lipoxins, resolvins, protectins, and maresins, are anti‐inflammatory compounds that are believed to have protective and beneficial effects in inflammatory disorders, including asthma and allergies. Accordingly, impaired biosynthesis and defective production of immunoresolvents could be involved in the development of chronic inflammation. In this review, recent evidence on the anti‐inflam]matory effects of immunoresolvents, their enzymatic biosynthesis routes, as well as their receptors are discussed. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27836   open full text
• Gene polymorphisms and serum levels of TL1A in patients with rheumatoid arthritis.
  Zhi‐Chao Yuan, Jia‐Min Wang, Lin‐Chong Su, Wang‐Dong Xu, An‐Fang Huang.
  Journal of Cellular Physiology. November 29, 2018
  --- - |2- 1. TNF‐like ligand 1A (TL1A) gene polymorphisms correlated with RA. 2. TL1A protein concentrations were elevated in RA patients. 3. TL1A may play important roles in RA. Abstract Recent findings showed elevated expression of tumor necrosis factor (TNF)‐like ligand 1A (TL1A) in rheumatoid arthritis (RA) patients and arthritis mice. However, whether TL1A gene polymorphisms may correlate with RA susceptibility needs to be discussed. This case‐control study was performed on 350 RA patients and 556 healthy subjects to identify TL1A genetic variants (rs3810936, rs6478109, and rs7848647) and their possible association with TL1A levels, susceptibility to and severity of RA. Odds ratio and 95% confidence interval were calculated to represent the correlation between TL1A polymorphisms and RA. The TL1A serum levels were evaluated. Results showed that frequencies of TC, TT + TC genotypes of rs3810936, rs7848647 in RA patients were significantly lower in RA patients compared with controls. Patients with C allele showed more severe disease course (disease activity index: erythrocyte sedimentation rate, rheumatoid factor) than in carriers of T allele. However, the allele or genotype frequencies of rs6478109 were not associated with RA. In addition, TL1A genetic variants conferred higher TL1A levels in RA patients compared with controls. In conclusion, these findings indicated an association between TL1A rs3810936, rs7848647 variation and the susceptibility of RA in a sample of Chinese individuals, and TL1A may correlate with severity of RA. - 'Journal of Cellular Physiology, EarlyView. '

  November 29, 2018   doi: 10.1002/jcp.27834   open full text
• The novel relationship between Sirt3 and autophagy in myocardial ischemia–reperfusion.
  Yitian Zheng, Binhao Shi, Mengqing Ma, Xiaoqin Wu, Xianhe Lin.
  Journal of Cellular Physiology. November 28, 2018
  --- - |2- The signal pathways in Sirt3‐mediated autophagy when myocaridal ischemia–reperfusion (I/R) happened. We tend to find the novel relationship between Sirt3 and autophagy when myocardial I/R happened. Abstract Class III histone deacetylases (HDACs) belong to the proteasome family, comprising seven family members identified in mammalian cells, identified Sirt1–Sirt7. As an important member of HDACs, Sirt3 is hotly debated for its multiple functions. It was reported that Sirt3 got involved in the alleviation of multiple diseases, including myocardial infarction, neuron ischemia, hypertrophy, and diabetic myopathy. Through regulating many cellular mechanisms, such as apoptosis, autophagy, and clearance of reactive oxygen species (ROS), Sirt3 played an important role in the alleviation of myocardial ischemia–reperfusion injury. Nowadays Sirt3‐induced autophagy was indicated to be involved in the process of the development of myocardial ischemia–reperfusion injury. Sirt3 could both activate and inhibit autophagy process by activating different downstream signal pathways, such as Sirt3–AMP‐activated protein kinase pathway, Sirt3–Foxo3a pathway, and Sirt3–superoxide dismutase–mitochondrial ROS pathway. Whereas the Sirt3‐induced autophagy in different phases of myocardial ischemia–reperfusion has not been systematically illustrated. In this review, we summarized the regulated mechanisms found in these years and listed the updated research about the relationship between Sirt3 and autophagy which are both positive and negative during myocardial ischemia–reperfusion phase. We anticipated that we may controlled the activation of autophagy by regulating the concentration of Sirt3 in myocyte. By maintaining a proper expression of autophagy in different phases of myocardial ischemia–reperfusion, we could reduce the morbidity of patients with myocardial infarction apparently in the future. - 'Journal of Cellular Physiology, EarlyView. '

  November 28, 2018   doi: 10.1002/jcp.27329   open full text
• EW‐7197 prevents ulcerative colitis‐associated fibrosis and inflammation.
  Maryam M. Binabaj, Fereshteh Asgharzadeh, Amir Avan, Farzad Rahmani, Atena Soleimani, Mohammad R. Parizadeh, Gordon A. Ferns, Mikhail Ryzhikov, Majid Khazaei, Seyed M. Hassanian.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Here, we investigate the potential therapeutic effects of EW‐7197 in a murine model of ulcerative colitis. Our results suggest that EW‐7197 has potentially useful therapeutic properties against colitis, with clinically translational potential of inhibiting key pathological responses of inflammation and fibrosis in patients with colitis. Abstract EW‐7197 is a transforming growth factor‐β type I receptor kinase inhibitor with potential anti‐inflammatory and antifibrotic properties. Here, we investigate the potential therapeutic effects of EW‐7197 in a murine model of ulcerative colitis. EW‐7197 attenuated the colitis disease activity index by improving rectal bleeding, body weight, and degree of stool consistency. EW‐7197 also reduced colorectal tissue damage and the colon histopathological score by reducing crypt loss, mucosal damage, and tissue inflammation. Moreover, EW‐7197 appeared to ameliorate the inflammatory and fibrotic responses by reducing oxidative stress, reducing submucosal edema and inflammatory cell infiltration, downregulating proinflammatory and pro‐fibrotic genes, and inhibiting excessive collagen deposition in inflamed and fibrotic ulcerative colitis tissues. These results suggest that EW‐7197 has potentially useful therapeutic properties against colitis, with clinically translational potential of inhibiting key pathological responses of inflammation and fibrosis in patients with colitis. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27823   open full text
• Poly (3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) improved osteogenic differentiation of the human induced pluripotent stem cells while considered as an artificial extracellular matrix.
  Fatemeh Sadat Hosseini, Fatemeh Soleimanifar, Amir Aidun, Seyedeh Elnaz Enderami, Ehsan Saburi, Hadi Zare Marzouni, Mohammad‐Mehdi Khani, Arash Khojasteh, Abdolreza Ardeshirylajimi.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- ‐The pluripotent stem cells produced from the patient and the nanofibrous polymeric scaffold that can be completely degraded in the body and its produced monomers could be also usable and are the best options for this implant. ‐In this study, electrospun PHBV nanofibers were fabricated and characterized and then osteogenic differentiation of the human iPSCs was investigated while cultured on PHBV scaffold. ‐MTT, alkaline phosphatase activity, calcium content, gene expression, and western blot evaluations were confirmed osteogenic differentiation of the human iPSCs increased significantly when grown on PHBV nanofibers. Abstract Cocell polymers can be the best implants for replacing bone defects in patients. The pluripotent stem cells produced from the patient and the nanofibrous polymeric scaffold that can be completely degraded in the body and its produced monomers could be also usable are the best options for this implant. In this study, electrospun poly (3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (PHBV) nanofibers were fabricated and characterized and then osteogenic differentiation of the human‐induced pluripotent stem cells (iPSCs) was investigated while cultured on PHBV scaffold. MTT results showed that cultured iPSCs on PHBV proliferation were increased compared to those cultured on tissue culture polystyrene (TCPS) as the control. Alkaline phosphatase (ALP) activity and calcium content were also significantly increased in iPSCs cultured on PHBV compared to the cultured on TCPS under osteogenic medium. Gene expression evaluation demonstrated that Runx2, collagen type I, ALP, osteonectin, and osteocalcin were upregulated in iPSCs cultured on PHBV scaffold in comparison with those cultured on TCPS for 2 weeks. Western blot analysis have shown that osteocalcin and osteopontin expression as two major osteogenic markers were increased in iPSCs cultured on PHBV scaffold. According to the results, nanofiber‐based PHBV has a promising potential to increase osteogenic differentiation of the stem cells and iPSCs‐PHBV as a cell‐co‐polymer construct demonstrated that has a great efficiency for use as a bone tissue engineered bioimplant. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27807   open full text
• Antibody–drug conjugates (ADCs) for cancer therapy: Strategies, challenges, and successes.
  Meghdad Abdollahpour‐Alitappeh, Majid Lotfinia, Tohid Gharibi, Jalal Mardaneh, Behrouz Farhadihosseinabadi, Pegah Larki, Babak Faghfourian, Koushan Sineh Sepehr, Kazem Abbaszadeh‐Goudarzi, Ghasem Abbaszadeh‐Goudarzi, Behrooz Johari, Mohammad Reza Zali, Nader Bagheri.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Third‐generation antibody–drug conjugates are more effective, showing lower levels of unconjugated monoclonal antibodies, more‐stable linkers, improved pharmacokinetic properties, and safety profiles. Abstract Targeted delivery of therapeutic molecules into cancer cells is considered as a promising strategy to tackle cancer. Antibody–drug conjugates (ADCs), in which a monoclonal antibody (mAb) is conjugated to biologically active drugs through chemical linkers, have emerged as a promising class of anticancer treatment agents, being one of the fastest growing fields in cancer therapy. The failure of early ADCs led researchers to explore strategies to develop more effective and improved ADCs with lower levels of unconjugated mAbs and more‐stable linkers between the drug and the antibody, which show improved pharmacokinetic properties, therapeutic indexes, and safety profiles. Such improvements resulted in the US Food and Drug Administration approvals of brentuximab vedotin, trastuzumab emtansine, and, more recently, inotuzumab ozogamicin. In addition, recent clinical outcomes have sparked additional interest, which leads to the dramatically increased number of ADCs in clinical development. The present review explores ADCs, their main characteristics, and new research developments, as well as discusses strategies for the selection of the most appropriate target antigens, mAbs, cytotoxic drugs, linkers, and conjugation chemistries. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27419   open full text
• CtIP promotes G2/M arrest in etoposide‐treated HCT116 cells in a p53‐independent manner.
  Hongyu Chen, Jin Shan, Dandan Chen, Ruoxi Wang, Wenjing Qi, Hailong Wang, Yueshuang Ke, Wenguang Liu, Xianlu Zeng.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- CtIP can modulate the etoposide sensitivity of HCT116 cells by promoting G2/M phase arrest, which mainly through the ATR‐Chk1‐CDC25C pathway rather than the p53‐p21/GADD45a pathway. Abstract Acquired resistance to cytotoxic antineoplastic agents is a major clinical challenge in tumor therapy; however, the mechanisms involved are still poorly understood. In this study, we show that knockdown of CtIP, a corepressor of CtBP, promotes cell proliferation and alleviates G2/M phase arrest in etoposide (Eto)‐treated HCT116 cells. Although the expression of p21 and growth arrest and DNA damage inducible α (GADD45a), which are important targets of p53, was downregulated in CtIP‐deficient HCT116 cells, p53 deletion did not affect G2/M arrest after Eto treatment. In addition, the phosphorylation levels of Ser317 and Ser345 in Chk1 and of Ser216 in CDC25C were lower in CtIP‐deficient HCT116 cells than in control cells after Eto treatment. Our results indicate that CtIP may enhance cell sensitivity to Eto by promoting G2/M phase arrest, mainly through the ATR‐Chk1‐CDC25C pathway rather than the p53‐p21/GADD45a pathway. The expression of CtIP may be a useful biomarker for predicting the drug sensitivity of colorectal cancer cells. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27824   open full text
• Long noncoding RNA LINC00460 promotes carcinogenesis via sponging miR‐613 in papillary thyroid carcinoma.
  Li Feng, Bin Yang, Xiao‐Di Tang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- The present study first demonstrated that the expression of long intergenic noncoding RNA 460 (LINC00460) was upregulated in papillary thyroid carcinoma (PTC) tissues and cell lines. And the elevation of LINC00460 was correlated with advanced tumor–node–metastasis (TNM) stage and lymph node metastasis. Our results also revealed that the oncogene LINC00460 promoted PTC progression via partly regulating miR‐613/sphingosine kinase 2 (SphK2) axis. These findings suggested that the LINC00460/miR‐613/SphK2 might act as a novel therapeutic target for the treatment of PTC. Abstract Long intergenic noncoding RNA 460 (LINC00460) has been identified as a critical regulator for multiple types of cancers. However, the biological role and underlying mechanism in human papillary thyroid carcinoma (PTC) still remain unclear and need to be uncovered. This study was aimed to ascertain the biological role and molecular mechanism of LINC00460 in PTC progression. Our findings revealed that the level of LINC00460 was significantly upregulated in PTC tissues and cell lines, which was positively correlated with advanced tumor–node–metastasis (TNM) stage and lymph node metastasis. Cellular experiments exhibited that knockdown of LINC00460 decreased proliferative, migratory, and invasive abilities of PTC cells. Mechanism assays noted that knockdown of LINC00460 suppressed cell proliferation, migration, and invasion, and inhibited expression of sphingosine kinase 2 (SphK2, a target of miR‐613) in PTC cells, at least in part, by regulating miR‐613. These findings suggested that LINC00460 could function as a competing endogenous RNA to regulate SphK2 expression by sponging miR‐613 in PTC. Targeting LINC00460 could be a promising therapeutic strategy for patients with PTC. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27799   open full text
• Intersectin‐Cdc42 interaction is required for orderly meiosis in porcine oocytes.
  Xiaoyan Li, Min Gao, Yongfu He, Bo Xiong, Honglin Liu, Ling Gu.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Our findings support a model where intersectins, through the interaction with Cdc42, modulates the assembly of meiotic apparatus and actin polymerization, consequently ensuring the orderly meiotic progression during porcine oocyte maturation. Abstract Intersectins (ITSNs) have been shown to act as adaptor proteins that govern multiple cellular events via regulating Cdc42 activity. However, it remains to be determined whether the ITSN‐Cdc42 pathway is functional in porcine oocytes. To address this question, we used a small molecule, ZCL278, to selectively disrupt the ITSN2‐Cdc42 interaction. In the present study, we find that porcine oocytes exposed to ZCL278 are unable to completely progress through meiosis. Meanwhile, the spindle defects and chromosomal congression failure are frequently detected in these oocytes. In support of this, we observed the accumulated distribution of vesicle‐like ITSN2 signals around the chromosome/spindle region during porcine oocyte maturation. In addition, our results also showed that inhibition of the ITSN‐Cdc42 interaction impairs the actin polymerization in porcine oocytes. In summary, the findings support a model where ITSNs, through the interaction with Cdc42, modulates the assembly of meiotic apparatus and actin polymerization, consequently ensuring the orderly meiotic progression during porcine oocyte maturation. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27510   open full text
• PCNA and JNK1‐Stat3 pathways respectively promotes and inhibits diabetes‐associated centrosome amplification by targeting at the ROCK1/14‐3‐3σ complex in human colon cancer HCT116 cells.
  Yu Cheng Lu, Pu Wang, Jie Wang, Ronald Ma, Shao Chin Lee.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- The study shows the power of functional proteomic analysis in the delineation of cell signal transduction. Understanding of the mechanisms underlying the diabetes‐associated centrosome amplification is beneficial for the development of intervention protocols for the control of centrosome amplification and its consequences (ie., cancer development) in diabetes. Abstract We have recently reported that type 2 diabetes promotes centrosome amplification via enhancing the expression, biding, and centrosome translocation of rho‐associated coiled‐coil containing protein kinase 1 (ROCK1)/14‐3‐3σ complex in HCT116 cells. In the functional proteomic study, we further investigated the molecular pathways underlying the centrosome amplification using HCT116 cells. We found that treatment of HCT116 cells with high glucose, insulin, and palmitic acid triggered the centrosome amplification and increased the expressions of proliferating cell nuclear antigen (PCNA), nucleophosmin (NPM), and 14‐3‐3σ. Individual knockdown of PCNA, NPM, or 14‐3‐3σ inhibited the centrosome amplification. Knockdown of PCNA inhibited the treatment‐increased expression of ROCK1, whereas knockdown of ROCK1 did not affect the PCNA expression. High glucose, insulin, and palmitic acid also increased the expressions of c‐Jun N‐terminal kinase‐1 (JNK1) and signal transducer and activator of transcription 3 (Stat3), individual knockdown of which upregulated the treatment‐increased expression of 14‐3‐3σ and promoted the centrosome amplification. In contrast, overexpression of JNK1 inhibited the centrosome amplification. Knockdown of Stat3 enhanced the centrosome translocation of 14‐3‐3σ. Moreover, we showed that knockdown of JNK1 inhibited the treatment‐increased expression of Stat3. Knockdown of PCNA, JNK1, or Stat3 did not have an effect on NPM and vice versa. In conclusion, our results suggest that PCNA and JNK1‐Stat3 pathways respectively promotes and feedback inhibits the centrosome amplification by targeting at the ROCK1/14‐3‐3σ complex, and NPM serves as an independent signal for the centrosome amplification. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27813   open full text
• CircRNA ZNF609 functions as a competitive endogenous RNA to regulate FOXP4 expression by sponging miR‐138‐5p in renal carcinoma.
  Yunhe Xiong, Jiabin Zhang, Chao Song.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Our research indicates that circRNA ZNF609 (circ‐ZNF609) functions as a competitive endogenous RNA to regulate forkhead box P4 expression by sponging microRNA‐138‐5p has great significance in the pathogenesis of renal carcinoma. Therefore, further study of circRNA may have clinical implications for future diagnosis and treatment of renal carcinoma and other diseases. Abstract Circular RNA (circRNA) play important roles in the pathological processes of many diseases. By analyzing the results of the GSE100186 chip, we found that the expression of circRNA ZNF609 (circ‐ZNF609) was significantly increased in renal cell carcinoma. Recently, there are studies showing that circ‐ZNF609 can regulate cell proliferation and invasion ability of various cells. In this study, we investigated whether circ‐ZNF609 may affect cell invasion and proliferation in renal carcinoma. Quantitative reverse transcription polymerase chain reaction was performed to detect the expression of circ‐ZNF609 in renal carcinoma cell lines and renal epithelial cells. The direct interaction between microRNA‐138‐5p (miR‐138‐5p) and forkhead box P4 (FOXP4) or circ‐ZNF609 was confirmed by luciferase reporter assay and RNA immunoprecipitation assay. We use Cell Counting Kit‐8, 5‐ethynyl‐2′‐deoxyuridine, and Matrigel assays to assess the effect of miR‐138‐5p or circ‐ZNF609 on cell proliferation or invasion ability. And we found that circ‐ZNF609 is significantly increased in renal carcinoma cell lines. In addition, the high expression of circ‐ZNF609 promotes cell proliferation and invasion ability. In short, our current study reveals the role of the circ‐ZNF609/miR‐138‐5p/FOXP4 regulatory network in renal carcinoma and provides a new perspective for the pathogenesis of renal carcinoma. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27744   open full text
• Active vitamin D regulates macrophage M1/M2 phenotypes via the STAT‐1‐TREM‐1 pathway in diabetic nephropathy.
  Xiaoliang Zhang, Yu Zhao, Xiaodong Zhu, Yinfeng Guo, Ying Yang, Yuteng Jiang, Bicheng Liu.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 Vitamin D can inhibit macrophage transition to the M1 phenotype through the STAT‐1/TREM‐1 pathway. Abstract Aim Imbalance of M1/M2 macrophages phenotype activation is a key point in diabetic nephropathy (DN). This study aimed to investigate whether active vitamin D (VD) suppresses macrophage transition to the M1 phenotype via inhibiting the high glucose‐induced STAT‐1 phosphorylation to reduce TREM‐1 expression. Methods In vivo, pathological changes in kidney tissue were detected and the expression of CD68 TREM‐1, STAT‐1, M1 makers, and M2 makers were acquired in renal tissue of patients with DN and 18w DN rats. In vitro, RAW 264.7 cells were incubated in the presence of high glucose with or without VD. Silencing and overexpression of TREM‐1 and silencing and activate of STAT‐1 were explored to elucidate the underlying mechanism. The expression of TREM‐1 and STAT‐1 and the changes of macrophage phenotype were examined separately by western blot and immunofluorescence staining. Results (a) Expression of TREM‐1, p‐STAT‐1, and M1 markers (iNOS and TNF‐α) were increased and positively correlated in kidneys from patients with DN. (b) In DN rats, the enlargement of glomerular surface area, expansion of glomerular mesangial matrix, the expression of CD68, TREM‐1, p‐STAT‐1, and M1 marker (iNOS) were significantly increased in comparison with the normal control group, whereas above changes were markedly decreased in the diabetic group treated with the VD group. (c) In vitro, VD significantly decreased high glucose‐induced CD68, TREM‐1, p‐STAT‐1, and M1 marker (iNOS) expression. However, above‐mentioned effects of VD are abolished when TREM‐1 is overexpressed or STAT‐1 is activated. Reductions in STAT‐1 expression decreased the TREM‐1 expression. Conclusion VD can inhibit macrophage transition to the M1 phenotype through the STAT‐1/TREM‐1 pathway. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27450   open full text
• Molecular interaction of NFκB and NICD in monocyte–macrophage differentiation is a target for intervention in atherosclerosis.
  Ambika Binesh, Sivasithamparam Niranjali Devaraj, Devaraj Halagowder.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Inhibition of NFκB regulated notch intracellular domain (NICD), which, in turn, downturned macrophage differentiation. Inhibition of both NFκB–NICD is a potential target for intervention in early stage of atherosclerosis (monocyte to macrophage differentiation), to prevent the furtherance of disease. Abstract The activation of two transcription factors, NFκB and NICD (notch intracellular domain), plays a crucial role in different stages of atherosclerotic disease progression, from early endothelial activation by modified lipids like oxidized low‐density lipoprotein (oxyLDL) to the imminent rupture of the atherosclerotic plaque. Inflammatory mediators and the notch pathway proteins were upregulated in atherogenic diet‐induced rats and the same was confirmed by the differentiation of monocyte to macrophage on exposure to oxyLDL. The inflammatory transcription factor NFκB and the notch signaling transcription factor NICD were analysed for their molecular interaction in monocyte to macrophage differentiation. Inhibition of NFκB by dexamethasone in monocyte to macrophage differentiation resulted in a concomitant downregulation of NICD, whereas inhibition of NICD by N‐(N‐[3, 5‐difluorophenacetyl])‐l‐alanyl)‐S‐phenylglycine t‐butyl ester (DAPT), a γ–secretase inhibitor, did not significantly influence the expression of NFκB, but downregulated macrophage differentiation. These findings revealed that NFκB inhibition using dexamethasone regulated NICD, which turned down macrophage differentiation. Thus, inhibition of both NFκB–NICD is a potential target for intervention in atherosclerosis. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27458   open full text
• G protein‐coupled receptor 119 is involved in RANKL‐induced osteoclast differentiation and fusion.
  Hyun‐Ju Kim, Hye‐Jin Yoon, Ji‐Wan Park, Xiangguo Che, Xian Jin, Je‐Yong Choi.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- G protein‐coupled receptor 119 (GPR119) activation inhibits osteoclastogenesis and fusion induced by receptor activator of nuclear factor‐κB ligand (RANKL). GPR119 may become a new target for the treatment of bone‐erosive diseases characterized by increased osteoclast number. Abstract G protein‐coupled receptor 119 (GPR119) is known to be a promising therapeutic target for type 2 diabetes. Recently, it has been reported that the GPR119 agonist increases bone mineral density in an animal model of diabetes, suggesting that GPR119 may play a key role in bone metabolism. In this study, we investigated the functional role of GPR119 in receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclast formation. We found that the GPR119 expression was markedly increased in preosteoclasts and then downregulated in mature osteoclasts. Activation of GPR119 with AS1269574, a potent selective agonist for GPR119, inhibited the generation of multinuclear osteoclasts from bone marrow‐derived macrophages. Confirming this observation, targeted silencing of GPR119 using short hairpin RNA abrogated the AS1269574‐mediated suppressive effect on osteoclast formation. GPR119 activation attenuated the expression of c‐Fos and nuclear factor of activated T cells cytoplasmic 1 (NFATc1) and blocked RANKL‐stimulated phosphorylation of IκBα, c‐Jun N‐terminal protein kinase (JNK), and extracellular signal‐regulated kinase (ERK) but not p38. In addition, GPR119 activation suppressed preosteoclast fusion by downregulating the expression of the dendritic cell‐specific transmembrane (DC‐STAMP), a molecule that is essential for cell–cell fusion in osteoclast formation. Furthermore, ectopic expression of DC‐STAMP restored AS1269574‐mediated inhibition of osteoclast fusion. Taken together, our findings demonstrate that GPR119 plays a negative role in osteoclast differentiation and fusion induced by RANKL, and therefore may represent a potential target for bone resorption‐associated diseases. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27805   open full text
• MicroRNA: A new player in response to therapy for colorectal cancer.
  Mostafa Sarvizadeh, Ziba V. Malekshahi, Ebrahim Razi, Hossein Sharifi, Nousin Moussavi, Mohsen Taghizadeh.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 Abstract Colorectal cancer (CRC) is one of the important malignancies that result in cancer‐related deaths worldwide. Multiple lines of evidence have indicated that different responses to therapy in CRC cells led to the failure of the current therapies. Hence, identification of the underlying cellular and molecular pathways involved in the emergence of different responses from CRC cells could contribute to finding and designing new therapeutic platforms to overcome the present limitations. Among the various targets involved in CRC pathogenesis, microRNAs (miRNAs) have key roles in many signaling pathways that are associated with the initiation and progression of CRC. Increasing evidence has confirmed that miRNAs as epigenetic regulators could play critical roles in the response (resistance or sensitivity) to therapy. Cancer stem cells are well‐known players in resistance to therapy in CRC. They have been shown to play significant roles via inhibition and activation of many miRNA networks. Hence, miRNAs could be involved in the resistance and sensitivity of therapy in CRC cells via affecting different mechanisms, such as activation of cancer stem cells. Here, we summarized the role of various miRNAs in response to therapy of CRC cells. Moreover, we highlighted the roles of these molecules in the function of cancer stem cells, which are known as important players in the resistance to therapy in CRC. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27806   open full text
• Downregulation of microRNA‐592 protects mice from hypoplastic heart and congenital heart disease by inhibition of the Notch signaling pathway through upregulating KCTD10.
  Xue‐Feng Pang, Xue Lin, Jian‐Jun Du, Ding‐Yin Zeng.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- This study demonstrates that downregulation of miR‐592 prevents CHD and hypoplastic heart by inhibition of the Notch signaling pathway via negatively binding to KCTD10. Abstract Evidence has demonstrated that the microRNA (miR) may play a significant role in the development of congenital heart disease (CHD). Here, we explore the mechanism of microRNA‐592 (miR‐592) in heart development and CHD with the involvement of KCTD10 and Notch signaling pathway in a CHD mouse model. Cardiac tissues were extracted from CHD and normal mice. Immunohistochemistry staining was performed to detect positive expression rate of KCTD10. A series of inhibitor, activators, and siRNAs was introduced to verified regulatory functions for miR‐592 governing KCTD10 in CHD. Furthermore, the effect of miR‐592 on cell proliferation and apoptosis was also investigated. Downregulated positive rate of KCTD10 was observed in CHD mice. Downregulation of miR‐592 would upregulate expression of KCTD10 and inhibit the activation of Notch signaling pathway, thus promote cell proliferation. This study demonstrates that downregulation of miR‐592 prevents CHD and hypoplastic heart by inhibition of the Notch signaling pathway via negatively binding to KCTD10. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27190   open full text
• α‐2‐Macroglobulin induces the shedding of microvesicles from cutaneous wound myofibroblasts.
  Alexandra Laberge, Akram Ayoub, Syrine Arif, Sébastien Larochelle, Alain Garnier, Véronique J. Moulin.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- α‐2‐Macroglobulin positively regulates microvesicle shedding through the activation of low‐density lipoprotein receptor‐related protein‐1 on myofibroblasts. Abstract Microvesicles (MVs) are recognized as an important class of cell‐to‐cell messengers. Although the properties of MVs are increasingly documented, the mechanisms regulating MV biogenesis remain debated. Myofibroblasts are a key cellular component of wound healing and have been shown to produce MVs upon stimulation with serum. However, the mediator(s) responsible for the observed effect of serum on MV release have yet to be identified. To isolate the molecule(s) of interest, serum proteins were sequentially separated using chromatography, selective precipitation, and electrophoresis. MV production was assessed throughout the purification and after stimulation of myofibroblasts with two potent purified molecules. α‐2‐Macroglobulin (A2M) was thereby found to dose‐dependently stimulate MV release. We confirmed the presence of the A2M receptor, low‐density lipoprotein receptor‐related protein‐1 (LRP1), on myofibroblasts. Inhibition of LRP1 resulted in a significant decrease in MV production. Together, our results suggest that A2M positively regulates MV shedding through the activation of LRP1 on myofibroblasts. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27794   open full text
• Central adiponectin induces trabecular bone mass partly through epigenetic downregulation of cannabinoid receptor CB1.
  Hua Jiang, Yuwei Wu, Paloma Valverde, Dana Murray, Jin Tang, Qi Yao, Qianqian Han, Jin Zhang, Lan Zhang, Lei Sui, Yin Tang, Qisheng Tu, Jake Chen.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- 1.Globular adiponectin (gAPN) enhanced expression levels of various histone deacetylases (HDACs), especially HDAC5. 2.Chromatin immunoprecipitation assays revealed HDAC5 bound to the transcriptional start site 2 (TSS2) region of the CB1 promoter. 3.Our study identified a possible novel central APN‐HDAC5‐CB1 signaling mechanism that promotes peripheral bone formation through epigenetic regulation of hypothalamic CB1 expression. Abstract Central adiponectin (APN) in either the globular (gAPN) or full‐length forms decreases sympathetic tone and increases trabecular bone mass in mice through the hypothalamus. It is known that cannabinoid type‐1 (CB1) receptors are expressed in the hypothalamic ventromedial nucleus and participate in energy metabolism by controlling sympathetic activity. However, whether central APN could influence endocannabinoid signaling through CB1 receptor to regulate bone metabolism has not been characterized. Here we demonstrate that gAPN downregulated CB1 expression in embryonic mouse hypothalamus N1 cells in vitro. gAPN intracerebroventricular (icv) infusions also decreased hypothalamic CB1 expression and bone formation parameters in APN‐knockout (APN‐KO) and wild‐type mice. Most importantly, mice pretreated with icv infusions with the CB1 receptor agonist arachidonyl‐2′‐chloroethylamine or antagonist rimonabant attenuated or enhanced respectively central APN induction of bone formation. We then investigated whether epigenetic signaling mechanisms were involved in the downregulation of hypothalamic CB1 expression by gAPN. We found gAPN enhanced expression levels of various histone deacetylases (HDACs), especially HDAC5. Furthermore, chromatin immunoprecipitation assays revealed HDAC5 bound to the transcriptional start site transcription start site 2 region of the CB1 promoter. In summary, our study identified a possible novel central APN‐HDAC5‐CB1 signaling mechanism that promotes peripheral bone formation through epigenetic regulation of hypothalamic CB1 expression. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27460   open full text
• A novel infram deletion in MSH6 gene in glioma: Conversation on MSH6 mutations in brain tumors.
  Zeinab Deris Zayeri, Maryam Tahmasebi Birgani, Javad Mohammadi Asl, Davood Kashipazha, Mohammadreza Hajjari.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 1. Making a connection between the genetic profiles and the evolved pathways and therapeutic approaches enhance our vision in estimating the results beyond the treatment. 2. our patients revealed a homozygous pattern for a novel 9 base‐pare deletion at the 912–914 codon on exon 4 of the MSH6 gene. 3. In temozolomide (TMZ)‐resitance cases, it is suggested to use complementary strategies Abstract Objective and background Histological and molecular information and biopsy help in the diagnosis of the type and grade of tumors and increase the value of estimation of the biological behavior of tumors. In this study, we focused on a consanguineous Iranian Family with high prevalence of brain tumors in their pedigree and reviewed the literature on MSH6 mutations in brain tumors and the treatment responses focused on Gliomas. Method We chose a family with a high prevalence of brain tumor in their pedigree. We studied the proband's neuroimaging and brain proton magnetic resonance spectroscopy (MRS), magnetic resonance imaging (MRI), biopsy result, and whole‐genome sequencing. Result The neuroimaging and brain proton MRS reported a lesion in the right frontoparietal. The MRI revealed a large enhancible heterogenous mass in the right temporo‐fronto‐parieto‐occipital lobes with involvement of corpus callosum which was suggestive of glioma. The patient revealed a homozygous pattern for a novel 9 base‐pare deletion at the 912–914 codon on exon 4 of the MSH6 gene. Discussion We discuss several studies on MSH6 mutations in brain tumors and we discuss treatment responses in MSH6 mutations and the studies conducted to sensitize chemotherapy and radiotherapy resistance brain tumors to face this subject efficiently. Conclusion Patients should be evaluated for MMR mutation before chemo and radiotherapy, and it is valuable to follow‐up these mutations during the treatment too. In temozolomide (TMZ)‐resitance cases, it is suggested to use complementary strategies such as using HDACis and a combination of a STAT3 Inhibitor and an mTOR inhibitor, BER inhibition mechanism, and PARP‐1 inhibitor. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27759   open full text
• Identification of functional lncRNAs in atrial fibrillation by integrative analysis of the lncRNA‐mRNA network based on competing endogenous RNAs hypothesis.
  Cheng Qian, Hang Li, Danqi Chang, Baozhu Wei, Yanggan Wang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- In this study, we identified eight functional long noncoding RNAs (lncRNAs) for atrial fibrillation based on competing endogenous RNA (ceRNA) hypothesis. These eight lncRNAs showed a good diagnostic power for atrial fibrillation (AF) and could be used as potential biomarkers Abstract A mounting body of evidence has suggested that long noncoding RNAs (lncRNAs) play critical roles in human diseases by acting as competing endogenous RNAs (ceRNAs). However, the functions and ceRNA mechanisms of lncRNAs in atrial fibrillation (AF) remain to date unclear. In this study, we constructed an AF‐related lncRNA‐mRNA network (AFLMN) based on ceRNA theory, by integrating probe reannotation pipeline and microRNA (miRNA)‐target regulatory interactions. Two lncRNAs with central topological properties in the AFLMN were first obtained. By using bidirectional hierarchical clustering, we identified two modules containing four lncRNAs, which were significantly enriched in many known pathways of AF. To elucidate the ceRNA interactions in certain disease or normal condition, the dysregulated lncRNA‐mRNA crosstalks in AF were further analyzed, and six hub lncRNAs were obtained from the network. Furthermore, random walk analysis of the AFLMN suggested that lncRNA RP11‐296O14.3 may function importantly in the pathological process of AF. All these eight lncRNAs that were identified from previous steps (RP11‐363E7.4, GAS5, RP11‐410L14.2, HAGLR, RP11‐421L21.3, RP11‐111K18.2, HOTAIRM1, and RP11‐296O14.3) exhibited a strong diagnostic power for AF. The results of our study provide new insights into the functional roles and regulatory mechanisms of lncRNAs in AF, and facilitate the discovery of novel diagnostic biomarkers or therapeutic targets. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27819   open full text
• Cardiovascular complications and related risk factors underlying opium consumption.
  Reyhaneh T. Ebdali, Seyyedeh Samaneh Tabaee, Samira Tabaei.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Opium might have deleterious influences on cardiovascular events. As a result, people need to be trained with regard to the adverse effects of opium on cardiovascular events. It may cause hypertension, dyslipidemia, and hormonal abnormalities. Abstract Opium is considered as the second most abused addictive compound in worldwide. It seems that one of the causes for common consumption of opium in many countries is a traditional belief, even among medical personnel, through which opium might have advantageous influences on cardiovascular events and be beneficial in controlling hypertension, dyslipidemia, and diabetes. According to several investigations, it is thought that opium not only has no beneficial effects on cardiovascular events, but it might have deleterious influences on these settings. As a result, people need to be trained with regard to the adverse effects of opium on cardiovascular events. In this review, we try to go through the understanding of the effects of opium cardiovascular disorders and related complications such as blood pressure, blood sugar, lipid circumstances, and finally atherosclerosis. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27780   open full text
• Inhibition of miR‐124 improves neonatal necrotizing enterocolitis via an MYPT1 and TLR9 signal regulation mechanism.
  Yiyu Yin, Zhenfang Qin, Xiaobing Xu, Xu Liu, Huaxin Zou, Xiaole Wu, Junhua Cao.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 miR‐124 was a promoter of necrotizing enterocolitis (NEC), which promotes the intestine cell apoptosis and inflammatory cell infiltration through the inhibition of Toll‐like receptor 9 (TLR9) expression by targeting rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1). Abstract Backgrounds Necrotizing enterocolitis (NEC) was one of the main causes of morbidity and mortality in neonates. Our objective was to detect the mechanism of miR‐124 in small bowel tissues of NEC. Methods Hematoxylin and eosin (H&E) staining was used to detect the repair of the damaged tissues in rat NEC model. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to evaluate the cell apoptosis level in intestinal tissue. Reverse transcription polymerase chain reaction (RT‐PCR) was used to detect the messenger RNA (mRNA) expression level of miR‐124, Rho‐associated coiled‐coil‐containing protein kinase 1 (ROCK1), myosin phosphatase target subunit 1 (MYPT1), and Toll‐like receptor 9 (TLR9) in NEC tissues and IEC‐6 cells. Luciferase reporter assay was used to verify whether ROCK1 is a direct target of miR‐124. Results miR‐124 was overexpressed in the NEC tissues, while ROCK1 and MYPT1 was downregulated in the NEC tissues. Inhibition of miR‐124, suppressed the intestinal cell apoptosis and promoted the expression of ROCK1 and MYPT1. What is more, overexpression of miR‐124 could inhibit the expression of ROCK1, TLR9, and MYPT1. Luciferase assay confirmed that miR‐124 can regulate the transcriptional activity of ROCK1 through binding its 3′‐UTR region. Conclusion miR‐124 was a promoter of NEC, which promotes the intestine cell apoptosis and inflammatory cell infiltration through the inhibition of TLR9 expression by targeting ROCK1. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27691   open full text
• MicroRNA‐99b suppresses human cervical cancer cell activity by inhibiting the PI3K/AKT/mTOR signaling pathway.
  Yong‐Jie Li, Yue Wang, Yi‐Ying Wang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- These results indicate that miR‐99b attenuates the migration and invasion of human cervical cancer cells through downregulation of the phosphatidylinositol 3‐kinase/protein kinase B/mechanistic target of rapamycin signaling pathway, which provides a therapeutic approach for cervical cancer treatment. Abstract Cervical cancer is common cancer among women with high morbidity. MicroRNAs (miRs) are involved in the progression and development of cervical cancer. This study aimed to explore the effect of miR‐99b‐5p (miR‐99b) on invasion and migration in cervical cancer through the phosphatidylinositol 3‐kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) signaling pathway. The microarray‐based analysis was used to screen out differentially expressed miRNAs. Expression of miR‐99b, PI3K, AKT, mTOR, and ribosomal protein S6 kinase (p70S6K) was determined in both cervical cancer tissues and paracancerous tissues. Next, alteration of miR‐99b expression in cervical cancer was conducted to evaluate levels of PI3K, AKT, mTOR, p70S6K matrix metallopeptidase 2, epithelial cell adhesion molecule, and intercellular adhesion molecule 1, as well as the effect of miR‐99b on cell proliferation, invasion, migration, cell cycle distribution, and apoptosis. The results demonstrated that miR‐99b expression was decreased and levels of PI3K, AKT, mTOR, and p70S6K were elevated in cervical cancer tissues. More important, overexpressed miR‐99b repressed the PI3K/AKT/mTOR signaling pathway, inhibited cell proliferation, invasion, and migration, blocked cell cycle entry, and promoted apoptosis in cervical cancer. These results indicate that miR‐99b attenuates the migration and invasion of human cervical cancer cells through downregulation of the PI3K/AKT/mTOR signaling pathway, which provides a therapeutic approach for cervical cancer treatment. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27645   open full text
• Altered plasma proteins released from platelets and endothelial cells are associated with human patent ductus arteriosus.
  Hai‐Tao Hou, Xi‐Zhang, Jun Wang, Li‐Xin Liu, Jian‐Feng Zhang, Qin Yang, Guo‐Wei He.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Ductus arteriosus closure involves functional and structural remodeling, controlled by many factors and we for the first time demonstrate six key differential plasma proteins in human patent ductus arteriosus patients using proteomic technology and present a model to illustrate the constriction and closure of ductus arteriosus. Those proteins are closely related to platelet activation and coagulation cascades, complement mannan‐binding‐lectin, and other systemic signaling pathways. Our findings for the first time indicate that the differential proteins involved in different pathways may play key roles in the nonclosure of the ductus arteriosus in humans and may be developed as biomarkers for diagnosis. All those findings may be served as the basis of understanding the etiology and pathogenesis of patent ductus arteriosus. Abstract Patent ductus arteriosus is the third most common congenital heart disease and resulted from the persistence of ductal patency after birth. Ductus arteriosus closure involves functional and structural remodeling, controlled by many factors. The changes in plasma protein levels associated with PDA closure are not known. Here we for the first time demonstrate six key differential plasma proteins in human patent ductus arteriosus patients using proteomic technology and present a model to illustrate the constriction and closure of ductus arteriosus. Differentially expressed proteins were analyzed by using isobaric tags for relative and absolute quantification and validated by enzyme‐linked immunosorbent assay in new samples. The proteomic data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD008568. We found 74 upregulated and 98 downregulated proteins in the plasma of patients with PDA. Five decreased proteins (platelet factor 4, fibrinogen, von Willebrand factor, collagen, and mannose binding lectin‐associated serine protease‐2) and one increased protein (fibronectin) may increase the risk of patent ductus arteriosus. Those proteins are closely related to platelet activation and coagulation cascades, complement mannan‐binding‐lectin, and other systemic signaling pathways. Our findings for the first time indicate that the differential proteins involved in different pathways may play key roles in the nonclosure of the ductus arteriosus in humans and may be developed as biomarkers for diagnosis. All those findings may be served as the basis of understanding the etiology and pathogenesis of patent ductus arteriosus. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27433   open full text
• The possible role of bacteria, viruses, and parasites in initiation and exacerbation of irritable bowel syndrome.
  Aref Shariati, Fateme Fallah, Ali Pormohammad, Ali Taghipour, Hossein Safari, Alireza salami chirani, Sahar Sabour, Mahmood Alizadeh‐Sani, Taher Azimi.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Irritable bowel syndrome (IBS) is a prolonged and disabling functional gastrointestinal disorder with the incidence rate of 18% in the world. The pathophysiology of the IBS is hardly understood, whereas several possible mechanisms, such as visceral hypersensitivity, irregular gut motility, abnormal brain–gut relations, and the role of infectious agents, are implicated in initiation and development this syndrome. It appears that the main obligatory pathogens correspond with the IBS disease. Abstract Irritable bowel syndrome (IBS) is a prolonged and disabling functional gastrointestinal disorder with the incidence rate of 18% in the world. IBS could seriously affect lifetime of patients and cause high economic burden on the community. The pathophysiology of the IBS is hardly understood, whereas several possible mechanisms, such as visceral hypersensitivity, irregular gut motility, abnormal brain–gut relations, and the role of infectious agents, are implicated in initiation and development of this syndrome. Different studies demonstrated an alteration in B‐lymphocytes, mast cells (MC), T‐lymphocytes, and cytokine concentrations in intestinal mucosa or systemic circulation that are likely to contribute to the formation of the IBS. Therefore, IBS could be developed in those with genetic predisposition. Infections’ role in initiation and exacerbation of IBS has been investigated by quite several clinical studies; moreover, the possible role of some pathogens in development and exacerbation of this disease has been described. It appears that the main obligatory pathogens correspond with the IBS disease, Clostridium difficile, Escherichia coli, Mycobacterium avium subspecies paratuberculosis, Campylobacter concisus, Campylobacter jejuni, Chlamydia trachomatis, Helicobacter pylori, Pseudomonas aeruginosa, Salmonella spp, Shigella spp, and viruses, particularly noroviruses. A number of pathogenic parasites (Blastocystis, Dientamoeba fragilis, and Giardia lamblia) may also be involved in the progression and exacerbation of the disease. Based on the current knowledge, the current study concludes that the most common bacterial, viral, and parasitic pathogens may be involved in the development and progression of IBS. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27828   open full text
• Cell fate regulation by reticulon‐4 in human prostate cancers.
  Hu Zhao, Weipeng Su, Changyan Zhu, Tengyue Zeng, Shunliang Yang, Weizhen Wu, Dong Wang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Our results showed that reticulon‐4 (RTN4) was downregulated and targeted by miR‐148a‐3p, and inhibited prostate cancer cell proliferation by inducing cell cycle arrest and senescence, suggesting that RTN4 may be an important molecular event linked to prostate cancer. Abstract Reticulon‐4 (RTN4), a reticulon family protein localized in the endoplasmic reticulum, is reported to be involved in multiple physiological processes like neuroendocrine secretion and membrane trafficking in neuroendocrine cells. Previous studies have presented a great potential of RTN4 for the treatment of autoimmune‐mediated demyelinating diseases and spinal cord injury regeneration. While interaction with Bcl‐2 and Bcl‐2‐like family in apoptosis modulation implicated its possible role in various human cancers. However, the investigation of this gene in prostate cancer is mainly ignored. Here in our current study, we focused on its role in prostate cancer and found that RTN4 DNA copy numbers were higher in prostate cancer than normal prostate gland while its RNA and protein expressions were relatively lower. Chromosomal neighbor gene EML6 had similar expression patterns with RTN4 in prostate cancer tissues and cell lines, and further research found that they could be both targeted by miR‐148a‐3p. Lentivirus‐mediated RTN4 overexpression potently inhibited DU145 and LNCaP cells proliferation. Cell cycle was blocked in G2/M phase and significant cell senescence was observed in RTN4 overexpressed prostate cancer cells. Finally, interaction networks in the normal prostate gland and cancer tissues further revealed that RTN4 maybe phosphorylated by MAPKAPK2 and FYN at tyrosine 591 and serine 107, respectively. All these results implied that RTN4 might somehow participate in prostate tumor progression, and this elicits possibility to develop or identify selective agents targeting RTN4 for prostate cancer therapy. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27704   open full text
• Effects of nebulized N­‐acetylcystein on the expression of HMGB1 and RAGE in rats with hyperoxia‐­induced lung injury.
  Junying Qiao, Lixia Chen, Xianjie Huang, Feifei Guo.
  Journal of Cellular Physiology. November 27, 2018
  --- - "\nHigh mobility group box 1 (HMGB1) and receptor for advanced glycation end product (RAGE) were involved in the pathogenesis of hyperoxia‐induced lung injury, inhalation of N‐acetylcysteine might alleviate hyperoxia‐induced lung injury by regulating the expression of HMGB1 and RAGE. \n\n\n\n\n\n\nAbstract\n\nObjective\nTo investigate the role of high mobility group box 1 (HMGB1) and receptor for advanced glycation end product (RAGE) in the lungs of hyperoxia‐induced rats and the effect of N‐­acetlycystein (NAC).\n\n\nMethods\nA model of hyperoxic lung injury was established, rats in the NAC intervention, and control, hyperoxia group were given nebulized NAC aerosol, nebulized same volume of saline once a day for 7 consecutive days, respectively. Wet/dry (\nW/\nD) ratio of the lungs was determined to evaluate the edema of the lung tissues. Conventional hematoxylin–eosin (HE) staining was used to observe the pathological changes of lung tissues. Immunohistochemical staining was used to investigate the expression of HMGB1 and RAGE in the lung tissues. Quantitative reverse‐transcription polymerase chain reaction and western blot analysis were used to measured the changes in the messenger RNA (mRNA) and protein expression of HMGB1 and RAGE, respectively.\n\n\nResults\nWeight gain of the rats in the hyperoxia group was significantly slower than that in the control group and intervention group (p < 0.05). HE staining results showed lung tissues in the hyperoxia group were severely damaged compared with control group. W/D ratio in hyperoxia group was significantly higher than that in control group and intervention group (p < 0.05). Protein and mRNA expression of HMGB1 and RAGE in the hyperoxia group were significantly higher than control group and intervention group (p < 0.05).\n\n\nConclusion\nHMGB1 and RAGE were involved in the pathogenesis of hyperoxia‐induced lung injury, inhalation of NAC might alleviate hyperoxia‐induced lung injury by regulating the expression of HMGB1 and RAGE.\n" - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27724   open full text
• Cholinergic activity regulates the secretome of epicardial adipose tissue: Association with atrial fibrillation.
  Marinela Couselo‐Seijas, Jose N. López‐Canoa, Rosa M. Agra‐Bermejo, Esther Díaz‐Rodriguez, Angel L. Fernandez, Jose M. Martinez‐Cereijo, Dario Durán‐Muñoz, Susana B. Bravo, Alba Velo, Laila González‐Melchor, Xesus A. Fernández‐López, Jose L. Martínez‐Sande, Javier García‐Seara, Jose R. González‐Juanatey, Moises Rodriguez‐Mañero, Sonia Eiras.
  Journal of Cellular Physiology. November 27, 2018
  --- - "\n\n\nMuscarinic receptor type 3 is upregulated after adipogenesis.\nEpicardial fat from atrium or ventricles express this receptor and contain cholinergic activity.\nEpicardial fat‐released proteins after cholinergic activity as possible predictors of postoperative AF\n\n\n\n\n\nAbstract\nBotulinum toxin injection on epicardial fat, which inhibits acetylcholine (ACh) release, reduced the presence of atrial fibrillation (AF) in patients after heart surgery. Thus, we wanted to study the profile of the released proteins of epicardial adipose tissue (EAT) under cholinergic activity (ACh treatment) and their value as AF predictors. Biopsies, explants, or primary cultures were obtained from the EAT of 85 patients that underwent open heart surgery. The quantification of muscarinic receptors (mAChR) by real‐time polymerase chain reaction or western blot showed their expression in EAT. Moreover, mAChR Type 3 was upregulated after adipogenesis induction (p < 0.05). Cholinergic fibers in EAT were detected by vesicular ACh transporter levels and/or acetylcholinesterase activity. ACh treatment modified the released proteins by EAT, which were identified by nano‐high‐performance liquid chromatography and TripleTOF analysis. These differentially released proteins were involved in cell structure, inflammation, or detoxification. After testing the plasma levels of alpha‐defensin 3 (inflammation‐involved protein) of patients who underwent open heart surgery (\nn = 24), we observed differential levels between the patients who developed or did not develop postsurgery AF (1.58 ± 1.61 ng/ml vs. 6.2 ± 5.6 ng/ml; \np < 0.005). The cholinergic activity on EAT might suggest a new mechanism for studying the interplay among EAT, autonomic nervous system dysfunction, and AF." - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27723   open full text
• Knockdown of NEAT1 repressed the malignant progression of glioma through sponging miR‐107 and inhibiting CDK14.
  Yingwei Zhen, Yang Nan, Shewei Guo, Longzhou Zhang, Ge Li, Saichao Yue, Xianzhi Liu.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- We found that inhibition of nuclear paraspeckle assembly transcript 1 (NEAT1) repressed glioma development via sponging miR‐107. The correlation between NEAT1 and miR‐107 was validated in our study, and cyclin dependent kinase 14 (CDK14) was targeted by miR‐107 by using bioinformatics analysis. All these data revealed that NEAT1/miR‐107/CDK14 axis was involved in glioma development Abstract Aberrant expressions of long noncoding RNAs (lncRNAs) contribute to carcinogenesis via regulating tumor suppressors or oncogenes. LncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) has been recognized as an oncogene to promote tumor progression of many cancers. However, the function of NEAT1 in glioma remains poorly discovered. Currently, we focused on the role of NEAT1 in glioma. Here, we found that NEAT1 was greatly upregulated in glioma cells compared with normal human astrocytes (NHAs). Meanwhile, miR‐107 was significantly downregulated in glioma cell lines. Then, we observed that knockdown of NEAT1 suppressed the growth and invasion of glioma cells including U251 and SW1783 cells. Reversely, overexpression of NEAT1 dramatically induced glioma cell survival, increased cell colony formation, and promoted cell invasion ability. Subsequently, bioinformatics analysis was performed to predict the correlation between NEAT1 and miR‐107. Moreover, it was revealed that NEAT1 could modulate miR‐107 via serving as an endogenous sponge of miR‐107. The direct binding correlation between NEAT1 and miR‐107 was validated in our study. In addition, cyclin dependent kinase 14 (CDK14) was predicted as an messenger RNA target of miR‐107 and the association between them was confirmed in our research. Moreover, we implied that NEAT1 demonstrated its biological functions via regulating miR‐107 and CDK14 in vivo. In summary, our findings indicated that NEAT1/miR‐107/CDK14 axis participated in glioma development. NEAT1 could act as a significant prognostic biomarker in glioma progression. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27727   open full text
• c‐Met inhibition is required for the celecoxib‐attenuated stemness property of human colorectal cancer cells.
  Yueh‐Ming Lin, Chien‐Chang Lu, Yi‐Ping Hsiang, Shu‐Chuan Pi, Chih‐I Chen, Kung‐Chuan Cheng, Hsiao‐Lin Pan, Pei‐Hsuan Chien, Yun‐Ju Chen.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- This is the first report that elucidates the mechanism underlying celecoxib‐inhibited cancer stem cell (CSC). c‐Met is identified as a critical factor for the CSC property of colorectal cancer cells. Abstract Cyclooxygenase‐2 (COX‐2) is frequently overexpressed and enhances colorectal cancer (CRC) tumorigenesis, including cancer stem cell (CSC) regulation. Accordingly, nonsteroidal anti‐inflammatory drugs (NSAIDs), inhibiting COX‐1/2 activity, are viewed as potential drugs for CRC treatment. Accumulated evidence indicates that celecoxib has the most potency for antitumor growth among NSAIDs and the underlying mechanism is only partly dependent on COX‐2 inhibition. However, the potency of these NSAIDs on CSC inhibition is still not known. In this study, we found that among these NSAIDs, celecoxib has the most potency for CSC inhibition of CRC cells, largely correlating to inhibition of c‐Met, not COX‐2. Further analysis reveals that c‐Met activity was required for basal CSC property. Silence of c‐Met blocked whereas overexpression of c‐Met enhanced the celecoxib‐inhibited CSC property. Collectively, these results not only first elucidate the mechanism underlying celecoxib‐inhibited CSC but also indicate c‐Met as a critical factor for the CSC property of CRC cells. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27701   open full text
• Preparation of adriamycin gelatin microsphere‐loaded decellularized periosteum that is cytotoxic to human osteosarcoma cells.
  Chuan Chen, Jianghui Dong, Hong Chen, Xin Wang, Jin Mei, Liping Wang, Cory J Xian.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- The effectiveness of these two scaffolds in killing human osteosarcoma cells was tested by evaluating cell viability overtime of the cancer cells cultured with the scaffolds. In summary, a gelatin/decellularized periosteum‐based biologic scaffold material was designed aiming for local delivery of chemotherapy drugs for osteosarcoma, with the results showing ability of the scaffolds in sustaining release of the cancer drug and in suppressing growth of the cancer cells in vitro Abstract The purpose of this study was to develop a novel approach to treat bone osteosarcoma using a multipurpose scaffold aiming for local drug delivery. The slowly releasing microspheres was designed to deliver the chemotherapy drug adriamycin (ADM) and a decellularized (D) periosteum scaffold (which is known to be able to promote bone regeneration) was used to carry these microspheres. D‐periosteum was obtained by physical and chemical decellularization. Histological results showed that the cellular components were effectively removed. The D‐periosteum showed an excellent cytocompatibility and the ability to promote adhesion and growth of fibroblasts. Two kinds of slowly releasing microspheres, adriamycin gelatin microspheres (ADM‐GMS) and adriamycin poly (dl‐lactide‐co‐glycolide) gelatin microspheres (ADM‐PLGA‐GMS), were prepared and anchored to D‐periosteum, resulting in two types of drug‐releasing regenerative scaffolds. The effectiveness of these two scaffolds in killing human osteosarcoma cells was tested by evaluating cell viability overtime of the cancer cells cultured with the scaffolds. In summary, a gelatin/decellularized periosteum‐based biologic scaffold material was designed aiming for local delivery of chemotherapy drugs for osteosarcoma, with the results showing ability of the scaffolds in sustaining release of the cancer drug and in suppressing growth of the cancer cells in vitro. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27753   open full text
• Embryo quality, and not chromosome nondiploidy, affects mitochondrial DNA content in mouse blastocysts.
  Ying Jing, Li Li, Yuan‐Yuan Li, Ying‐Chun Ouyang, Qing‐Yuan Sun, Cui‐Lian Zhang, Rong Li.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Mitochondria prematurely replicate in blastocysts from aged women and in nondiploidy blastocysts, which have decreased developmental potential. Here, we show that the cytoplasmic quality, and not chromosome ploidy, causes premature mitochondrial biogenesis in early embryo development. Abstract It has been shown recently that there is premature mitochondria biosynthesis in blastocysts from older women whose egg or embryo quality is poor and that aneuploid blastocysts also have a high number of mitochondrial DNA (mtDNA) copies. Whether nondiploidy/aneuploidy or reduced egg or embryo quality causes premature mitochondrial biosynthesis is not known. This study constructed haploid, diploid, triploid, and tetraploid blastocysts by parthenogenetic activation, intracytoplasmic sperm injection with one or two sperm heads, blastomere electrofusion, respectively, and generated reduced cytoplasm quality embryos from diabetic mouse and in vitro fertilization of aged oocytes, and examined whether nondiploidy or reduced cytoplasm quality causes premature mitochondrial biosynthesis. MtDNA numbers of each blastocyst from different models were tested by absolute quantitative real‐time polymerase chain reaction. It was found that mtDNA content in preimplantation embryos was not associated with their chromosome ploidy, while mtDNA copy numbers in embryos with suboptimal quality were increased. Therefore, it might be the reduced cytoplasmic quality, and not chromosome nondiploidy, that causes premature mitochondria biosynthesis in blastocysts. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27713   open full text
• miR‐142‐3p as tumor suppressor miRNA in the regulation of tumorigenicity, invasion and migration of human breast cancer by targeting Bach‐1 expression.
  Behzad Mansoori, Ali Mohammadi, Mehri Ghasabi, Solmaz Shirjang, Razeieh Dehghan, Vahid Montazeri, Uffe Holmskov, Tohid Kazemi, Pascal Duijf, Morten Gjerstorff, Behzad Baradaran.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 Our results revealed that miR‐142‐3p could target Bach‐1in breast cancer cells leading to the reduction of EMT‐related proteins and reduced cell proliferation, invasion, and migration. The results also demonstrated that miR‐142‐3p could regulate important tumor suppressor miRNAs in breast cancer cells. The results suggest that miR‐142‐3p could be a good candidate for the targeted therapy of breast cancer, especially for the invasive type. Abstract Background Breast cancer is the most common type of cancer among women, and despite improved treatments, it remains a major challenge. However, improved mechanistic insight may lead to novel therapeutic strategies. miR‐142‐3p belongs to the miR‐142 family and is involved in pathogenesis and metastasis of various types of malignancies by targeting several important messenger RNAs (mRNAs) including Bach‐1. This is especially true for breast cancer, where Bach‐1 is involved in the metastatic spread by deregulation of metastasis‐associated genes. Methods In this study, we collected 24 breast cancer tissues with 24 adjusted normal tissues to measure the expression levels of miR‐142‐3p and Bach‐1 mRNA using quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR) and IHC. miR‐142‐3p targeting of Bach‐1 expression in MCF‐7 and MDA‐MB‐468 breast cancer cells was evaluated using bioinformatics, qRT‐PCR and western blot analyses. The cellular proliferation, invasion, and migration were assessed by MTT, transwell matrigel and wound healing assay and the EMT‐associated proteins C‐X‐C chemokine receptor type 4 (CXCR‐4), matrix metalloproteinase‐9 (MMP9), and vascular endothelial growth factor receptor (VEGFR) were analyzed by western blot analysis. Also, the expression levels of tumor suppressors including miR‐330, miR‐145, and miR‐34a were estimated by qRT‐PCR. Results Analysis of paired specimens of primary malignant and normal tissues showed that miR‐142‐3p was downregulated, while Bach‐1 mRNA and protein both were overexpressed in the breast cancer tumors. This inverse relationship was confirmed by cell line experiments demonstrating that miR‐142‐3p expression reduced Bach‐1 mRNA levels. Furthermore, replacement of miR‐142‐3p could inhibit the proliferation, invasion, and migration in breast cancer potentially by targeting of Bach‐1 mRNA and subsequent inhibition of CXCR4, MMP9, and VEGFR protein expressions. In addition, induction of miR‐142‐3p could upregulate tumor suppressor miRNAs, including miR‐330, miR‐145, and miR34a. Conclusion For the first time, our results revealed that miR‐142‐3p could target Bach‐1in breast cancer cells leading to the reduction of EMT‐related proteins and reduced cell proliferation, invasion, and migration. The results also demonstrated that miR‐142‐3p could regulate important tumor suppressor miRNAs in breast cancer cells. In conclusion, our results suggest that miR‐142‐3p could be a good candidate for the targeted therapy of breast cancer, especially for the invasive type. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27670   open full text
• A novel multidimensional signature predicts prognosis in hepatocellular carcinoma patients.
  Song Wang, Jian‐Hua Zhang, Huan Wang, Lu Yang, Shuai Hong, Bo Yu, Jin‐Cheng Guo, Jing Liu, Yan‐Bing Zhu.
  Journal of Cellular Physiology. November 27, 2018
  --- - "\nUsing bioinformatics analysis methods including Cox’s proportional hazards regression analysis, the random survival forest algorithm, Kaplan–Meier and receiver operating characteristic curve analysis, we mined the gene expression profiles of 469 hepatocellular carcinoma (HCC) patients from The Cancer Genome Atlas (n = 379) and Gene Expression Omnibus (GSE14520; \nn = 90) public database and found a prognostic protein‐coding gene‐microRNA signature that predicts survival in patients with HCC patients. The signature has a bright clinical significance to be a potential prognostic biomarker.\n\n\n\n\n\nAbstract\nThe abnormal expression of microRNAs (miRNAs) or protein‐coding genes (PCGs) have been found to be associated with the prognosis of hepatocellular carcinoma (HCC) patients. Using bioinformatics analysis methods including Cox’s proportional hazards regression analysis, the random survival forest algorithm, Kaplan–Meier, and receiver operating characteristic (ROC) curve analysis, we mined the gene expression profiles of 469 HCC patients from The Cancer Genome Atlas (n = 379) and Gene Expression Omnibus (GSE14520; \nn = 90) public database. We selected a signature comprising one protein‐coding gene (PCG; DNA polymerase μ) and three miRNAs (hsa‐miR‐149‐5p, hsa‐miR‐424‐5p, hsa‐miR‐579‐5p) with highest accurate prediction (area under the ROC curve [AUC] = 0.72; \nn = 189) from the training data set. The signature stratified patients into high‐ and low‐risk groups with significantly different survival (median 27.9 vs. 55.2 months, log‐rank test, \np < 0.001) in the training data set, and its risk stratification ability were validated in the test data set (median 47.4 vs. 84.4 months, log‐rank test, \np = 0.03) and an independent data set (median 31.0 vs. 46.0 months, log‐rank test, \np = 0.01). Multivariable Cox regression analysis showed that the signature was an independent prognostic factor. And the signature was proved to have a better survival prediction power than tumor–node–metastasis (TNM) stage (AUC\nsignature = 0.72/0.64/0.62 vs. AUC\nTNM = 0.65/0.61/0.61; \np < 0.05). Moreover, we validated the expression of these prognostic genes from the PCG‐miRNA signature in Huh‐7 cell by real‐time polymerase chain reaction. In conclusion, we found a signature that can predict survival of HCC patients and serve as a prognostic marker for HCC." - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27818   open full text
• Human breast cancer decellularized scaffolds promote epithelial‐to‐mesenchymal transitions and stemness of breast cancer cells in vitro.
  Gang Liu, Biao Wang, Shubin Li, Qin Jin, Yanfeng Dai.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- In our study, human breast cancer biopsies were decellularized by the detergent technique with sodium lauryl ether sulfate (SLES) solution for the first time. Scaffolds seeded with MCF‐7 cells demonstrated the process of cell recellularization in vitro, with increased cell migration, proliferation, and epithelial‐to‐mesenchymal transition (EMT) process. When treated with 5‐fluorouracil (5‐Fu), the expressions of stem cell markers, including Oct4, Sox2, and CD49F, were maximally maintained in the recellularized scaffold with decreased apoptosis rates compared with monolayer cells. Abstract Breast cancer, with unsatisfactory survival rates, is the leading cause of cancer‐related death in women worldwide. Recent advances in the genetic basis of breast cancer have benefitted the development of gene‐based medicines and therapies. Tissue engineering technologies, including tissue decellularizations and reconstructions, are potential therapeutic alternatives for cancer research and tissue regeneration. In our study, human breast cancer biopsies were decellularized by a detergent technique, with sodium lauryl ether sulfate (SLES) solution, for the first time. And the decellularization process was optimized to maximally maintain tissue microarchitectures and extracellular matrix (ECM) components with minimal DNA compounds preserved. Histology analysis and DNA quantification results confirmed the decellularization effect with maximal genetic compounds removal. Quantification, immunofluorescence, and histology analyses demonstrated better preservation of ECM components in 0.5% SLES‐treated scaffolds. Scaffolds seeded with MCF‐7 cells demonstrated the process of cell recellularization in vitro, with increased cell migration, proliferation, and epithelial‐to‐mesenchymal transition (EMT) process. When treated with 5‐fluorouracil, the expressions of stem cell markers, including Oct4, Sox2, and CD49F, were maximally maintained in the recellularized scaffold with decreased apoptosis rates compared with monolayer cells. These results showed that the decellularized breast scaffold model with SLES treatments would help to simulate the pathogenesis of breast cancer in vitro. And we hope that this model could further accelerate the development of effective therapies for breast cancer and benefit drug screenings. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27630   open full text
• Identification of hsa‐mir‐34a, hsa‐mir‐124, and hsa‐mir‐204 as signatures for cataract.
  Ying‐Li Wei, Hao Sun.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Three key microRNAs (miRNAs) related to cataract pathogenesis are identified, and the corresponding target genes and transcription factors are also analyzed. Moreover, the potential functions of the target genes are revealed. The results in the study may contribute to the targeted therapy of cataract through miRNAs. Abstract Cataract, one of the common age‐related diseases that remains the leading cause of blindness worldwide. There have been studies pertaining to microRNA (miRNA) effects on cataract pathogenesis. However, the results were different from each other due to the influence of sequencing platform, sample selection, filter conditions, and so on. This study aims to explore the key miRNAs underlying the pathogenesis of cataract and find prevention and treatment methods. First, six representative cataract miRNA datasets are selected and the key miRNAs are determined. The target genes of these key miRNAs are predicted using the TargetScan software, and then, the functional enrichment and transcription factor binding analysis of these target genes are completed, respectively. Three signature miRNAs associated with cataract are finally identified. Among them, hsa‐mir‐34a seems to be significantly upregulated, while hsa‐mir‐124 and hsa‐mir‐204 are significantly downregulated. The target genes of the signature miRNAs are mainly enriched in hsa04015:Rap1 signaling pathway, hsa04142:lysosome, and hsa04360:axon guidance, indicating that these pathways may be the crucial metabolic pathways. To summarize, three key miRNAs related to cataract pathogenesis are identified, and the corresponding target genes and transcription factors are also analyzed. Moreover, the potential functions of the target genes are revealed. The results of this study may contribute to the targeted therapy of cataract through miRNAs. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27741   open full text
• Comparative study of laparoscopic‐assisted and open total gastrectomy for Siewert Types II and III adenocarcinoma of the esophagogastric junction.
  Jianchu Wang, Jin‐Cheng Wang, Bin Song, Xu‐Dong Dai, Xiao‐Yu Zhang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 Abstract Background The potential advantages of laparoscopic‐assisted total gastrectomy (LATG) compared with open total gastrectomy (OTG) for Siewert Types II and III adenocarcinoma of the esophagogastric junction (AEJ) are not very clear. Thus, the aim of this study was to investigate the surgical outcomes and potential advantages of LATG for Siewert Types II and III AEJ. Methods The clinical data of 75 patients (32 for LATG and 43 for OTG) with Siewert II or III AEJ from August 2009 to February 2014 were analyzed retrospectively. Patients were followed up by telephone or out‐patient examination till August 2015. Results Two groups of patients were successfully performed with no perioperative death. The mean operation time was 3.23 ± 0.35 hr in LATG group, longer than the OTG group 2.83 ± 0.51 hr. The mean intraoperative bleeding was 122.7 ± 50.6 ml, less than the OTG group 219.2 ± 85.2 ml. The analgesics use was 3.00 ± 0.67 times in the LATG group, less than the OTG group 3.43 ± 1.03 times. The gastrointestinal function recovery time was 2.69 ± 0.46 days in the LATG group, shorter than the OTG group 3.42 ± 0.86 days. The mean postoperative hospital stay was 12.94 + 2.76 days in the LATG group, less than the OTG group 14.57 + 2.35 days (p < 0.05). Conclusions LATG and OTG had no significant difference for Siewert II and III AEJ in terms of radical resection and tumor recurrence, but LATG is worthy to be promoted with less bleeding, less postoperative pain, faster recovery of gastrointestinal function, and shorter hospital stay. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27777   open full text
• SP1, MYC, CTNNB1, CREB1, JUN genes as potential therapy targets for neuropathic pain of brain.
  Xue‐Tao Yan, Yong Xu, Xiao‐Li Cheng, Xiang‐Hu He, Yu Wang, Wen‐Zhong Zheng, Ying Zhao, Hu Chen, Yan‐Lin Wang.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Our results suggested a potential application of SP1, MYC, CTNNB1, CREB1, and JUN genes as prognostic biomarkers in the clinical management of neuropathic pain, and two most important signaling pathways, immune response and reception reactions, were found to be closely related to the occurrence and development of neuropathic pain. The more specific mechanism of these genes with the brain tissue related to neuropathic pain stimulation needs further research. Abstract Neuropathic pain (NP) may cause serious brain diseases, but the genes associated with the metabolic pathway and transcript factors of NP remain unclear. This study is aimed to identify the therapy target genes for NP and to investigate the metabolic pathways and transcript factors associated with NP. The differentially expressed genes of three brain tissues (nucleus accumbens, periaqueductal gray, and prefrontal cortex) dealt with NP stimulation were analyzed. Besides, The Database for Annotation, Visualization, and Integrated Discovery and Tfacts datasets were used in the analysis of the genes related to the metabolic pathway and transcript factors of the brain. Eight genes were found to coexpress in all three tissues. A functional enrichment analysis showed that the upregulated genes were mostly enriched in pathways as inflammatory response, calcium‐mediated signaling, cytokine‐cytokine receptor interaction, and extracellular matrix (ECM)‐receptor interaction, whereas the downregulated genes were mostly enriched in pathways as phospholipid metabolic processes, positive regulation of protein kinase B signaling, and metabolism of xenobiotics by cytochrome P450. Finally, 135 and 98 transcript factors genes were upregulated and downregulated, among which SP1, MYC, CTNNB1, CREB1, JUN were identified as the most critical genes because the number of up‐ and downregulated gene ranked at the top. In conclusion, the pathways of immune response and cytokine‐cytokine receptor interaction were determined as the main metabolic pathways of NP affecting the brain, and SP1, MYC, CTNNB1, CREB1, JUN genes were recognized as the most enriched genes in this process, which may provide evidence for the diagnosis and treatment research of neuropathic pain. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27413   open full text
• Synergistic anticancer action of quercetin and curcumin against triple‐negative breast cancer cell lines.
  Sai Kundur, Amrita Prayag, Priyanga Selvakumar, Hung Nguyen, Lloyd McKee, Clairissa Cruz, Asha Srinivasan, Sunday Shoyele, Ashakumary Lakshmikuttyamma.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Women with the breast cancer type 1 susceptibility protein (BRCA1) mutation and loss of BRCA1 expression are reported to have an increased risk of triple‐negative breast cancer (TNBC). We detected that quercetin and curcumin dose‐dependently enhanced BRCA1 expression. Quercetin and curcumin appeared to induce BRCA1 promoter histone acetylation. Our present study concluded that the combination treatment of quercetin and curcumin acts synergistically to induce anticancer activity against TNBC cells by modulating tumor suppressor genes. Abstract Women with the breast cancer type 1 susceptibility protein (BRCA1) mutation and loss of BRCA1 expression are reported to have an increased risk of triple‐negative breast cancer (TNBC). Targeting BRCA1 modulation might offer a therapeutic option to treat TNBC patients. Our studies detected that BRCA1 is poorly expressed in TNBC cell lines and highly expressed in ER+ breast cancer cell lines. To modulate BRCA1 expression, we tested two different dietary components to find out if any would induce tumor suppressor genes. We detected that quercetin and curcumin dose‐dependently enhanced the BRCA1 expression. Further, a synergistic action of quercetin and curcumin was observed in modulating the BRCA1 level and in inhibiting the cell survival and migration of TNBC cell lines. Quercetin and curcumin appeared to induce BRCA1 promoter histone acetylation. Furthermore, BRCA1 knockdown induced cell survival and cell migration in ER + cells were significantly decreased by the combined treatment of quercetin and curcumin. Our present study concluded that the combination treatment of quercetin and curcumin acts synergistically to induce anticancer activity against TNBC cells by modulating tumor suppressor genes. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27761   open full text
• Induced pluripotent stem cell‐derived extracellular vesicles: A novel approach for cell‐free regenerative medicine.
  Behnaz Taheri, Masoud Soleimani, Sedigheh Fekri Aval, Elahe Esmaeili, Zahra Bazi, Nosratollah Zarghami.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- Extracellular vesicles contribute to the therapeutic effect of the stem cells, including stimulation of organ repair. The discovery of induced pluripotent stem cells represented a promising source for autologous extracellular vesicles generation to use in regenerative medicine. Abstract In recent years, induced pluripotent stem cells (iPSCs) have been considered as a promising approach in the field of regenerative medicine. iPSCs can be generated from patients’ somatic cells and possess the potential to differentiate, under proper conditions, into any cell type. However, the clinical application of iPS cells is restricted because of their tumorigenic potential. Recent studies have indicated that stem cells exert their therapeutic benefit via a paracrine mechanism, and extracellular vesicles have been demonstrated that play a critical role in this paracrine mechanism. Due to lower immunogenicity, easier management, and presenting no risk of tumor formation, in recent years, researchers turned attention to exosomes as potential alternatives to whole‐cell therapy. Application of exosomes derived from iPSCs and their derived precursor provides a promising approach for personalized regenerative medicine. This study reviews the physiological functions of extracellular vesicles and discusses their potential therapeutic benefit in regenerative medicine. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27775   open full text
• Glucagon attenuates lipid accumulation in cow hepatocytes through AMPK signaling pathway activation.
  Yu Li, Hongyan Ding, Jihong Dong, Sajid Ur Rahman, Shibin Feng, Xichun Wang, Jinjie Wu, Zhe Wang, Guowen Liu, Xiaobing Li, Xinwei Li.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- The ketotic cows displayed hepatic lipid metabolic disorder and high blood concentration of glucagon. The present study indicates that glucagon activates the AMPK signalling pathway to increase lipid oxidation and VLDL assembly and decrease lipid synthesis in cow hepatocytes, thereby reducing liver fat accumulation. Abstract The ketotic cows displayed hepatic lipid metabolic disorder and high blood concentration of glucagon. Importantly, adenosine monophosphate‐activated protein kinase (AMPK) signaling pathway plays an important role in the hepatic lipid homeostasis. Therefore, the aim of this study was to investigate the effect of glucagon on AMPK pathway and its underlying mechanism on lipid metabolism in cow hepatocytes. Cow hepatocytes were cultured and treated with glucagon and AMPK inhibitor (BML‐275). The results showed that glucagon significantly promoted the expression of glucagon receptor and increased the phosphorylation level and activity of AMPKα. Activated AMPKα increased the expression level and transcriptional activity of peroxisome proliferator‐activated receptor α, which further increased the expression of fatty acid oxidation genes and lipid oxidation. Furthermore, activated AMPKα inhibited the expression level and transcriptional activity of sterol regulatory element binding protein‐1c and carbohydrate response element binding protein, which decreased the expression of lipogenic genes, thereby decreasing lipid synthesis. In addition, glucagon also increased the expression of very‐low‐density lipoprotein (VLDL) assembly to export intracellular triglycerides (TG). Consequently, the content of intracellular TG was significantly decreased in cow hepatocytes. These results indicate that glucagon activates the AMPK signaling pathway to increase lipid oxidation and VLDL assembly and decrease lipid synthesis in cow hepatocytes, thereby reducing liver fat accumulation. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27258   open full text
• Comparative transcriptome analysis reveals PERP upregulated during Salmonella Enteritidis challenge in laying ducks.
  Yu Zhang, Tian‐tian Gu, Yang Chen, Yu Huang, Jinping Du, Lizhi Lu, Guo‐qiang Zhu, Qi Xu, Guo‐hong Chen.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2- The PERP overexpression resulted in an increased induction of p53, MDM2, caspase‐3, and Bcl‐2 during SE infection in dGCs, whereas an opposite trend was observed with PERP knockdown. Duck PERP, as a negative regulator, was involved in the ovarian local immune niche through a p53 signaling pathway that promoted cell apoptosis, as well as SE adhesion and invasion during SE infection. Abstract Salmonella Enteritidis (SE) can be transmitted to eggs through cecum or the ovary from infected layers and causes food poisoning in humans. The mechanism of cecal transmission has been extensively studied. However, the mechanism and route of transovarian transmission of SE remain unclear. In this study, the ducks were orally inoculated with SE, and the ovarian follicles and stroma were collected to detect SE infection. The immune responses were triggered and the innate and adaptive immune genes (TLR4, NOD1, AvβD7, and IL‐1β) were upregulated significantly during the SE challenge. Moreover, the ovary tissues (small follicle and stroma) of susceptible and resistant–laying ducks were performed by RNA sequencing. We obtained and identified 23 differentially expressed genes (DEGs) between susceptible and resistant–laying ducks in both small follicle and stroma tissues ( p < 0.05). The DEGs were predominately identified in the p53 signaling pathway. The expression of key genes (p53, MDM2, PERP, caspase‐3, and Bcl‐2) involved in the signaling pathway was significantly higher in granulosa cells (dGCs) from SE‐infected ducks than those from uninfected ducks. Moreover, the overexpression of PERP resulted in further induction of p53, MDM2, caspase‐3, and Bcl‐2 during SE infection in dGCs. Whereas, an opposite trend was observed with the knockdown of PERP. Besides, it is further revealed that the PERP could enhance cell apoptosis, SE adhesion, and SE invasion in SE‐infected dGCs overexpression. Altogether, our results demonstrate the duck PERP involved in the ovarian local immune niche through p53 signaling pathway in dGCs challenged with SE. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27790   open full text
• The role of nitric oxide signaling in renoprotective effects of hydrogen sulfide against chronic kidney disease in rats: Involvement of oxidative stress, autophagy and apoptosis.
  Mohammad Khabbaz Shirazi, Asaad Azarnezhad, Mohammad Foad Abazari, Mansour Poorebrahim, Pegah Ghoraeian, Nima Sanadgol, Hanieh Bokharaie, Sahar Heydari, Amin Abbasi, Sahra Kabiri, Maryam Nouri Aleagha, Seyed Ehsan Enderami, Amir Savar Dashtaki, Hassan Askari.
  Journal of Cellular Physiology. November 27, 2018
  --- - |2 Abstract The interplay between H2S and nitric oxide (NO) is thought to contribute to renal functions. The current study was designed to assess the role of NO in mediating the renoprotective effects of hydrogen sulfide in the 5/6 nephrectomy (5/6 Nx) animal model. Forty rats were randomly assigned to 5 experimental groups: (a) Sham; (b) 5/6 Nx; (c) 5/6Nx+sodium hydrosulfide‐a donor of H 2S, (5/6Nx+sodium hydrosulfide [NaHS]); (d) 5/6Nx+NaHS+ L‐NAME (a nonspecific nitric oxide synthase [NOS] inhibitor); (e) 5/6Nx+NaHS+aminoguanidine (a selective inhibitor of inducible NOS [iNOS]). Twelve weeks after 5/6 Nx, we assessed the expressions of iNOS and endothelial NOS (eNOS), oxidative/antioxidant status, renal fibrosis, urine N‐acetyl‐b‐glucosaminidase (NAG) activity as the markers of kidney injury and various markers of apoptosis, inflammation, remodeling, and autophagy. NaHS treatment protected the animals against chronic kidney injury as depicted by improved oxidative/antioxidant status, reduced apoptosis, and autophagy and attenuated messenger RNA (mRNA) expression of genes associated with inflammation, remodeling, and NAG activity. Eight weeks Nω‐nitro‐l‐arginine methyl ester ( L‐NAME) administration reduced the protective effects of hydrogen sulfide. In contrast, aminoguanidine augmented the beneficial effects of hydrogen sulfide. Our finding revealed some fascinating interactions between NO and H 2S in the kidney. Moreover, the study suggests that NO, in an isoform‐dependent manner, can exert renoprotective effects in 5/6 Nx model of CKD. - 'Journal of Cellular Physiology, EarlyView. '

  November 27, 2018   doi: 10.1002/jcp.27797   open full text
• Global estimate of gastric cancer in Helicobacter pylori–infected population: A systematic review and meta‐analysis.
  Ali Pormohammad, Naser Mohtavinejad, Pourya Gholizadeh, Hossein Dabiri, Alireza Salimi Chirani, Ali Hashemi, Mohammad Javad Nasiri.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- The frequency of gastric cancer (GC) among H. pylori–infected population varied markedly across countries. The highest rate of GC was observed in H. pylori–infected individuals from Asian countries. The frequency of GC was relatively high in H. pylori–infected population in the world. However, the eradication of H. pylori might be a promising strategy for GC prevention, especially in high‐risk populations such as Asian countries. There is information regarding the rates of gastric cancer (GC) in different populations and the important role of Helicobacter pylori in GC development; however, no comprehensive study has yet been performed to investigate the prevalence of GC in H. pylori–infected patients. PubMed, Embase, and Cochrane Library through January 1, 2000 were searched without language restrictions. Quality of included studies was assessed with a critical appraisal checklist recommended by the Joanna Briggs Institute. All of the analyses were conducted using Comprehensive Meta‐Analysis Software Version 2.0 and Stata 14.0. Forty‐four studies from 17 countries were included. The pooled frequency of GC was 17.4% (95% confidence interval: 16.4–18.5) in H. pylori–infected population. The frequency of GC among H. pylori–infected population varied markedly across countries. The highest rate of GC was observed in H. pylori–infected individuals from Asian countries. The frequency of GC was relatively high in H. pylori–infected population in the world. However, the eradication of H. pylori might be a promising strategy for GC prevention, especially in high‐risk populations such as Asian countries. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1208-1218, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27114   open full text
• Role and translational implication of galectins in arthritis pathophysiology and treatment: A systematic literature review.
  Francesca Salamanna, Francesca Veronesi, Antonio Frizziero, Milena Fini.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Galectins are members of the animal lectin family that bind to the β‐galactoside‐containing carbohydrate moieties of glycoconjugates. This descriptive systematic literature review summarizes in vitro, in vivo, and clinical studies that analyzed and examined the role and mechanism of action of galectins in arthritis to highlight and clarify their possible translation implication. Manipulation of certain galectins can suppress or aggravate disease symptoms in arthritis animal models, demonstrating the therapeutic potential of galectins for the treatment of rheumatoid arthritis and osteoarthritis. Galectins are members of the animal lectin family that bind to the β‐galactoside‐containing carbohydrate moieties of glycoconjugates. They seem to have an important role in the pathophysiology of several diseases, including arthritis. Osteoarthritis (OA) and rheumatoid arthritis (RA) are chronic conditions with few or no available therapies. In this context, galectins could provide a novel opportunity, but the precise role and mechanism of their involvement in arthritis are still not fully understood. This descriptive systematic literature review summarizes in vitro, in vivo, and clinical studies that analyzed and examined the role and mechanism of action of galectins in arthritis to highlight and clarify their possible translation implication. This review yielded promising evidence that individual galectins, in particular galectin‐1, ‐3, and ‐9, could play positive or negative roles in the pathogenesis of arthritis, especially in RA and OA. It also emphasized the cell‐dependent role of these galectins. This is particularly true for galectin‐1, which was shown to have a protective anti‐inflammatory role in RA, while it seemed to be associated with cartilage degeneration in OA. In summary, this review underlined that manipulation of certain galectins can suppress or aggravate disease symptoms in arthritis animal models, demonstrating the therapeutic potential of galectins for the treatment of RA and OA. Nevertheless, despite the fact that galectin therapy and therapies acting on galectin expression seem to be an interesting and important opportunity for research, we highlighted that further investigation is necessary to carefully evaluate their potential clinical implications in arthritis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1588-1605, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27026   open full text
• A cellular perspective of adipogenesis transcriptional regulation.
  Walid Kuri‐Harcuch, Cristina Velez‐delValle, Alfredo Vazquez‐Sandoval, Claudia Hernández‐Mosqueira, Veronica Fernandez‐Sanchez.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this review, we examine the research bearing on the control of adipose cell differentiation, some of it dating back to the early days when Howard Green and his group described the preadipocyte cell lines 3T3‐L1 and 3T3 F442A and also concentrated our attention on research published during the last few years, emphasizing data described to regulate adipose differentiation, outside of those that are part of the canonical adipogenic transcriptional cascade. Adipose cells store lipids in the cytoplasm and signal systemically through secretion of adipokines and other molecules that regulate body energy metabolism. Differentiation of fat cells and its regulation has been the focus of extensive research since the early 1970s. In this review, we had attempted to examine the research bearing on the control of adipose cell differentiation, some of it dating back to the early days when Howard Green and his group described the preadipocyte cell lines 3T3‐L1 and 3T3‐F442A during 1974–1975. We also concentrated our attention on research published during the last few years, emphasizing data described on transcription factors that regulate adipose differentiation, outside of those that were reported earlier as part of the canonical adipogenic transcriptional cascade, which has been the subject of ample reviews by several groups of researchers. We focused on the studies carried out with the two preadipocyte cell culture models, the 3T3‐L1 and 3T3‐F442A cells that have provided essential data on adipose biology. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1111-1129, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27060   open full text
• A review of the molecular mechanisms of hyperglycemia‐induced free radical generation leading to oxidative stress.
  Habib Yaribeygi, Stephen L. Atkin, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Studies have shown that many of the diabetes complications result from excessive free radical generation and oxidative stress, and it has been shown that chronic hyperglycemia is a potent inducer for free radical production, generated through several pathways and triggering multiple molecular mechanisms. An understanding of these processes may help us to improving our preventive or therapeutic strategies. In this review, the major molecular pathways involved in free radical generation induced by hyperglycemia are described. Abstract The prevalence of diabetes is growing worldwide with an increasing morbidity and mortality associated with the development of diabetes complications. Free radical production is a normal biological process that is strictly controlled and has been shown to be important in normal cellular homeostasis, and in the bodies response to pathogens. However, there are several mechanisms leading to excessive free radical production that overcome the normal protective quenching mechanisms. Studies have shown that many of the diabetes complications result from excessive free radical generation and oxidative stress, and it has been shown that chronic hyperglycemia is a potent inducer for free radical production, generated through several pathways and triggering multiple molecular mechanisms. An understanding of these processes may help us to improving our preventive or therapeutic strategies. In this review, the major molecular pathways involved in free radical generation induced by hyperglycemia are described. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1300-1312, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27164   open full text
• The potential prognostic and therapeutic application of tissue and circulating microRNAs in cervical cancer.
  Malihe Hasanzadeh, Mehraneh Movahedi, Marzieh Rejali, Faezeh Maleki, Mehrdad Moetamani‐Ahmadi, Sima Seifi, Zeinab Hosseini, Majid Khazaei, Forouzan Amerizadeh, Gordon A. Ferns, Majid Rezayi, Amir Avan.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Exosomes and microRNAs. Exosomes contain genetic information of their cellular source, and circulate throughout body, indictaing its values as noninvasive biomarkers. Abstract Cervical cancer (CC) is a common malignancy in women and a major cause of cancer‐related mortality globally. Some novel biomarkers may enable the early diagnosis and monitoring of CC. MicroRNAs (miRNAs) are small noncoding RNAs that control gene translation at a posttranscriptional level. Hence the deregulation of these molecules can cause many diseases. There appears to be an association between aberrant miRNA expression and CC, but the molecular mechanisms involved in the development of CC remain unknown. The upregulation of some circulating miRNAs, for example, miRNA‐20a, miRNA‐203, miRNA‐21, miRNA‐205, miRNA‐218, and miR‐485‐5, as well as tissue‐specific miRNAs, for example, miR‐7, miR‐10a, miR‐17‐5p, miR‐135b, miR‐149, and miR‐203 have been found in patients with CC. There is also growing evidence for the importance of miRNAs in the development of drug resistance. This review therefore highlights recently published preclinical and clinical investigation performed on tissue specific and circulating miRNAs, as potential biomarkers for the detection of patients at early stages of CC, in the prediction of prognosis, and monitoring of their response to therapy. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1289-1294, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27160   open full text
• Anticancer properties of tocotrienols: A review of cellular mechanisms and molecular targets.
  Marina Montagnani Marelli, Monica Marzagalli, Fabrizio Fontana, Michela Raimondi, Roberta Manuela Moretti, Patrizia Limonta.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Vitamin E TTs have gained interest due to their health benefits in chronic diseases. In particular, TTs were reported to exert significant antitumor activity in a wide range of cancer cells, by counteracting cell growth or proliferation, metastasis and angiogenesis. Recent studies pointed out that TTs exert a synergistic antitumor effect on cancer cells when given in combination with either standard antitumor agents or natural compounds with anticancer activity. These results, together with the safety of TT administration in healthy subjects, suggest that these compounds might represent a new chemopreventive or anticancer treatment strategy. Clinical trials aimed at confirming this antitumor activity of TTs are needed. Vitamin E is composed of two groups of compounds: α‐, β‐, γ‐, and δ‐tocopherols (TPs), and the corresponding unsaturated tocotrienols (TTs). TTs are found in natural sources such as red palm oil, annatto seeds, and rice bran. In the last decades, TTs (specifically, γ‐TT and δ‐TT) have gained interest due to their health benefits in chronic diseases, based on their antioxidant, neuroprotective, cholesterol‐lowering, anti‐inflammatory activities. Several in vitro and in vivo studies pointed out that TTs also exert a significant antitumor activity in a wide range of cancer cells. Specifically, TTs were shown to exert antiproliferative/proapoptotic effects and to reduce the metastatic or angiogenic properties of different cancer cells; moreover, these compounds were reported to specifically target the subpopulation of cancer stem cells, known to be deeply involved in the development of resistance to standard therapies. Interestingly, recent studies pointed out that TTs exert a synergistic antitumor effect on cancer cells when given in combination with either standard antitumor agents (i.e., chemotherapeutics, statins, “targeted” therapies) or natural compounds with anticancer activity (i.e., sesamin, epigallocatechin gallate (EGCG), resveratrol, ferulic acid). Based on these observations, different TT synthetic derivatives and formulations were recently developed and demonstrated to improve TT water solubility and to reduce TT metabolism in cancer cells, thus increasing their biological activity. These promising results, together with the safety of TT administration in healthy subjects, suggest that these compounds might represent a new chemopreventive or anticancer treatment (i.e., in combination with standard therapies) strategy. Clinical trials aimed at confirming this antitumor activity of TTs are needed. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1147-1164, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27075   open full text
• Cell communication by tunneling nanotubes: Implications in disease and therapeutic applications.
  Rahul Mittal, Elisa Karhu, Jay‐Shing Wang, Stefanie Delgado, Ryan Zukerman, Jeenu Mittal, Vasanti M. Jhaveri.
  Journal of Cellular Physiology. November 20, 2018
  --- - | A schematic representation of various cellular cargoes transported by tunneling nanotubes (TNTs). nanotubes Abstract Intercellular communication is essential for the development and maintenance of multicellular organisms. Tunneling nanotubes (TNTs) are a recently recognized means of long and short distance communication between a wide variety of cell types. TNTs are transient filamentous membrane protrusions that connect cytoplasm of neighboring or distant cells. Cytoskeleton fiber‐mediated transport of various cargoes occurs through these tubules. These cargoes range from small ions to whole organelles. TNTs have been shown to contribute not only to embryonic development and maintenance of homeostasis, but also to the spread of infectious particles and resistance to therapies. These functions in the development and progression of cancer and infectious disease have sparked increasing scrutiny of TNTs, as their contribution to disease progression lends them a promising therapeutic target. Herein, we summarize the current knowledge of TNT structure and formation as well as the role of TNTs in pathology, focusing on viral, prion, and malignant disease. We then discuss the therapeutic possibilities of TNTs in light of their varied functions. Despite recent progress in the growing field of TNT research, more studies are needed to precisely understand the role of TNTs in pathological conditions and to develop novel therapeutic strategies. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1130-1146, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27072   open full text
• Liposomal nanocarriers for statins: A pharmacokinetic and pharmacodynamics appraisal.
  Anis Askarizadeh, Alexandra E. Butler, Ali Badiee, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 Abstract Statins, inhibitors of 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase, are a well‐known class of drug with beneficial therapeutic effects in cardiovascular disease and lipid disorders and have potential use against cancer. However, the bioavailability of statins is hampered due to low aqueous solubility and rapid metabolism. To improve pharmacokinetic profiles of statins, development of drug delivery systems is promising. Hence, the use of liposomes for selective delivery of statins to a selected site or for bioavailability enhancement is an effective strategy to increase statin therapeutic effects. Moreover, liposomal delivery can reduce the required dose of statins especially in terms of antitumor effects. Liposomes, because of their unique properties and biphasic and amphiphilic nature, have attracted much interest and can be considered as a suitable choice for delivery of both hydrophilic and lipophilic statins. In this review article, we focus on liposomes and evaluate the effects of different liposomal delivery systems, based on differences in size, phospholipid composition, circulation half‐life, and cholesterol content, on statin function. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1219-1229, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27121   open full text
• Regulation of tumor angiogenesis by microRNAs: State of the art.
  Nasser H. Goradel, Nejad Mohammadi, Hamed Haghi‐Aminjan, Bagher Farhood, Babak Negahdari, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- This review focuses on tumor angiogenesis regulation by microRNAs and the mechanism underlying this regulation. MicroRNAs (miRNAs, miRs) are small (21–25 nucleotides) endogenous and noncoding RNAs involved in many cellular processes such as apoptosis, development, proliferation, and differentiation via binding to the 3′‐untranslated region of the target mRNA and inhibiting its translation. Angiogenesis is a hallmark of cancer, which provides oxygen and nutrition for tumor growth while removing deposits and wastes from the tumor microenvironment. There are many angiogenesis stimulators, among which vascular endothelial growth factor (VEGF) is the most well known. VEGF has three tyrosine kinase receptors, which, following VEGF binding, initiate proliferation, invasion, migration, and angiogenesis of endothelial cells in the tumor environment. One of the tumor microenvironment conditions that induce angiogenesis through increasing VEGF and its receptors expression is hypoxia. Several miRNAs have been identified that affect different targets in the tumor angiogenesis pathway. Most of these miRNAs affect VEGF and its tyrosine kinase receptors expression downstream of the hypoxia‐inducible Factor 1 (HIF‐1). This review focuses on tumor angiogenesis regulation by miRNAs and the mechanism underlying this regulation. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1099-1110, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27051   open full text
• PD‐1/PD‐L1 immune checkpoint: Potential target for cancer therapy.
  Fatemeh K. Dermani, Pouria Samadi, Golebagh Rahmani, Alisa K. Kohlan, Rezvan Najafi.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this review, we highlight a brief overview of the molecular and biochemical events that are regulated by the PD‐1 and PD‐L1 interaction in various cancers. Abstract Recent studies show that cancer cells are sometimes able to evade the host immunity in the tumor microenvironment. Cancer cells can express high levels of immune inhibitory signaling proteins. One of the most critical checkpoint pathways in this system is a tumor‐induced immune suppression (immune checkpoint) mediated by the programmed cell death protein 1 (PD‐1) and its ligand, programmed death ligand 1 (PD‐L1). PD‐1 is highly expressed by activated T cells, B cells, dendritic cells, and natural killer cells, whereas PD‐L1 is expressed on several types of tumor cells. Many studies have shown that blocking the interaction between PD‐1 and PD‐L1 enhances the T‐cell response and mediates antitumor activity. In this review, we highlight a brief overview of the molecular and biochemical events that are regulated by the PD‐1 and PD‐L1 interaction in various cancers. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1313-1325, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27172   open full text
• Mesenchymal stem cells as the game‐changing tools in the treatment of various organs disorders: Mirage or reality?
  Faroogh Marofi, Ghasem Vahedi, Ali hasanzadeh, Sadegh Salarinasab, Pishva Arzhanga, Bahareh Khademi, Majid Farshdousti Hagh.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Recently a growing attention in scientific community has been gathered on potential application of mesenchymal stem cells (MSCs) in various fields of medicine. Various useful properties of MSCs including tropism toward tumor/injury site(s), weakly immunogenic, production of anti‐inflammatory molecules, and safety against normal tissues have made them prone for regenerative medicine, targeted therapy and treating injured tissues, and immunological abnormalities. Abstract Recently a growing attention in scientific community has been gathered on potential application of mesenchymal stem cells (MSCs) in various fields of medicine. Owing to the fact that they can be easily isolated from different sources, and simply proliferated in large quantities while keeping their original biological characteristics, they can be successfully used as cell‐based therapeutics. Engineering MSCs and other type of stem cells to be carriers of therapeutic agents is a new tactic in the targeted gene and cell therapy of cancers and degenerative diseases. Various useful properties of MSCs including tropism toward tumor/injury site(s), weakly immunogenic, production of anti‐inflammatory molecules, and safety against normal tissues have made them prone for regenerative medicine, targeted therapy and treating injured tissues, and immunological abnormalities. In this review, we introduce latest advances, methods, and applications of MSCs in gene therapy of various malignant organ disorders. Additionally, we will cover the problems and challenges which researchers have faced with when trying to translate their basic experimental findings in MSCs research to clinically applicable therapeutics. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1268-1288, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27152   open full text
• The viral approach to breast cancer immunotherapy.
  Atefeh Arab, Nima Behravan, Atefeh Razazn, Nastaran Barati, Fatemeh Mosaffa, Jessica Nicastro, Roderick Slavcev, Javad Behravan.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- This manuscript discusses the latest findings regarding viral vectors designed for fighting breast cancer Despite years of intensive research, breast cancer remains the leading cause of death in women worldwide. New technologies including oncolytic virus therapies, virus, and phage display are among the most powerful and advanced methods that have emerged in recent years with potential applications in cancer prevention and treatment. Oncolytic virus therapy is an interesting strategy for cancer treatment. Presently, a number of viruses from different virus families are under laboratory and clinical investigation as oncolytic therapeutics. Oncolytic viruses (OVs) have been shown to be able to induce and initiate a systemic antitumor immune response. The possibility of application of a multimodal therapy using a combination of the OV therapy with immune checkpoint inhibitors and cancer antigen vaccination holds a great promise in the future of cancer immunotherapy. Display of immunologic peptides on bacterial viruses (bacteriophages) is also increasingly being considered as a new and strong cancer vaccine delivery strategy. In phage display immunotherapy, a peptide or protein antigen is presented by genetic fusions to the phage coat proteins, and the phage construct formulation acts as a protective or preventive vaccine against cancer. In our laboratory, we have recently tested a few peptides (E75, AE37, and GP2) derived from HER2/neu proto‐oncogene as vaccine delivery modalities for the treatment of TUBO breast cancer xenograft tumors of BALB/c mice. Here, in this paper, we discuss the latest advancements in the applications of OVs and bacterial viruses display systems as new and advanced modalities in cancer immune therapeutics. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1257-1267, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27150   open full text
• MicroRNA‐10b expression predicts long‐term survival in patients with solid tumor.
  Yi Zhang, Li‐Juan Wang, He‐Quan Yang, Rong Wang, Hua‐Jun Wu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 Abstract Background Numerous studies have evaluated the significance of the microRNA‐10b (miR‐10b) in the development and progression of many cancers. Their findings revealed that increased expression of miR‐10b is associated with unfavorable prognosis in patients with cancer. Results A total of 1,834 patients from 19 studies were included in this study. A significantly shorter overall survival was observed in patients with increased expression of miR‐10b (hazard ratio [HR] = 1.99, 95% confidence interval [CI]: 1.51–2.61). Statistical significance was also observed in subgroup meta‐analysis stratified by the cancer type, cutoff value, analysis type, and sample size. Also, patients with a high expression level of miR‐10b had a poorer disease‐free survival rate (HR = 1.18, 95% CI: 1.05–1.33). In addition, the pooled odds ratios (ORs) showed that increased miR‐10b was also associated with positive lymph node metastasis (OR = 2.09, 95% CI: 1.45–3.03), distant metastasis (OR = 2.40, 95% CI: 1.57–3.67), tumor size (OR = 3.86, 95% CI: 2.25–6.64), and poor clinical stage (OR = 5.02, 95% CI: 3.37–7.47). Materials and Methods A systematic literature search was conducted on a number of electronic databases, including PubMed, Embase, Web of Science, China National Knowledge Infrastructure, Springer, Google Scholar, and Gene expression omnibus. We retrieved the relevant articles to examine the association between the miR‐10b expression levels and patients’ prognosis. The meta‐analysis was conducted using the RevMan 5.2 software and Stata SE12.0 software. Conclusions High miR‐10b expression was correlated with poor clinical outcome, which indicated the potential clinical use of miR‐10b as a molecular biomarker for cancer, particularly in assessing prognosis for patients with cancers. Further studies should be performed to verify the clinical utility of miR‐10b in human solid tumors. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1248-1256, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27138   open full text
• Therapeutic potency of Wnt signaling antagonists in the pathogenesis of prostate cancer, current status and perspectives.
  Mehran Pashirzad, Mojtaba Shafiee, Majid Khazaei, Hamid Fiuji, Mikhail Ryzhikov, Saman Soleimanpour, AmirReza Hesari, Amir Avan, Seyed Mahdi Hassanian.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Suppression of Wnt canonical or noncanonical signaling pathways via Wnt biological or pharmacological antagonists is a potentially novel therapeutic approach for patients with prostate cancer. This review summarizes the role of Wnt signaling inhibitors in the pathogenesis of prostate cancer for a better understanding and hence a better management of this disease. Abstract Prostate cancer is a major cause of cancer‐related death in males. Wnt/β‐catenin signaling plays a critical role in the pathogenesis of this disease by regulating angiogenesis, drug resistance, cell proliferation, and apoptosis. Suppression of Wnt canonical or noncanonical signaling pathways via Wnt biological or pharmacological antagonists is a potentially novel therapeutic approach for patients with prostate cancer. This review summarizes the role of Wnt signaling inhibitors in the pathogenesis of prostate cancer for a better understanding and hence a better management of this disease. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1237-1247, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27137   open full text
• The prognostic and therapeutic values of long noncoding RNA PANDAR in colorectal cancer.
  Mahdi Rivandi, Alireza Pasdar, Leila Hamzezadeh, Amir Tajbakhsh, Sima Seifi, Mehrdad Moetamani‐Ahmadi, Gordon A. Ferns, Amir Avan.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Schematic representation of mechanisms of PANDAR in colorectal cancer. PANDAR could promote metastasis via EMT pathway on the other hand, it could regulate cell proliferation. PANDAR inhibits apoptosis by bax‐caspase 3 pathway. Abstract Long noncoding RNAs (lncRNAs) consist of 200 nucleotide sequences that play essential roles in different processes, including cell proliferation, and differentiation. There is evidence showing that the dysregulation of lncRNAs promoter of CDKN1A antisense DNA damage‐activated RNA (PANDAR) leads to the development and progression in several cancers including colorectal cancer, via p53‐dependent manner. This suggests that these lncRNAs may be of value as prognostic indices and a therapeutic target, as a high expression of lncRNAs PANDAR is associated with poor prognosis. Furthermore, modulating lncRNAs PANDAR has been reported to induce apoptosis and inhibit the tumor growth through modulation of cell cycle and epithelial‐mesenchymal transition (EMT) pathway. The aim of the current review was to provide an overview of the prognostic and therapeutic values of lncRNAs PANDAR in colorectal cancer - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1230-1236, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27136   open full text
• Stem cell‐based therapy for Parkinson’s disease with a focus on human endometrium‐derived mesenchymal stem cells.
  Saeid Bagheri‐Mohammadi, Mohammad Karimian, Behrang Alani, Javad Verdi, Rana Moradian Tehrani, Mahdi Noureddini.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Parkinson’s disease (PD) as an increasing clinical syndrome is a multifunctional impairment. Till now, there is a lack of eligible treatment for PD, and stem cells therapy recently has been considered for PD treatment. Human stem cell technology especially human endometrium‐derived stem cells (HEDSCs) have made advancement as a therapeutic source for PD. Abstract Parkinson’s disease (PD) as an increasing clinical syndrome is a multifunctional impairment with systemic involvement. At present, therapeutic approaches such as l‐3,4‐dihydroxy‐phenylalanine replacement therapy, dopaminergic agonist administration, and neurosurgical treatment intend to relieve PD symptoms which are palliative and incompetent in counteracting PD progression. These mentioned therapies have not been able to replace the lost cells and they could not effectively slow down the relentless neurodegenerative process. Till now, there is a lack of eligible treatment for PD, and stem cells therapy recently has been considered for PD treatment. In this review, we demonstrate how human stem cell technology especially human endometrium‐derived stem cells have made advancement as a therapeutic source for PD compared with other treatments. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1326-1335, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27182   open full text
• Heat shock protein 70: A promising therapeutic target for myocardial ischemia–reperfusion injury.
  Yan‐Jun Song, Chong‐Bin Zhong, Xian‐Bao Wang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- During myocardial ischemia–reperfusion (I/R), HSP70 is overexpressed and exerts numerous cardioprotective effects against I/R injury. In this review, we highlight these protective effects of HSP70, elucidate the mechanisms involved, and provide several HSP70‐driven therapeutic potentials against myocardial I/R injury. In summary, it is considered that HSP70 is a promising therapeutic target against myocardial I/R injury. Acute myocardial infarction is a major cause of death worldwide. The most important therapy for limiting ischemic injury and infarct size is timely and efficient myocardial reperfusion treatment, which may instead induce cardiomyocyte necrosis due to myocardial ischemia–reperfusion (I/R) injury. Heat shock protein 70 (HSP70), a stress‐inducible protein, is overexpressed during myocardial I/R. The induced HSP70 is shown to regulate several intracellular proteins (e.g., transcription factors, enzymes, and apoptosis‐related proteins) and signaling pathways (e.g., c‐Jun N‐terminal kinase pathway and extracellular‐signal‐regulated kinase 1/2 pathway), forming a complicated network that contributes to reducing reactive oxygen species accumulation, improving calcium homeostasis, inhibiting cellular apoptosis, thereby enhancing the stress adaption of myocardium to I/R injury. In addition, the extracellular HSP70, which is released from injured cardiomyocytes during I/R, acts as a proinflammatory mediator that results in cell death, while the intracellular HSP70 exerts antiinflammatory effects by suppressing proinflammatory signaling pathways. Notably, HSP70 is induced and contributes to the cardioprotection in several types of preconditioning and postconditioning. Meanwhile, it is shown that the cardioprotective effectiveness of preconditioning‐induced HSP70 (e.g., hyperthermia preconditioning‐induced HSP70) can be impaired by certain pathological conditions, such as hyperlipidemia and hyperglycemia. Thus, we highlight the widespread cardioprotective involvement of HSP70 in preconditioning and postconditioning and elucidate how HSP70‐mediated cardioprotection is impaired in these pathological conditions. Furthermore, several therapeutic potentials of HSP70 against myocardial I/R injury and potential directions for future studies are also provided in this review. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1190-1207, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27110   open full text
• Therapeutic potential of A2 adenosine receptor pharmacological regulators in the treatment of cardiovascular diseases, recent progress, and prospective.
  Amirhossein Bahreyni, Amir Avan, Mohammad Shabani, Mikhail Ryzhikov, Hamid Fiuji, Saman Soleimanpour, Majid Khazaei, Seyed Mahdi Hassanian.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Regulation of A2 adenosine receptor signaling via specific and novel pharmacological regulators is a potentially novel therapeutic approach for a better understanding and hence a better management of cardiovascular diseases (CVDs). This review summarizes the role of pharmacological A2 adenosine receptor regulators in the pathogenesis of CVDs. Abstract Adenosine and its analogs are of particular interest as potential therapeutic agents for treatment of cardiovascular diseases (CVDs). A2 adenosine receptor subtypes (A2a and A2b) are extensively expressed in cardiovascular system, and modulation of these receptors using A2 adenosine receptor agonists or antagonists regulates heart rate, blood pressure, heart rate variability, and cardiovascular toxicity during both normoxia and hypoxia conditions. Regulation of A2 adenosine receptor signaling via specific and novel pharmacological regulators is a potentially novel therapeutic approach for a better understanding and hence a better management of CVDs. This review summarizes the role of pharmacological A2 adenosine receptor regulators in the pathogenesis of CVDs. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1295-1299, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27161   open full text
• The clinical importance of CD4+CD7− in human diseases.
  Saeed Mohammadian Haftcheshmeh, Amir Tajbakhsh, Mohaddese Kazemi, Seyed‐Alireza Esmaeili, Fatemeh Mardani, Mostafa Fazeli, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this review, we first focus on the clinical importance of CD4+CD7− T cells in human disease and also explain the possible function of these cells. Secondly, we also investigate if the CD4 +CD7− T cells are resistant to apoptosis. The CD7 antigen is a member of the immunoglobulin superfamily that expresses on the surface of all thymocytes, a majority of mature T cells, and also natural killer cells. Interestingly, under physiological and different pathological conditions, the loss of CD7 antigen occurred in the subset of CD4+ memory T cells. Various functions have been proposed for CD7, including its role in the activation and intercellular adhesiveness of T cells. Several studies indicate that the number of CD4+CD7− T cells increases in diseases such as chronic inflammation and T‐cell malignancies, these being skin inflammatory lesions. Therefore, this can be useful for the diagnosis of cancer cells, especially with reference to blood origin, treatment monitoring, and establishment of new therapies. Therefore, a comprehensive review could be useful to increase our knowledge about the clinical importance of these cells in human disease. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1179-1189, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27099   open full text
• Evidence of curcumin and curcumin analogue effects in skin diseases: A narrative review.
  Yunes Panahi, Omid Fazlolahzadeh, Stephen L. Atkin, Muhammed Majeed, Alexandra E. Butler, Thomas P. Johnston, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Curcumin, a natural polyphenolic and yellow pigment obtained from the spice turmeric, has strong antioxidative, anti‐inflammatory, and antibacterial properties; due to these properties, curcumin has been used as a remedy for the prevention and treatment of skin aging and disorders such as psoriasis, infection, acne, skin inflammation, and skin cancer and curcumin has protective effects against skin damage caused by chronic ultraviolet B radiation. One of the challenges in maximizing the therapeutic potential of curcumin is its low bioavailability, limited aqueous solubility, and chemical instability; in this regard, the present review is focused on recent studies concerning the use of curcumin for the treatment of skin diseases, as well as offering new and efficient strategies to optimize its pharmacokinetic profile and increase its bioavailability. Curcumin, a natural polyphenolic and yellow pigment obtained from the spice turmeric, has strong antioxidative, anti‐inflammatory, and antibacterial properties. Due to these properties, curcumin has been used as a remedy for the prevention and treatment of skin aging and disorders such as psoriasis, infection, acne, skin inflammation, and skin cancer. Curcumin has protective effects against skin damage caused by chronic ultraviolet B radiation. One of the challenges in maximizing the therapeutic potential of curcumin is its low bioavailability, limited aqueous solubility, and chemical instability. In this regard, the present review is focused on recent studies concerning the use of curcumin for the treatment of skin diseases, as well as offering new and efficient strategies to optimize its pharmacokinetic profile and increase its bioavailability. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1165-1178, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27096   open full text
• Exercise ameliorates high‐fat diet‐induced impairment of differentiation of adipose‐derived stem cells into neuron‐like cells in rats.
  Hisashi Kato, Hidemasa Minamizato, Hideki Ohno, Yoshinobu Ohira, Tetsuya Izawa.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- High‐fat diet (HFD) impaired the differentiation potential of adipose‐derived stem cells (ADSCs) into neuron‐like cells, which was accompanied by increases in apoptotic activity and oxidative stress. Importantly, exercise training ameliorated the HFD‐induced impairment of neurogenesis in ADSCs. Thus, the adipose tissue microenvironment could influence the differentiation potential of ADSCs, a source of autologous stem cell therapy. Adipose‐derived stem cells (ADSCs) can differentiate into neurons under particular conditions. It remains largely unknown whether this differentiation potential is affected by physical conditions such as obesity, which modulates the functions of adipose tissue. In this study, we determined the impact of either a 9‐week high‐fat diet (60% fat; HFD) or 9‐week exercise training on the differentiation potential of ADSCs into neuron‐like cells in male Wistar rats. Rats were randomly assigned to a normal diet‐fed (ND‐SED) group, HFD‐fed (HFD‐SED) group, or exercise‐trained HFD‐fed group (HFD‐EX). After a 9‐week intervention, ADSCs from all groups differentiated into neuron‐like cells. Expression of neuronal marker proteins (nestin, βIII‐tubulin, and microtubule‐associated protein 2 [MAP2]) and the average length of cell neurites were lower in cells from HFD‐SED rats than in other groups. Instead, protein expression of COX IV and Cyt‐c, the Bax/Bcl‐2 and LC3‐II/I ratio, and the malondialdehyde level in culture medium were higher in cells from HFD‐SED rats. No significant difference between ND‐SED and HFD‐EX rats was observed, except for the average length of cell neurites in MAP2. Thus, HFD impaired the differentiation potential of ADSCs into neuron‐like cells, which was accompanied by increases in apoptotic activity and oxidative stress. Importantly, exercise training ameliorated the HFD‐induced impairment of neurogenesis in ADSCs. The adipose tissue microenvironment could influence the differentiation potential of ADSCs, a source of autologous stem cell therapy. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1452-1460, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26957   open full text
• Wogonoside impedes the progression of acute myeloid leukemia through inhibiting bone marrow angiogenesis.
  Binyan Lin, Kai Zhao, Dawei Yang, Dongsheng Bai, Yan Liao, Yuxin Zhou, Zhou Yu, Xiaoxuan Yu, Qinglong Guo, Na Lu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Wogonoside inhibits bone marrow angiogenesis in acute myeloid leukemia (AML) patients. Wogonoside inhibits angiogenesis via JAK‐STAT3 signaling. Wogonoside would be a promising drug for relapsed and resistant AML patients. Decreasing bone marrow (BM) microvessel density and circulating angiogenic cytokine levels are promising strategies for the treatment of relapsed and resistant acute myeloid leukemia (AML). Previous studies have reported that wogonoside could inhibit the progression of AML and suppress angiogenesis in a solid tumor, but the correlation of these two effects was ignored. In this research, we determined whether wogonoside could inhibit angiogenesis in this hematologic malignancy. We found that wogonoside could inhibit tumor growth and progression, and prolong the survival of nude mice inoculated with U937/MDR. Besides, reducing BM angiogenesis might cause therapeutic effect against resistant AML. Therefore, coculture between AML cells and BM stromal cells was established to imitate their crosstalk. Then, the effect of wogonoside on BM angiogenesis was tested in vitro and in vivo. We found that wogonoside could suppress microvessel formation in the chicken chorioallantoic membrane assay model and matrigel plug assay. The mechanism research revealed that wogonoside could block the JAK2‐STAT3 pathway in AML cells and stromal cells to break their positive feedback. We detected several cytokines related to AML or angiogenesis and found that secreted interleukin‐8 was a significant angiogenic cytokine to induce BM angiogenesis. These findings supported that new diagnostics and promising treatment strategies could be developed in relapsed and resistant AML patients. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1913-1924, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27067   open full text
• Prolonged lipopolysaccharide exposure induces transient immunosuppression in BV2 microglia.
  Krishna Sundar Twayana, Namrata Chaudhari, Palaniyandi Ravanan.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In vitro endotoxin treatment on BV2 microglia cells has similar results, that is, cells become hyporesponsive to stimuli, even after repeated exposure to lipopolysaccharide for months. Immunosuppressed cells reversing back to normal immunoresponsive proves plasticity of these central nervous system macrophages. Nature has enabled them to have such plastic nature to be adapted themselves first and continue the immunosurveillance to protect neuronal cells that are comparatively fragile and vulnerable to inflammatory stress. And endogenously high inhibitor of apoptotic proteins expression enables them to cope up with continual exposure to stimuli without compromising the viability. Continuous pre‐exposure of immune cells to low level of inflammatory stimuli makes them hyporesponsive to subsequent exposure. This pathophysiological adaptation; known as endotoxin tolerance is a general paradigm behind several disease pathogenesis. Current study deals with this immunosuppression with respect to BV2 microglia. We attempted to investigate their immune response under prolonged endotoxin exposure and monitor the same upon withdrawal of the stimuli. BV2 microglia cells were maintained under continual exposure of lipopolysaccharide (LPS) for weeks with regular passage after 72 hr (prolonged LPS exposed cells [PLECs]). PLECs were found to be immunosuppressed with diminished expression of proinflammatory cytokines (IL6, IL1β, TNF‐α, and iNOS) and production of nitric oxide, as compared to once LPS exposed cells. Upon remaintenance of cells in normal media without LPS exposure (LPS withdrawal cells [LWCs]), the induced immunosuppression reversed and cells started responding to inflammatory stimuli; revealed by significant expression of proinflammatory cytokines. LWCs showed functional similarities to never LPS exposed cells (NLECs) in phagocytosis activity and their response to anti‐inflammatory agents like dexamethasone. Despite their immunoresponsiveness, PLECs were inflamed and showed higher autophagy rate than NLECs. Additionally, we investigated the role of inhibitor of apoptotic proteins (IAPs) in PLECs to understand whether IAPs aids in the survival of microglial cells under stress conditions. Our results revealed that cIAP1 and cIAP2 are induced in PLECs which might play a role in retaining the viability. Furthermore, antagonism of IAPs has significantly induced cell death in PLECs suggesting the role of IAPs in microglial survival under stress condition. Conclusively, our data suggest that continuous exposure of BV2 microglia cells to LPS results in transient immunosuppression and indicates the involvement of IAPs in retaining their viability under inflammatory stress. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1889-1903, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27064   open full text
• Intratumor lactate levels reflect HER2 addiction status in HER2‐positive breast cancer.
  Lorenzo Castagnoli, Egidio Iorio, Matteo Dugo, Ada Koschorke, Simona Faraci, Rossella Canese, Patrizia Casalini, Patrizia Nanni, Claudio Vernieri, Massimo Di Nicola, Daniele Morelli, Elda Tagliabue, Serenella M. Pupa.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- The purpose of this study was to identify a reliable, noninvasive, and novel biomarker associated with HER2 addiction that reflects the susceptibility to HER2‐specific therapy. Our results provide experimental and relevant support for the preclinical and clinical association between HER2 addiction/trastuzumab susceptibility and intratumor levels of lactate, a key metabolic player in cancer progression that is upregulated early in the development of malignancies. Through molecular and metabolic analyses, we found a statistically significant association among HER2 transcript levels, intratumor lactate levels and response to therapy, thus paving the way for the utilization of Magnetic Resonance Imaging and Magnetic Resonance Spectroscopy as a powerful and independent tool to better tailor anti‐HER2 therapies and improve the outcomes of all patients with HER2‐positive BC. Despite different molecular tumor profiles indicate that human epidermal growth factor receptor 2 (HER2) messenger RNA (mRNA) levels mirror HER2 addiction and trastuzumab benefit in HER2‐positive breast cancer (BC), the identification of noninvasive clinical predictors of trastuzumab sensitivity remains an unmet clinical need. In the current study, we investigated whether intratumor lactate levels reflect HER2 addiction and, in turn, trastuzumab susceptibility. Accordingly, the gene expression profiles of transgenic murine BC cell lines expressing the human d16HER2 variant (HER2‐addicted) or human full‐length HER2 (WTHER2; HER2‐nonaddicted) revealed a significant enrichment of glycolysis‐related gene pathways in HER2‐addicted cells. We studied the metabolic content of 22 human HER2‐positive BC by quantitative nuclear magnetic resonance spectroscopy and found that those cases with higher lactate levels were characterized by higher HER2 transcript levels. Moreover, gene expression analyses of HER2‐positive BC samples from a TCGA data set revealed a significant enrichment in glycolysis‐related pathways in high/HER2‐addicted tumors. These data were confirmed by metabolic analyses of human HER2‐positive BC cell lines with high or low HER2 transcript levels, which revealed significantly more active glycolytic metabolism in high HER2 transcript than in low HER2 transcript cells. Overall, our results provide evidence for noninvasive intratumor lactate detection as a potential metabolic biomarker of HER2 addiction and trastuzumab response suggesting the possibility to use in vivo imaging to assess lactate levels and, in turn, select HER2‐positive BC patients who are more likely to benefit from anti‐HER2 treatments. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1768-1779, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27049   open full text
• MicroRNA‐150 contributes to ischemic stroke through its effect on cerebral cortical neuron survival and function by inhibiting ERK1/2 axis via Mal.
  Hui Lv, Jie Li, Yu‐Qin Che.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- The key findings of this study support the notion that miR‐150 under‐expression‐mediated direct promotion of Mal protects CNN functions through the activation of ERK1/2 axis, and underscore the concept that miR‐150 may represent a novel pharmacological target for ischemic stroke intervention. Ischemic stroke, caused by the blockage of blood supply, is a major cause of death worldwide. For identifying potential candidates, we explored the effects microRNA‐150 (miR‐150) has on ischemic stroke and its underlying mechanism by developing a stable middle cerebral artery occlusion (MCAO) rat model. Gene expression microarray analysis was performed to screen differentially expressed genes associated with MCAO. We evaluated the expression of miR‐150 and Mal and the status of ERK1/2 axis in the brain tissues of MCAO rats. Then the cerebral cortical neurons (CCNs) were obtained and introduced with elevated or suppressed miR‐150 or silenced Mal to validate regulatory mechanisms for miR‐150 governing Mal in vitro. The relationship between miR‐150 and Mal was verified by dual luciferase reporter gene assay. Besides, cell growth and apoptosis of CCNs were detected by means of MTT assay and flow cytometry analyses. We identified Mal as a downregulated gene in MCAO, based on the microarray data of GSE16561. MiR‐150 was over‐expressed and negatively targeted Mal in the brain tissues obtained from MCAO rats and their CCNs. Increasing miR‐150 blocked the ERK1/2 axis, resulting in an inhibited cell growth of CNNs but an enhanced apoptosis. Furthermore, MiR‐150 inhibition was observed to have effects on CNNs as opposed to those inhibited by miR‐150 promotion. The key findings of this study support the notion that miR‐150 under‐expression‐mediated direct promotion of Mal protects CNN functions through the activation of the ERK1/2 axis, and underscore the concept that miR‐150 may represent a novel pharmacological target for ischemic stroke intervention. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1477-1490, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26960   open full text
• Hippo pathway functions as a downstream effector of AKT signaling to regulate the activation of primordial follicles in mice.
  Liao‐Liao Hu, Tie Su, Rui‐Chen Luo, Yue‐Hui Zheng, Jian Huang, Zhi‐Sheng Zhong, Jing Nie, Li‐Ping Zheng.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Our study provided direct evidence to highlight the role of Hippo signaling in regulating follicle development and indicated that AKT functions as an upstream regulator of Hippo pathway in primordial follicle activation. Our results help to further clarify the molecular mechanism of the maintenance of primordial follicles pool and suggest a new therapeutic strategy for prevention and treatment of female infertility. Clarifying the molecular mechanisms by which primordial follicles are initiated is crucial for the prevention and treatment of female infertility and ovarian dysfunction. The Hippo pathway has been proven to have a spatiotemporal correlation with the size of the primordial follicle pool in mice in our previous work. But the role and underlying mechanisms of the Hippo pathway in primordial follicle activation remain unclear. Here, the localization and expression of the core components were examined in primordial follicles before and after activation. And the effects of the Hippo pathway on primordial follicle activation were determined by genetically manipulating yes‐associated protein 1 (Yap1), the key transcriptional effector. Furthermore, an AKT specific inhibitor (MK2206) was added to determine the interaction between the Hippo pathway and AKT, an important signaling regulator of ovarian function. Results showed that the core components of the Hippo pathway were localized in both primordial and primary follicles and the expression levels of them changed significantly during the initiation of primordial follicles. Yap1 knockdown suppressed primordial follicle activation, while its overexpression led to the opposite trend. MK2206 downregulated the ratio of P‐MST/MST1 and upregulated the ratio of P‐YAP1/YAP1 significantly, whereas Yap1‐treatment had no influence on AKT. In addition, YAP1 upregulation partially rescued the suppression of the primordial follicle activation induced by MK2206. Our findings revealed that the Hippo‐YAP1 regulates primordial follicular activation, which is mediated by AKT signaling in mice, thus providing direct and new evidence to highlight the role of Hippo signaling in regulating ovarian follicles development. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1578-1587, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27024   open full text
• Caveolin enhances hepatocellular carcinoma cell metabolism, migration, and invasion in vitro via a hexokinase 2‐dependent mechanism.
  Fang Chai, Yan Li, Keyi Liu, Qiang Li, Hongzhi Sun.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Caveolin‐1 (CAV‐1) increased hexokinase 2 (HK2) expression. CAV‐1 enhances glycolysis, invasion, and metastasis in hepatocellular carcinoma cells via a HK2‐dependent mechanism. The development and progression of hepatocellular carcinoma (HCC) have been associated with abnormal cellular metabolism. Gene Expression Profiling Interactive Analysis RNA sequencing data revealed caveolin‐1 (CAV‐1) and hexokinase 2 (HK2) messenger RNA (mRNA) were significantly upregulated in human HCC compared with normal tissues, and high HK2 expression was associated with significantly poorer overall survival in HCC ( p < 0.05). CAV‐1 and HK2 mRNA and protein expression were upregulated and positively correlated in 42 fresh human HCC tissues compared with tumor‐adjacent normal tissues. Overexpression of CAV‐1 or HK2 in SMMC‐7721 and HepG2 HCC cells enhanced glucose and lactate metabolism and increased cell migration and invasion in transwell assays; knocking down CAV‐1 or HK2 had the opposite effects. Overexpression of CAV‐1 increased HK2 expression; overexpression of HK2 did not affect CAV‐1 expression. Knocking down HK2 partially reversed the ability of CAV‐1 to promote cellular metabolism, invasion, and migration in HCC, indicating CAV‐1 enhances glycolysis, invasion, and metastasis in HCC cells via HK2‐dependent mechanism. Further studies of the function and relationship between CAV‐1 or HK2 expression are warranted to explore the potential of these proteins as metabolic targets for the treatment of HCC. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1937-1946, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27074   open full text
• Mutual regulation between IGF‐1R and IGFBP‐3 in human corneal epithelial cells.
  Rossella Titone, Meifang Zhu, Danielle M. Robertson.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Here we report that the insulin‐like growth factor binding protein‐3 (IGFBP‐3) mediates nuclear translocation of insulin‐like growth factor type 1 receptor (IGF‐1R) in response to stress. This occurs via SUMOylation by SUMO 2/3. This is the first study to show a direct relationship between IGF‐1R and IGFBP‐3 in the maintenance of corneal epithelial homeostasis. The insulin‐like growth factor type 1 receptor (IGF‐1R) is part of the receptor tyrosine kinase superfamily. The activation of IGF‐1R regulates several key signaling pathways responsible for maintaining cellular homeostasis, including survival, growth, and proliferation. In addition to mediating signal transduction at the plasma membrane, in serum‐based models, IGF‐1R undergoes SUMOylation by SUMO 1 and translocates to the nucleus in response to IGF‐1. In corneal epithelial cells grown in serum‐free culture, however, IGF‐1R has been shown to accumulate in the nucleus independent of IGF‐1. In this study, we report that the insulin‐like growth factor binding protein‐3 (IGFBP‐3) mediates nuclear translocation of IGF‐1R in response to growth factor withdrawal. This occurs via SUMOylation by SUMO 2/3. Further, IGF‐1R and IGFBP‐3 undergo reciprocal regulation independent of PI3k/Akt signaling. Thus, under healthy growth conditions, IGFBP‐3 functions as a gatekeeper to arrest the cell cycle in G0/G1, but does not alter mitochondrial respiration in cultured cells. When stressed, IGFBP‐3 functions as a caretaker to maintain levels of IGF‐1R in the nucleus. These results demonstrate mutual regulation between IGF‐1R and IGFBP‐3 to maintain cell survival under stress. This is the first study to show a direct relationship between IGF‐1R and IGFBP‐3 in the maintenance of corneal epithelial homeostasis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1426-1441, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26948   open full text
• Norepinephrine stimulation downregulates the β2‐adrenergic receptor–nitric oxide pathway in human pulmonary artery endothelial cells.
  Wande Yu, Yue Gu, PeiP Chen, Jie Luo, Pengfei Liu, Yuelin Chao, Shao‐Liang Chen, Hang Zhang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- These data provide a novel mechanism for norepinephrine (NE)‐decreased endothelium‐derived nitric oxide and NE‐induced human pulmonary artery endothelial cells proliferation that leads to pulmonary hypertension associated with left heart disease (PH‐LHD), suggesting a potential therapeutic target for PH‐LHD. Background Norepinephrine (NE)‐mediated vasoconstriction plays an important role in pulmonary hypertension associated with left heart disease (PH‐LHD). However, the role of NE‐mediated endothelial cell dysfunction in the pathogenesis of PH‐LHD remains to be elucidated. Methods and Results An enzyme‐linked immunosorbent assay showed that the NE concentration in the plasma of patients with PH‐LHD was higher and the nitrate–nitrite concentration was lower than those in the control group. NE treatment decreased phospho‐Ser633‐eNOS and β2‐adrenergic receptor (β2‐AR) levels in the membrane of human pulmonary artery endothelial cells (HPAECs) analysed by western blot analysis. Consistently, fluorescence microscopy and flow cytometry showed that nitric oxide (NO) production was also decreased in HPAECs. Coimmunoprecipitation confirmed a direct interaction between β2‐AR and endothelial NO synthase (eNOS). Overexpression of β2‐AR attenuated the decline in phospho‐Ser633‐eNOS and NO production. Additionally, the expression of phospho‐Ser633‐eNOS and β2‐AR was decreased in human pulmonary artery endothelium. Finally, our results indicate that NE stimulated HPAEC proliferation, which was blocked by protein kinase A inhibitor or protein kinase B (PKB–AKT) inhibitor. Conclusions These data provide a novel mechanism for NE‐decreased endothelium‐derived NO and NE‐induced HPAEC proliferation that leads to PH‐LHD, suggesting a potential therapeutic target for PH‐LHD. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1842-1850, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27057   open full text
• Carvacrol induces mitochondria‐mediated apoptosis via disruption of calcium homeostasis in human choriocarcinoma cells.
  Whasun Lim, Jiyeon Ham, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this study, we demonstrated that carvacrol exertes antiproliferative effects through suppression of phosphoinositide 3‐kinase–protein kinase B and mitogen‐activated protein kinase signaling, generating oxidative stress and altering calcium homeostasis mediated by mitochondrial dysfunction in human choriocarcinoma JAR and JEG3 cells. Therefore, carvacrol has a possibility as a novel therapeutic agent or supplement for the treatment of choriocarcinoma patients. Carvacrol is a monoterpenoid phenol present in the oils of various plants including Origanum vulgare (oregano) or Origanum majorana (marjoram). For a long time, it has been used as spice in foods because of its antimicrobial properties. Additionally, it appears to have anticancer effects against some cancer but this has not been well studied. Therefore, we conducted various assays to confirm the effects of carvacrol on choriocarcinoma cell lines (JAR and JEG3). Our results indicate that carvacrol has antiproliferative properties and induces apoptosis, resulting in increased expression of proapoptotic proteins. Additionally, carvacrol disrupted the mitochondrial membrane potential and induced calcium ion overload in the mitochondrial matrix in both JAR and JEG3 cells. Furthermore, carvacrol generated oxidative stress and lipid peroxidation in both JAR and JEG3 cells. Moreover, carvacrol‐suppressed phosphoinositide 3‐kinase–protein kinase B and extracellular signal–regulated kinase 1/2 mitogen‐activated protein kinase (MAPK) signal transduction whereas expression of phosphor‐P38 and c‐Jun N‐terminal kinase MAPK was increased. Together, our results indicate that carvacrol may be a possible new therapeutic agent or supplement for the control of human choriocarcinomas. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1803-1815, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27054   open full text
• The predictive value of a preoperative systemic immune‐inflammation index and prognostic nutritional index in patients with esophageal squamous cell carcinoma.
  Hongdian Zhang, Xiaobin Shang, Peng Ren, Lei Gong, Ashique Ahmed, Zhao Ma, Rong Ma, Xianxian Wu, Xiangming Xiao, Hongjing Jiang, Peng Tang, Zhentao Yu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- (a) The preoperative high systemic immune‐inflammation index (SII) and low prognostic nutritional index (PNI) are powerful indicators of aggressive biology and poor prognosis for patients with esophageal squamous cell carcinoma (ESCC). (b) Combination of SII and PNI (coSII–PNI) clearly classified ESCC patients into three independent groups before surgery. (c) The coSII–PNI can enhance the accuracy of prognosis. Growing evidence indicates that systemic inflammation response and malnutrition status are correlated with survival in certain types of solid tumors. The aim of this study is to evaluate the association between the systemic immune‐inflammation index (SII) and prognostic nutritional index (PNI) and overall survival (OS) in patients with esophageal squamous cell carcinoma (ESCC) after esophagectomy. A consecutive series of 655 patients with resected ESCC who underwent esophagectomy were enrolled in the retrospective study. The preoperative SII was defined as platelet × neutrophil/lymphocyte counts. The PNI was calculated as albumin concentration (g/L) + 5 × total lymphocyte count (109/L). The optimal cut‐off values of SII, neutrophil‐to‐lymphocyte ratio (NLR), platelet‐to‐lymphocyte ratio (PLR), and PNI were determined by receiver operating characteristic analysis. Survival analysis was performed using the Kaplan–Meier method with a log‐rank test, followed by a multivariate Cox proportional hazards model. A high SII was significantly related to tumor size, histological type, invasion depth, and TNM stage (p < 0.05). A low PNI was significantly associated with age, tumor size, invasion depth, lymph node metastasis, and TNM stage (p < 0.05). Univariate analysis revealed that age, smoking history, tumor size, invasion depth, lymph node metastasis, SII, NLR, PLR, and PNI were predictors of OS (p < 0.05). Multivariate analysis identified age (p = 0.041), tumor size (p = 0.016), invasion depth (p < 0.001), lymph node metastasis (p < 0.001), SII (p = 0.033), and PNI (p = 0.022) as independent prognostic factors correlated with OS. There was a significant inverse relationship between the SII and PNI (r = 0.309; p < 0.001). The predictive value increased when the SII and PNI were considered in combination. Our results demonstrate that the preoperative high SII and low PNI are powerful indicators of aggressive biology and poor prognosis for patients with ESCC. The combination of SII and PNI can enhance the accuracy of prognosis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1794-1802, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27052   open full text
• Irisin promotes cardiac progenitor cell‐induced myocardial repair and functional improvement in infarcted heart.
  Yu Tina Zhao, Jianguo Wang, Naohiro Yano, Ling X. Zhang, Hao Wang, Shouyan Zhang, Gangjian Qin, Patrycja M. Dubielecka, Shougang Zhuang, Paul Y. Liu, Y. Eugene Chin, Ting C. Zhao.
  Journal of Cellular Physiology. November 20, 2018
  --- - "\nThese results indicate that Irisin promoted Nkx2.5+ cardiac progenitor cell (CPC)‐induced cardiac regeneration and functional improvement and that Irisin serves as a novel therapeutic approach for stem cells in cardiac repair. MI: myocardial infarction. \n\n\n\n\n\nIrisin, a newly identified hormone and cardiokine, is critical for modulating body metabolism. New evidence indicates that irisin protects the heart against myocardial ischemic injury. However, whether irisin enhances cardiac progenitor cell (CPC)‐induced cardiac repair remains unknown. This study examines the effect of irisin on CPC‐induced cardiac repair when these cells are introduced into the infarcted myocardium. Nkx2.5+ CPC stable cells were isolated from mouse embryonic stem cells. Nkx2.5\n+ CPCs (0.5 × 10\n6) were reintroduced into the infarcted myocardium using PEGlylated fibrin delivery. The mouse myocardial infarction model was created by permanent ligation of the left anterior descending (LAD) artery. Nkx2.5\n+ CPCs were pretreated with irisin at a concentration of 5 ng/ml in vitro for 24 hr before transplantation. Myocardial functions were evaluated by echocardiographic measurement. Eight weeks after engraftment, Nkx2.5\n+ CPCs improved ventricular function as evident by an increase in ejection fraction and fractional shortening. These findings are concomitant with the suppression of cardiac hypertrophy and attenuation of myocardial interstitial fibrosis. Transplantation of Nkx2.5\n+ CPCs promoted cardiac regeneration and neovascularization, which were increased with the pretreatment of Nkx2.5\n+ CPCs with irisin. Furthermore, irisin treatment promoted myocyte proliferation as indicated by proliferative markers Ki67 and phosphorylated histone 3 and decreased apoptosis. Additionally, irisin resulted in a marked reduction of histone deacetylase 4 and increased p38 acetylation in cultured CPCs. These results indicate that irisin promoted Nkx2.5\n+ CPC‐induced cardiac regeneration and functional improvement and that irisin serves as a novel therapeutic approach for stem cells in cardiac repair." - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1671-1681, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27037   open full text
• NKCC1 promotes EMT‐like process in GBM via RhoA and Rac1 signaling pathways.
  Haiwen Ma, Tao Li, Zhennan Tao, Long Hai, Luqing Tong, Li Yi, Iruni R. Abeysekera, Peidong Liu, Yang Xie, Jiabo Li, Feng Yuan, Chen Zhang, Yihan Yang, Haolang Ming, Shengping Yu, Xuejun Yang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- There are some highlights in this paper: 1. An ion channel that is critical to tumor invasion is proposed. 2. This Ion cotransporters promoted epithelial‐mesenchymal transition process in glioma. 3. An intermediate mechanism for regulating epithelial–mesenchymal transition is proved. Glioblastoma is the most common and lethal primary intracranial tumor. As the key regulator of tumor cell volume, sodium‐potassium‐chloride cotransporter 1 (NKCC1) expression increases along with the malignancy of the glioma, and NKCC1 has been implicated in glioblastoma invasion. However, little is known about the role of NKCC1 in the epithelial‐mesenchymal transition‐like process in gliomas. We noticed that aberrantly elevated expression of NKCC1 leads to changes in the shape, polarity, and adhesion of cells in glioma. Here, we investigated whether NKCC1 promotes an epithelial–mesenchymal transition (EMT)‐like process in gliomas via the RhoA and Rac1 signaling pathways. Pharmacological inhibition and knockdown of NKCC1 both decrease the expressions of mesenchymal markers, such as N‐cadherin, vimentin, and snail, whereas these treatments increase the expression of the epithelial marker E‐cadherin. These findings indicate that NKCC1 promotes an EMT‐like process in gliomas. The underlying mechanism is the facilitation of the binding of Rac1 and RhoA to GTP by NKCC1, which results in a significant enhancement of the EMT‐like process. Specific inhibition or knockdown of NKCC1 both attenuate activated Rac1 and RhoA, and the pharmacological inhibitions of Rac1 and RhoA both impair the invasion and migration abilities of gliomas. Furthermore, we illustrated that NKCC1 knockdown abolished the dissemination and spread of glioma cells in a nude mouse intracranial model. These findings suggest that elevated NKCC1 activity acts in the regulation of an EMT‐like process in gliomas, and thus provides a novel therapeutic strategy for targeting the invasiveness of gliomas, which might help to inhibit the spread of malignant intracranial tumors. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1630-1642, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27033   open full text
• mTOR inhibitor INK128 attenuates dextran sodium sulfate‐induced colitis by promotion of MDSCs on Treg cell expansion.
  Guoping Shi, Dan Li, Jing Ren, Xiaojing Li, Tingting Wang, Huan Dou, Yayi Hou.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Mammalian target of rapamycin inhibitor INK128 attenuates dextran sodium sulfate–induced colitis by promotion of myeloid‐derived suppressor cells on regulatory T‐cell expansion. Accumulating evidence has shown that mammalian target of rapamycin (mTOR) pathway and myeloid‐derived suppressor cells (MDSCs) are involved in pathogenesis of inflammatory bowel diseases (IBDs). INK128 is a novel mTOR kinase inhibitor in clinical development. However, the exact roles of MDSCs and INK128 in IBD are unclear. Here, we showed that the INK128 treatment enhanced the resistance of mice to dextran sodium sulfate (DSS)–induced colitis and inhibited the differentiation of MDSCs into macrophages. Moreover, interferon (IFN)‐α level was elevated in INK128‐treated colitis mice. When stimulated with IFN‐α in vitro, MDSCs showed a superior immunosuppression activity. Of note, the regulatory T cells (Tregs) increased but Th1 cells decreased in INK128‐treated colitis mice. These results indicate that mTOR inhibitor INK128 attenuates DSS‐induced colitis via Treg expansion promoted by MDSCs. Our work provides a new evidence that INK128 is potential to be a therapeutic drug on DSS‐induced colitis via regulating MDSCs as well as maintaining Treg expansion. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1618-1629, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27032   open full text
• LncRNA AK077216 promotes RANKL‐induced osteoclastogenesis and bone resorption via NFATc1 by inhibition of NIP45.
  Chuan Liu, Zhen Cao, Yun Bai, Ce Dou, Xiaoshan Gong, Mengmeng Liang, Rui Dong, Hongyu Quan, Jianmei Li, Jingjin Dai, Fei Kang, Chunrong Zhao, Shiwu Dong.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Lnc‐AK077216 promotes the expression of NFATc1 via suppressing NIP45 expression, thereby promoting RANKL‐induced osteoclast differentiation and function. Osteoclasts derived from the monocyte/macrophage hematopoietic lineage regulate bone resorption, a process balanced by bone formation in the continual renewal of the skeletal system. As dysfunctions of these cells result in bone metabolic diseases such as osteoporosis and osteopetrosis, the exploration of the mechanisms regulating their differentiation is a priority. A potential mechanism may involve long noncoding RNAs (lncRNAs), which are known to regulate various cell biology activities, including proliferation, differentiation, and apoptosis. The expression of the lncRNA AK077216 (Lnc‐AK077216) is significantly upregulated during osteoclastogenesis identified by microarray and verified by qPCR. Up‐ and downregulation of Lnc‐AK077216, respectively promotes and inhibits osteoclast differentiation, bone resorption, and the expression of related genes on the basis of tartrate‐resistant acid phosphatase staining, qPCR, and western blot results. In addition, Lnc‐AK077216 suppresses NIP45 expression and promotes the expression of NFATc1, an essential transcription factor during osteoclastogenesis. Besides, it was found that the expression of Lnc‐AK077216 and Nfatc1 is upregulated, whereas Nip45 expression is downregulated in bone marrow and spleen tissues of ovariectomized mice. The results suggest that Lnc‐AK077216 regulates NFATc1 expression and promotes osteoclast formation and function, providing a novel mechanism of osteoclastogenesis and a potential biomarker or a new drug target for osteoporosis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1606-1617, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27031   open full text
• MiR‐24‐3p regulates cell proliferation and milk protein synthesis of mammary epithelial cells through menin in dairy cows.
  Cao Qiaoqiao, Honghui Li, Xue Liu, Zhengui Yan, Meng Zhao, Zhongjin Xu, Zhonghua Wang, Kerong Shi.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 MiR‐24‐3p negatively modulates the expression of MEN1/menin in mammary epithelial cells by targeting the 3 'UTR of MEN1. MiR‐24‐3p plays regulatory role of cell proliferation and milk protein synthesis in mammary epithelial cells through cooperatively acting with menin. Abstract MiR‐24‐3p, a broadly conserved, small, noncoding RNA, is abundantly expressed in mammary tissue. However, its regulatory role in this tissue remains poorly understood. It was predicted that miR‐24‐3p targets the 3′ untranslated region (3′‐UTR) of multiple endocrine neoplasia type 1 (MEN1), an important regulatory factor in mammary tissue. The objective of this study was to investigate the function of miR‐24‐3p in mammary cells. Using a luciferase assay in mammary epithelial cells (MAC‐T), miR‐24‐3p was confirmed to target the 3′‐UTR of MEN1. Furthermore, miR‐24‐3p negatively regulated the expression of the MEN1 gene and its encoded protein, menin. miR‐24‐3p enhanced proliferation of MAC‐T by promoting G1/S phase progression. MiR‐24‐3p also regulated the expression of key factors involved in phosphatidylinositol‐3‐kinase/protein kinase B/mammalian target of rapamycin and Janus kinase/signal transducer and activators of transcription signaling pathways, therefore controlling milk protein synthesis in epithelial cells. Thus, miR‐24‐3p appears to act on MAC‐T by targeting MEN1. The expression of miR‐24‐3p was controlled by MEN1/menin, indicating a negative feedback loop between miR‐24‐3p and MEN1/menin. The negatively inhibited expression pattern of miR‐24‐3p and MEN1 was active in mammary tissues at different lactation stages. The feedback mechanism is a new concept to further understand the lactation cycle of mammary glands and can possibly to be manipulated to improve milk yield and quality. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1522-1533, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27017   open full text
• Role of STAT signaling and autocrine action of chemokines during H2O 2 induced HTR‐8/SVneo trophoblastic cells invasion.
  Priyanka Banerjee, Ankita Malik, Sudha Saryu Malhotra, Satish Kumar Gupta.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- H2O2 (a reactive oxygen species) at 25 μM concentration increases human HTR‐8/SVneo cells invasion. The underlying mechanism of H2O2‐mediated increase in HTR‐8/SVneo cells invasion depends on increased MMP‐9/TIMP‐1 ratio, and secretion of IL‐8 and MIP‐1β via activation of STAT signaling. During pregnancy, regulated generation of reactive oxygen species (ROS) is important for activation of signaling pathways and placentation. In the current study, the effect of H2O2 on invasion of HTR‐8/SVneo cells, a human extravillous trophoblast cell line, is investigated. Treatment of HTR‐8/SVneo cells for 24 hr with H 2O2 (25 µM) leads to a significant increase in invasion without affecting cell proliferation, viability, and apoptosis. Concomitantly, a significant increase in the matrix metalloproteinase‐9 (MMP‐9)/tissue inhibitor of metalloproteinases‐1 (TIMP‐1) ratio is observed. Further, significant increase in phosphorylation of signal transducer and activator of transcription 1 (STAT‐1) and STAT‐3 (both at ser727 residue) is observed on treating HTR‐8/SVneo cells with 25 µM of H2O2 accompanied by an increase in the secretion of interleukin‐8 (IL‐8) and macrophage inflammatory protein‐1β (MIP‐1β). A significant decrease in H2O2‐mediated invasion of HTR‐8/SVneo cells and reduced expression of IL‐8 and MIP‐1β accompanied by decrease in MMP‐9/TIMP‐1 ratio are observed on inhibiting STAT‐1 and STAT‐3 by small interfering RNA (siRNA). Inhibition of STAT‐1 activity by fludarabine and STAT‐3 activity by Stattic also leads to a decrease in H2O2‐mediated increase in HTR‐8/SVneo cell invasion. Inhibition of IL‐8 and MIP‐1β by siRNA also leads to a significant decrease in both basal and H2O2‐mediated invasion. Interestingly, inhibition of MIP‐1β by siRNA leads to a significant reduction in H2O2‐mediated increase in IL‐8. However, no significant effect of IL‐8 silencing on H2O2‐mediated MIP‐1β expression was observed. From the above results, it can be concluded that H2O2 activates STAT signaling, MIP‐1β & IL‐8 secretion and increases MMP‐9/TIMP‐1 ratio leading to an increased invasion of HTR‐8/SVneo cells without affecting their viability. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1380-1397, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26934   open full text
• Hypoxic conditioned medium derived from bone marrow mesenchymal stromal cells protects against ischemic stroke in rats.
  Run‐Hao Jiang, Chen‐Jiang Wu, Xiao‐Quan Xu, Shan‐Shan Lu, Qing‐Quan Zu, Lin‐Bo Zhao, Jun Wang, Sheng Liu, Hai‐Bin Shi.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Conditioned medium constitutes a therapeutic effect on stroke. Paracrine actions of bone marrow mesenchymal stromal cell are enhanced by hypoxic preconditioning. Apoptosis and neovascularization are involved in this beneficial effect. In recent years, studies have shown that the secretome of bone marrow mesenchymal stromal cells (BMSCs) contains many growth factors, cytokines, and antioxidants, which may provide novel approaches to treat ischemic diseases. Furthermore, the secretome may be modulated by hypoxic preconditioning. We hypothesized that conditioned medium (CM) derived from BMSCs plays a crucial role in reducing tissue damage and improving neurological recovery after ischemic stroke and that hypoxic preconditioning of BMSCs robustly improves these activities. Rats were subjected to ischemic stroke by middle cerebral artery occlusion and then intravenously administered hypoxic CM, normoxic CM, or Dulbecco modified Eagle medium (DMEM, control). Cytokine antibody arrays and label‐free quantitative proteomics analysis were used to compare the differences between hypoxic CM and normoxic CM. Injection of normoxic CM significantly reduced the infarct area and improved neurological recovery after stroke compared with administering DMEM. These outcomes may be associated with the attenuation of apoptosis and promotion of angiogenesis. Hypoxic preconditioning significantly enhanced these therapeutic effects. Fourteen proteins were significantly increased in hypoxic CM compared with normoxic CM as measured by cytokine arrays. The label‐free quantitative proteomics analysis revealed 163 proteins that were differentially expressed between the two groups, including 107 upregulated proteins and 56 downregulated proteins. Collectively, our results demonstrate that hypoxic CM protected brain tissue from ischemic injury and promoted functional recovery after stroke in rats and that hypoxic CM may be the basis of a potential therapy for stroke patients. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1354-1368, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26931   open full text
• Src mediates TGF‐β‐induced intraocular pressure elevation in glaucoma.
  Teruhisa Tsukamoto, Kentaro Kajiwara, Shigeyuki Nada, Masato Okada.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Src kinase is involved in TGF‐β‐induced elevation of intraocular pressure in glaucoma. Src regulates TGF‐β2‐induced changes in the contractile and adhesive characteristics of trabecular meshwork cells, and ECM deposition. Glaucoma, a progressive and irreversible optic neuropathy, is one of the leading causes of vision impairment worldwide. Elevation of intraocular pressure (IOP) due to transforming growth factor‐β (TGF‐β)‐induced dysfunction of the trabecular meshwork is a risk factor for glaucoma, but the underlying molecular mechanisms remain elusive. Here, we show that Src kinase is involved in TGF‐β‐induced IOP elevation. We observed that dasatinib, a potent Src inhibitor, suppressed TGF‐β2‐induced IOP in rat eyes. Mechanistic analyses in human trabecular meshwork cells showed that TGF‐β2 activated Src signaling and concomitantly increased cytoskeletal remodeling, cell adhesion, and extracellular matrix (ECM) accumulation. Src was activated via TGF‐β2‐induced upregulation of the Src scaffolding protein CasL, which mediates the assembly of focal adhesions, cytoskeletal remodeling, and ECM deposition. Activation of Src suppressed the expression of tissue plasminogen activator, thereby attenuating ECM degradation. Furthermore, the Src inhibitor ameliorated TGF‐β2‐induced changes in the contractile and adhesive characteristics of trabecular meshwork cells, and ECM deposition. These findings underscore the crucial role of Src activity in TGF‐β‐induced IOP elevation and identify Src signaling as a potential therapeutic target in glaucoma. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1730-1744, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27044   open full text
• Deletion of Axin1 in condylar chondrocytes leads to osteoarthritis‐like phenotype in temporomandibular joint via activation of β‐catenin and FGF signaling.
  Yachuan Zhou, Bing Shu, Rong Xie, Jian Huang, Liwei Zheng, Xuedong Zhou, Guozhi Xiao, Lan Zhao, Di Chen.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- We have generated Axin1 conditional knockout (KO) mice by targeting aggrecan‐expressing condylar cartilage cells in the temporomandibular joint (TMJ). The Axin1 conditional KO mice showed TMJ osteoarthritis (OA)‐like phenotype, probably through activation of β‐catenin and fibroblast growth factor (FGF) signaling. Osteoarthritis (OA) in the temporomandibular joint (TMJ) is a degenerative disease in the adult, which is characterized by the pathological degeneration of condylar cartilage. Axin1 plays a critical role in the regulation of cartilage development and homeostasis. To determine the role of Axin1 in TMJ tissue at the adult stage, we generated Axin1Agc1ER mice, in which Axin1 was deleted in aggrecan‐expressing chondrocytes at 2 months of age. Histology, histomorphometry, and immunostaining analyses were performed using TMJ tissues harvested from 4‐ and 6‐month‐old mice after tamoxifen administration. Total RNA isolated from TMJ cartilage of 6‐month‐old mice was used for gene expression analysis. Progressive OA‐like degeneration was observed in condylar cartilage in Axin1 knockout (KO) mice with loss of surface continuity and the formation of vertical fissures. In addition, reduced alcian blue staining in condylar cartilage was also found in Axin1 KO mice. Immunostaining and reverse transcription quantitative polymerase chain reaction (qRT‐PCR) assays revealed disturbed homeostasis in condylar cartilage with increased expressions of MMP13 and Adamts5 and decreased lubricin expression in Axin1‐deficient chondrocytes. Less proliferative cells with increased hypertrophic and apoptotic activities were presented in the condylar cartilage of Axin1Agc1ER KO mice. As a scaffolding protein, the deletion of Axin1 stimulated not only the β‐catenin but also the fibroblast growth factor (FGF) signaling via extracellular signal‐regulated protein kinases 1 and 2 (ERK1/2) activation. The qRT‐PCR results showed an increased expression of Fgfr1 in Axin1 KO cartilage. Overall, the deletion of Axin1 in condylar chondrocytes altered the β‐catenin and FGF/ERK1/2 signaling pathways, thus cooperatively contribute to the cartilage degeneration. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1720-1729, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27043   open full text
• Effects of carnitine palmitoyltransferases on cancer cellular senescence.
  Lihuan Guan, Yixin Chen, Yongtao Wang, Huizhen Zhang, Shicheng Fan, Yue Gao, Tingying Jiao, Kaili Fu, Jiahong Sun, Aiming Yu, Min Huang, Huichang Bi.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- This study revealed that carnitine palmitoyltransferase 1C (CPT1C) knockdown exhibited the strongest impact on cell growth arrest and senescence, lipid change and accumulation, and mitochondrial dysfunction among all members of the CPT family. In addition, only knocking down CPT1C could downregulate the protein expression of c‐Myc and cyclin D1 and upregulate the cell cycle inhibitor p27 significantly, which contributes to the potential molecular mechanism of CPT1C knockdown‐induced cellular senescence. The carnitine palmitoyltransferase (CPT) family is essential for fatty acid oxidation. Recently, we found that CPT1C, one of the CPT1 isoforms, plays a vital role in cancer cellular senescence. However, it is unclear whether other isoforms (CPT1A, CPT1B, and CPT2) have the same effect on cellular senescence. This study illustrates the different effects of CPT knockdown on PANC‐1 cell proliferation and senescence and MDA‐MB‐231 cell proliferation and senescence, as demonstrated by cell cycle kinetics, Bromodeoxyuridine incorporation, senescence‐associated β‐galactosidase activity, colony formation, and messenger RNA (mRNA) expression of key senescence‐associated secretory phenotype factors. CPT1C exhibits the most substantial effect on cell senescence. Lipidomics analysis was performed to further reveal that the knockdown of CPTs changed the contents of lipids involved in mitochondrial function, and lipid accumulation was induced. Moreover, the different effects of the isoform deficiencies on mitochondrial function were measured and compared by the level of radical oxygen species, mitochondrial transmembrane potential, and the respiratory capacity, and the expression of the genes involved in mitochondrial function were determined at the mRNA level. In summary, CPT1C exerts the most significant effect on mitochondrial dysfunction‐associated tumor cellular senescence among the members of the CPT family, which further supports the crucial role of CPT1C in cellular senescence and suggests that inhibition of CPT1C may represent as a new strategy for cancer treatment through the induction of tumor senescence. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1707-1719, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27042   open full text
• Implication of basal lamina dependency in survival of Nrf2‐null muscle stem cells via an antioxidative‐independent mechanism.
  Yusei Takemoto, Shoya Inaba, Lidan Zhang, Kazutake Tsujikawa, Akiyoshi Uezumi, So‐ichiro Fukada.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Muscle regeneration ability of Nrf2‐null mice is normal. However, in a basal lamina–disrupted model, Nrf2‐null mice exhibited remarkable regeneration defects without increased levels of reactive oxidative species in muscle stem cells Nuclear factor erythroid 2–related factor 2 (Nrf2) is a master regulator for the induction of antioxidative genes and plays roles in diverse cellular functions. The roles of Nrf2 in muscle regeneration have been investigated, and both important and unimportant roles of Nrf2 for muscle regeneration have been reported. Here, using aged Nrf2‐null and Nrf2–dystrophic double‐null mice, we showed nonsignificant phenotypes in the muscle regeneration ability of Nrf2‐null mice. In contrast with these results, strikingly, almost all Nrf2‐null muscle stem cells (MuSCs) isolated by fluorescence‐activated cell sorting died in vitro of apoptosis and were not rescued by antioxidative reagents. Although their proliferation was still impaired, the Nrf2‐null MuSCs attached to myofibers activated and divided normally, at least in the first round. To elucidate these discrepancies of MuSCs behaviors, we focused on the basal lamina, because both in vivo and single myofiber culture allow MuSCs within the basal lamina to become activated. In a basal lamina–disrupted model, Nrf2‐null mice exhibited remarkable regeneration defects without increased levels of reactive oxidative species in MuSCs, suggesting that the existence of the basal lamina affects the survival of Nrf2‐null MuSCs. Taken together, these results suggest that the basal lamina compensates for the loss of Nrf2, independent of the antioxidative roles of Nrf2. In addition, experimental conditions might explain the discrepant results of Nrf2‐null regenerative ability. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1689-1698, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27040   open full text
• Protective effects of ex‐527 on cerebral ischemia–reperfusion injury through necroptosis signaling pathway attenuation.
  Sara Nikseresht, Fariba Khodagholi, Abolhassan Ahmadiani.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- The selective and potent inhibitor of Sirt1, ex‐527, reduces the infarction volume of ischemic brains and improves the survival rate. Treatment with ex‐527 effectively abolishes the elevation of the critical regulators of necroptosis. Ex‐527 relieves ischemia‐induced perturbation of necroptosis‐associated metabolic enzyme activity downstream. Necroptosis, a novel type of programmed cell death, is involved in ischemia–reperfusion‐induced brain injury. Sirtuin 1 (Sirt1), as a well‐known member of histone deacetylase class III, plays pivotal roles in inflammation, metabolism, and neuron loss in cerebral ischemia. We explored the relationship between Sirt1 and the necroptosis signaling pathway and its downstream events by administration of ex‐527, as a selective and potent inhibitor of Sirt1, and necrostatin‐1 (nec‐1), as a necroptosis inhibitor, in an animal model of focal cerebral ischemia. Our data showed different patterns of sirt1 and necroptosis critical regulators, including receptor‐interacting protein kinase 3 and mixed lineage kinase domain–like protein gene expressions in the prefrontal cortex and the hippocampus after ischemia–reperfusion. We found that ex‐527 microinjection reduces the infarction volume of ischemic brains and improves the survival rate, but not stroke‐associated neurological deficits. Additionally, treatment with ex‐527 effectively abolished the elevation of the critical regulators of necroptosis, whereas necroptosis inhibition through nec‐1 microinjection did not influence Sirt1 expression levels. Our data also demonstrated that the ex‐527 relieves ischemia‐induced perturbation of necroptosis‐associated metabolic enzymes activity in downstream. This study provides a new approach to the possible neuroprotective potential of ex‐527 orchestrated by necroptosis pathway inhibition to alleviate ischemia–reperfusion brain injury. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1816-1826, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27055   open full text
• Exercise protects against diabetic cardiomyopathy by the inhibition of the endoplasmic reticulum stress pathway in rats.
  Wang Chengji, Fan Xianjin.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Exercise training improves diabetic cardiomyopathy and heart function by decreasing the cardiac myocyte ER stress and subsequent myocardial apoptosis.This is an important finding because ER stress is one of the underlying mechanisms of diabetic complications. Thus, exercise training may significantly contribute to complication prevention in diabetes, which shows an intensity effect. Our findings reveal a novel mechanism of the effect of exercise in the management of diabetic cardiomyopathy. To explore the protective effect of exercise training on the injury of myocardium tissues induced by streptozotocin (STZ) in diabetic rats and the relationship with endoplasmic reticulum stress (ERS), the male sprague‐dawley (SD) rats were fed with high‐fat and high‐sugar diet for 4 weeks, followed by intraperitoneal injection of STZ, 40 mg/kg, to establish a diabetes model, and then 10 rats were randomly selected as diabetes mellitus (DM) controls and 20 eligible diabetic rats were randomized into two groups: low‐intensity exercise training (n = 10) and high‐intensity exercise training (n = 10). After 12 weeks of exercise training, rats were killed and serum samples were used to determine cardiac troponin‐I (cTn‐I). Myocardial tissues were sampled for morphological analysis to detect myocardial cell apoptosis, and to analyze protein expression of glucose‐regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), and caspase‐12. Different intensities (low and high) significantly reduced serum cTn‐I levels compared with the DCM group (p < 0.01), and significantly reduced the percentage of apoptotic myocardial cells and improved the parameters of cardiac function. Hematoxylin and eosin and Masson staining indicated that exercise training could attenuate myocardial apoptosis. Additionally, exercise training significantly reduced GRP78, CHOP, and cleaved caspase‐12 protein expression in an intensity‐dependent manner. These findings suggest that exercise appeared to ameliorate diabetic cardiomyopathy by inhibiting endoplasmic reticulum stress‐induced apoptosis in diabetic rats. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1682-1688, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27038   open full text
• Gastrin‐releasing peptide receptor gene silencing inhibits the development of the epithelial–mesenchymal transition and formation of a calcium oxalate crystal in renal tubular epithelial cells in mice with kidney stones via the PI3K/Akt signaling pathway.
  Xin‐Fang Wang, Bei‐Hao Zhang, Xiao‐Qing Lu, Rui‐Qiang Wang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Gastrin‐releasing peptide receptor gene silencing suppresses the development of the epithelial–mesenchymal transition (EMT) and formation of a calcium oxalate crystal in renal tubular epithelial cells (TECs) of kidney stones through the inactivation of the PI3K/Akt signaling pathway. Between 1% and 15% of people are globally affected by kidney stones, and this disease has become more common since the 1970s. Therefore, this study aims to investigate the effects of gastrin‐releasing peptide receptor (GRPR) gene silencing via the PI3K/Akt signaling pathway on the development of the epithelial–mesenchymal transition (EMT) and formation of a calcium oxalate crystal in renal tubular epithelial cells (TECs) of kidney stones. A total of 70 clean and healthy C57BL/6J mice were assigned into the normal ( n = 10) and kidney stones groups ( n = 60). The underlying regulatory mechanisms of GRPR were analyzed in concert with the treatment of shGRPR‐1, LY294002, and shGRPR‐1 + LY294002 in TECs isolated from mice with kidney stones. A series of experiments were conducted for the measurement of urinary oxalate and urinary calcium, the renal calcium salt deposition, the positive rate of GRPR, the expressions of renal TECs related genes and calcium oxalate regulation related genes, and the growth of calcium crystals induced by cells. After treatment of shGRPR‐1 and shGRPR‐1 + LY294002, levels of urinary oxalate and urinary calcium in the serum, as well as positive rate of GRPR, became relatively low, levels of E‐cadherin enhanced, whereas levels of Akt, PI3K, GRPR, extents of PI3K and Akt phosphorylation, α‐SMA, Vimentin and FSP‐1, OPN, MCP‐1, and CD44 decreased and a number of crystals reduced. Taken together, we conclude that GRPR gene silencing suppresses the development of the EMT and formation of the calcium oxalate crystal in renal TECs of kidney stones through the inactivation of the PI3K/Akt signaling pathway. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1567-1577, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27023   open full text
• Combination therapy with KRAS siRNA and EGFR inhibitor AZD8931 suppresses lung cancer cell growth in vitro.
  Habib Zarredar, Shadi Pashapour, Khalil Ansarin, Majid Khalili, Roghayyeh Baghban, Safar Farajnia.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this study, we try to understand that if we knock down some important genes like the Kirsten rat sarcoma oncogene (KRAS) and the epidermal growth factor receptor (EGFR) family in lung cancer cell lines, we could control proliferation and development of these cancer cells. Lung cancer is a leading cause of cancer‐related deaths worldwide, with less than a 5‐year survival rate for both men and women. Epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma oncogene (KRAS) signaling pathways play a critical role in the proliferation and progression of various cancers, including lung cancer. Genetic studies have shown that amplification, over‐expression, or mutation of EGFR is an early and major molecular event in many human tumors. KRAS mutation is a negative factor in various cancer, including non‐small‐cell lung cancer, and complicates therapeutic approaches with adjuvant chemotherapy and anti‐EGFR directed therapies. This article is dedicated to evaluating the synergistic effect of a novel EGFR inhibitor AZD8931 and KRAS small interfering RNA (siRNA) on the proliferation and apoptosis of lung adenocarcinoma cancer cells. A549 lung cancer cells were treated with KRAS siRNA and the EGFR inhibitor alone or in combination. The cytotoxic effects of KRAS siRNA and te EGFR inhibitor were determined usingMTT assay, and induction of apoptosis was determined by FACS analysis. Suppression of KRAS, Her‐2, and EGFR expression by treatments was measured by qRT‐PCR and western blotting. KRAS siRNA and the EGFR inhibitor significantly reduced the proliferation of A549 cells as well as KRAS and EGFR mRNA levels 24 hr after treatment. The results also indicated that the silencing of KRAS and EGFR has synergistic effects on the induction of apoptosis on the A549 cells. These results indicated that KRAS and EGFR might play important roles in the progression of lung cancer and could be potential therapeutic targets for treatment of lung cancer. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1560-1566, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27021   open full text
• Inhibition of JNK and p38 MAPK‐mediated inflammation and apoptosis by ivabradine improves cardiac function in streptozotocin‐induced diabetic cardiomyopathy.
  Guangfeng Zuo, Xiaomin Ren, Xuesong Qian, Peng Ye, Jie Luo, Xiaofei Gao, Junjie Zhang, Shaoliang Chen.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 (a) The JNK and p38 MAPK protein levels in heart tissues after intramyocardial injection of lentiviruses carrying shRNA targeting JNK (LV‐JNK shRNA) and p38 MAPK (LV‐p38 MAPK shRNA). (b,c) Proinflammatory cytokine mRNA and protein expression were significantly decreased in diabetic mice treated with LV‐JNK shRNA and LV‐p38 MAPK shRNA. Column figure shows the difference in mRNA and protein expression. (d) TUNEL staining of heart tissues in each group. Inflammation plays a critical role in the development of diabetic cardiomyopathy (DCM), which has been identified as a major predisposing factor for heart failure in diabetic patients. Previous studies indicated that ivabradine (a specific agent for heart rate [HR] reduction) has anti‐inflammatory properties, but its role in DCM remains unknown. This study investigated whether ivabradine exerts a therapeutic effect in DCM. C57BL/6J mice were injected intraperitoneally with streptozotocin (STZ) to induce diabetes; then administered with ivabradine or saline (control). After 12 weeks, the surviving mice were analyzed to determine the cardioprotective effect of ivabradine against DCM. Although treatment with ivabradine did not affect blood glucose levels, it attenuated tumor necrosis factor‐α, interleukin‐1β, and interleukin‐6 messenger RNA (mRNA) expression, inhibited c‐Jun N‐terminal kinase (JNK) and p38 mitogen‐activated protein kinase (p38 MAPK) activation, reduced histological abnormalities, myocardial apoptosis and collagen deposition, and improved cardiac function in the diabetic mice. Interestingly, the anti‐inflammatory and antiapoptotic properties of ivabradine, but not its inhibitory effect on JNK and p38 MAPK, were observed in high‐glucose‐cultured neonatal rat ventricular cardiomyocytes. Attenuating inflammation and apoptosis via intramyocardial injection of lentiviruses carrying short hairpin RNA targeting JNK and p38 MAPK validated that the anti‐inflammatory and antiapoptotic effects of ivabradine were partly attributed to JNK and p38 MAPK inactivation in diabetic mice. In summary, these data indicate that ivabradine‐mediated improvement of cardiac function in STZ‐induced diabetic mice may be partly attributed to inhibition of JNK/p38 MAPK‐mediated inflammation and apoptosis, which is dependent on the reduction in HR. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1925-1936, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27070   open full text
• Identification of key gene modules and pathways of human glioma through coexpression network.
  Tianming Shi, Jianchun Chen, Jing Li, Bing‐Yin Yang, Qiao‐Lin Zhang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Glioma causes great harm to people worldwide. Systemic coexpression analysis of this disease could be beneficial for the identification and development of new prognostic and predictive markers in the clinical management of glioma. In this study, we extracted data sets from GEO (Gene Expression Omnibus) data set by using “glioma” as the keyword. Then, a coexpression module was constructed with the help of WGCNA (Weighted Gene Coexpression Network Analysis) software. Besides, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the genes in these modules. As a result, the critical modules and target genes were identified. Eight coexpression modules were constructed using the 4,000 genes with a high expression value of the total 141 glioma samples. The result of the analysis of the interaction among these modules showed that there was a high scale independence degree among them. The GO and KEGG enrichment analyses showed that there was a significant difference in the enriched terms and degree among these eight modules, and module 5 was identified as the most important module. Besides, the pathways it was enriched in, hsa04510: Focal adhesion and hsa04610: Complement and coagulation cascades, were determined as the most important pathways. In summary, module 5 and the pathways it was enriched in, hsa04510: Focal adhesion and has 04610: Complement and coagulation cascades, have the potential to serve as biomarkers for glioma patients. Glioma causes great harm to people worldwide. Systemic coexpression analysis of this disease could be beneficial for the identification and development of new prognostic and predictive markers in the clinical management of glioma. In this study, we extracted data sets from the Gene Expression Omnibus data set by using “glioma” as the keyword. Then, a coexpression module was constructed with the help of Weighted Gene Coexpression Network Analysis software. Besides, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the genes in these modules. As a result, the critical modules and target genes were identified. Eight coexpression modules were constructed using the 4,000 genes with a high expression value of the total 141 glioma samples. The result of the analysis of the interaction among these modules showed that there was a high scale independence degree among them. The GO and KEGG enrichment analyses showed that there was a significant difference in the enriched terms and degree among these eight modules, and module 5 was identified as the most important module. Besides, the pathways it was enriched in, hsa04510: Focal adhesion and hsa04610: Complement and coagulation cascades, were determined as the most important pathways. In summary, module 5 and the pathways it was enriched in, hsa04510: Focal adhesion and has 04610: Complement and coagulation cascades, have the potential to serve as biomarkers for patients with glioma. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1862-1870, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27059   open full text
• The effect of high‐fat diet and inhibition of ceramide production on insulin action in liver.
  Piotr Zabielski, Jarosław Daniluk, Hady Razak Hady, Adam R. Markowski, Monika Imierska, Jan Górski, Agnieszka U. Blachnio‐Zabielska.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Hepatic insulin resistance induced by high‐fat diet resulted from an increase in synthesis rate of ceramide and diacylglycerol (DAG). Myriocin treatment improved insulin sensitivity, which was accompanied by a decrease in the synthesis rate and content of ceramide and DAG. Liver, as one of the most important organs involved in lipids and glucose metabolism, is perceived as a key tissue for pharmacotherapy of insulin resistance (IRes) and type 2 diabetes. Ceramides (Cer) are biologically active lipids, which accumulation is associated with the induction of muscle IRes. We sought to determine the role of intrahepatic bioactive lipids production on insulin action in liver of insulin‐resistant rats and after myriocin administration. The experiments were conducted on male Wistar rats divided into three groups: Control, fed high‐fat diet (HFD), and fed HFD and treated with myriocin (HFD/Myr). Before sacrifice, the animals were infused with a [U‐13C]palmitate to calculate lipid synthesis rate by means of tracer incorporation technique in particular lipid groups. Liver Cer, diacylglycerols (DAG), acyl‐carnitine concentration, and isotopic enrichment were analyzed by LC/MS/MS. Proteins involved in lipid metabolism and insulin pathway were analyzed by western blot analysis. An OGTT and ITT was also performed. HFD‐induced IRes and increased both the synthesis rate and the content of DAG and Cer, which was accompanied by inhibition of an insulin pathway. Interestingly, myriocin treatment reduced synthesis rate not only of Cer but also DAG and improved insulin sensitivity. We conclude that the insulin‐sensitizing action of myriocin in the liver is a result of the lack of inhibitory effect of lipids on the insulin pathway, due to the reduction of their synthesis rate. This is the first study showing how the synthesis rate of individual lipid groups in liver changes after myriocin administration. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1851-1861, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27058   open full text
• MicroRNA‐590‐5p functions as a tumor suppressor in breast cancer conferring inhibitory effects on cell migration, invasion, and epithelial–mesenchymal transition by downregulating the Wnt–β‐catenin signaling pathway.
  Jin Gao, Shao‐Rong Yu, Yuan Yuan, Li‐Li Zhang, Jian‐Wei Lu, Ji‐Feng Feng, Sai‐Nan Hu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- This study provides evidence suggesting that the transfection of overexpressed miR‐590‐5p can act to alleviate the effects of breast cancer demonstrating an ability to inhibit the processes of cell proliferation, migration, and invasion as well as epithelial–mesenchymal transition by suppressing the expression of paired‐like homeodomain transcription factor 2 and activation of the Wnt–β‐catenin pathway. Abstract Breast cancer remains one of the foremost primary causes of female morbidity and mortality worldwide. During the current study, the effect of miR‐590‐5p and paired‐like homeodomain transcription factor 2 (PITX2) on proliferation, invasion, migration, and epithelial–mesenchymal transition (EMT) of human breast cancer via the Wnt–β‐catenin signaling pathway was investigated. Breast cancer–related genes and related signaling pathways were obtained from KEGG database. The PITX2 regulatory microRNA was predicted. To define the contributory role by which miR‐590‐5p influences the progression of breast cancer, the interaction between miR‐590‐5p and PITX2 was explored; the proliferation, invasion, and migration abilities as well as the tumor growth and metastasis in nude mice were detected following the overexpression or silencing of miR‐590‐5p. PITX2 was determined to share a correlation with breast cancer and miR‐590‐5p was selected for further analysis. PITX2, Wnt‐1, β‐catenin, N‐cadherin, and vimentin all displayed higher levels, while miR‐590‐5p and E‐cadherin expression were lower among breast cancer tissues than in the adjacent normal tissue. After overexpression of miR‐590‐5p or si‐PITX2, the expression of E‐cadherin was markedly increased, decreases in the expression of Wnt‐1, β‐catenin, N‐cadherin, and vimentin, as well as inhibited cell proliferation, invasion, migration, metastasis, and EMT were observed. This study provides evidence suggesting that the transfection of overexpressed miR‐590‐5p can act to alleviate the effects of breast cancer demonstrating an ability to inhibit the processes of cell proliferation, migration, and invasion as well as EMT by suppressing the expression of PITX2 and activation of the Wnt–β‐catenin pathway. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1827-1841, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27056   open full text
• Licochalcone D directly targets JAK2 to induced apoptosis in human oral squamous cell carcinoma.
  Ji‐Hye Seo, Hyun Woo Choi, Ha‐Na Oh, Mee‐Hyun Lee, Eunae Kim, Goo Yoon, Seung‐Sik Cho, Seon‐Min Park, Young Sik Cho, Jung‐Il Chae, Jung‐Hyun Shim.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Our results demonstrated that LCD inhibited the kinase activity of JAK2, soft agar colony formation, and the proliferation of HN22 and HSC4 cells. LCD also induced mitochondrial apoptotic events such as altered mitochondrial membrane potential and reactive oxygen species production and, LCD increased expression of apoptosis‐associated proteins in oral squamous cell carcinoma cells. These results indicate that the anticancer effect of LCD is due to the direct targeting of JAK2 kinase. Licochalcone (LC) families have been reported to have a wide range of biological function such as antioxidant, antibacterial, antiviral, and anticancer effects. Although various beneficial effects of LCD were revealed, its anticancer effect in human oral squamous cancer has not been identified. To examine the signaling pathway of LCD’s anticancer effect, we determined whether LCD has physical interaction with Janus kinase (JAK2)/signal transducer and activator of transcription‐3 (STAT3) signaling, which is critical in promoting cancer cell survival and proliferation. Our results demonstrated that LCD inhibited the kinase activity of JAK2, soft agar colony formation, and the proliferation of HN22 and HSC4 cells. LCD also induced mitochondrial apoptotic events such as altered mitochondrial membrane potential and reactive oxygen species production. LCD increased the expression of apoptosis‐associated proteins in oral squamous cell carcinoma (OSCC) cells. Finally, the xenograft study showed that LCD significantly inhibited HN22 tumor growth. Immunohistochemical data supported that LCD suppressed p‐JAK2 and p‐STAT3 expression and induced cleaved‐caspase‐3 expression. These results indicate that the anticancer effect of LCD is due to the direct targeting of JAK2 kinase. Therefore, LCD can be used for therapeutic application against OSCC. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1780-1793, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27050   open full text
• LncRNA PVT1 regulates VEGFC through inhibiting miR‐128 in bladder cancer cells.
  Cui Yu, Liu Longfei, Wang Long, Zeng Feng, Jinbo Chen, Li Chao, Liu Peihua, Zu Xiongbing, Chen Hequn.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- PVT1 might play a critical role in bladder cancer tumorigenesis via miR‐218 and VEGFC. PVT1 could be a new biomarker for bladder cancer diagnosis and therapy. Long noncoding RNA PVT1 is considered to be an oncogene in multiple cancers. Our previous studies indicated that PVT1 levels were higher in bladder cancer tissue and correlated with clinical progression and poor prognosis in bladder cancer patients. A bioinformatics analysis showed that PVT1 may regulate VEGFC expression through miR‐128 as a competing endogenous RNA (ceRNA). In this study, we demonstrated that PVT1 expression levels affect the proliferation and migration ability of bladder cancer cells. Moreover, PVT1 knockdown significantly decreased the proliferation capacity of bladder cancer cells in nude mice. Luciferase assays and RNA‐binding protein immunoprecipitation were performed to investigate the potential mechanism of ceRNAs in the regulation of PVT1 and VEGFC. The results showed that the increased number of PVT1 transcripts interacted directly with miR‐128 to decrease miR‐128 binding to the VEGFC 3′‐untranslated region. This effect suppressed VEGFC mRNA degradation by miR‐128. In conclusion, these results indicated that PVT1 might play a critical role in bladder cancer tumorigenesis via miR‐218 and VEGFC. Therefore, PVT1 could be a new biomarker for bladder cancer diagnosis and therapy. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1346-1353, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26929   open full text
• Long non‐coding RNA LINC00152 promotes cell growth and invasion of papillary thyroid carcinoma by regulating the miR‐497/BDNF axis.
  Zhihui Sun, Xun Guo, Mingcui Zang, Peisong Wang, Shuai Xue, Guang Chen.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 Long intergenic non‐coding RNA 152 (LINC00152) was reported to be tightly linked to tumorigenesis and progression in multiple cancers. However, its biological role and modulatory mechanism in papillary thyroid carcinoma (PTC) has not been elucidated. In this study, we determined the expression levels of LINC00152 in PTC tissues and cell lines by quantitative real time polymerase chain reaction (qRT‐PCR). Cell proliferation, colony formation, migration, and invasion were measured by a Cell Counting Kit‐8 assay, colony formation analysis, wound healing, and transwell invasion assay, respectively. A luciferase reporter assay and qRT‐PCR were used to determine whether LINC00152 interacts with miR‐497 directly. We established a xenograft mouse model to examine the underlying molecular mechanism and effect of LINC00152 on tumor growth in vivo. We found that LINC00152 expression was significantly increased in PTC tissues and derived cell lines. LINC00152 knockdown significantly inhibited proliferation, colony formation, migration, and invasion in vitro, and impaired tumor growth in vivo. We revealed that LINC00152 functioned as a competing endogenous RNA to the miR‐497 sponge, downregulating its downstream target brain‐derived neurotrophic factor (BDNF), which is an oncogene in thyroid cancer. These findings suggest that LINC00152 is responsible for PTC cell proliferation and invasion and exerts its function by regulating the miR‐497/BDNF axis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1336-1345, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26928   open full text
• Estrogen‐related receptor γ negatively regulates osteoclastogenesis and protects against inflammatory bone loss.
  Hyun‐Ju Kim, Bo Kyung Kim, Boram Ohk, Hye‐Jin Yoon, Woo Youl Kang, Seungil Cho, Sook Jin Seong, Hae Won Lee, Young‐Ran Yoon.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Our findings identify orphan nuclear receptor ERRγ as a novel negative regulator of osteoclastogenesis and further reveals its protective role on inflammatory bone loss in vivo. Estrogen‐related receptor γ (ERRγ) is an orphan nuclear receptor that plays an important role in various metabolic processes under physiological and pathophysiological conditions. Here, we report that ERRγ functions as a negative regulator in receptor activator of nuclear factor κΒ ligand (RANKL)–induced osteoclast differentiation. We observed that ERRγ was strongly expressed in osteoclast precursors, bone marrow–derived macrophages (BMMs) while its expression was significantly reduced by RANKL during osteoclastogenesis. Overexpression of ERRγ in BMMs suppressed the formation of multinucleated osteoclasts and attenuated the induction of c‐Fos and nuclear factor of activated T cells c1, which are critical modulators in osteoclastogenesis. Similarly, the treatment of ERRγ agonists, N‐(4‐(diethylaminobenzylidenyl)‐N'‐(4‐hydroxybenzoyl)‐hydrazine (DY131) or GSK4716, also inhibited osteoclast generation and the expression of these key modulators. On the other hand, shRNA‐mediated knockdown of ERRγ accelerated the formation of bone‐resorbing cells and the expression of osteoclastogenic markers. Forced expression of ERRγ blocked RANKL‐stimulated phosphorylation of the nuclear factor κB (NF‐κB) inhibitor IκBα and suppressed NF‐κB transcriptional activity induced by RANKL or the NF‐κB subunit p65. Furthermore, by employing a pharmacological approach, we showed that the ERRγ agonist DY131 protected against inflammatory bone loss induced by lipopolysaccharide in vivo. Together, our findings reveal that ERRγ is a pivotal regulator in RANKL‐mediated osteoclastogenesis and suggest that ERRγ may have potential as a therapeutic target for pathological bone loss. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1659-1670, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27035   open full text
• Antioxidant effect of human placenta hydrolysate against oxidative stress on muscle atrophy.
  Dong‐ho Bak, Jungtae Na, Song I Im, Chang Taek Oh, Jeom‐Yong Kim, Sun‐Kyu Park, Hae Jung Han, Joon Seok, Sun Young Choi, Eun Jung Ko, Seog‐Kyun Mun, Suk‐Won Ahn, Beom Joon Kim.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Human placental hydrolysate required for the regulation of oxidative stress through the control of Mstn expression by nuclear factor‐κB, mitochondrial quality control, and autophagy process, leading to improved muscle atrophy. Sarcopenia, which refers to the muscle loss that accompanies aging, is a complex neuromuscular disorder with a clinically high prevalence and mortality. Despite many efforts to protect against muscle weakness and muscle atrophy, the incidence of sarcopenia and its related permanent disabilities continue to increase. In this study, we found that treatment with human placental hydrolysate (hPH) significantly increased the viability (approximately 15%) of H2O2‐stimulated C2C12 cells. Additionally, while H2O2‐stimulated cells showed irregular morphology, hPH treatment restored their morphology to that of cells cultured under normal conditions. We further showed that hPH treatment effectively inhibited H2O2‐induced cell death. Reactive oxygen species (ROS) generation and Mstn expression induced by oxidative stress are closely associated with muscular dysfunction followed by atrophy. Exposure of C2C12 cells to H2O2 induced abundant production of intracellular ROS, mitochondrial superoxide, and mitochondrial dysfunction as well as myostatin expression via nuclear factor‐κB (NF‐κB) signaling; these effects were attenuated by hPH. Additionally, hPH decreased mitochondria fission–related gene expression (Drp1 and BNIP3) and increased mitochondria biogenesis via the Sirt1/AMPK/PGC‐1α pathway and autophagy regulation. In vivo studies revealed that hPH‐mediated prevention of atrophy was achieved predominantly through regulation of myostatin and PGC‐1α expression and autophagy. Taken together, our findings indicate that hPH is potentially protective against muscle atrophy and oxidative cell death. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1643-1658, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27034   open full text
• Circular RNA circRNA_15698 aggravates the extracellular matrix of diabetic nephropathy mesangial cells via miR‐185/TGF‐β1.
  Wei Hu, Qing Han, Lei Zhao, Li Wang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- circRNA_15698/miR‐185/TGF‐β1 promoted the extracellular matrix (ECM) related proteins synthesis. Circular RNAs (circRNAs) are a novel type of noncoding RNAs that modulate the pathogenesis of multiple diseases. Nevertheless, the role of circRNAs in diabetic nephropathy (DN) pathogenesis is still ambiguous. In the current study, our team aims to investigate the expression profiles of circRNAs in DN and identify the function of circRNA on mesangial cells. CircRNAs microarray analysis revealed dysregulated circRNA in db/db DN mice, and circRNA_15698 was validated to be upregulated in both db/db mice and mouse mesangial cells (SV40‐MES13) that were exposed to high glucose (25 mM) using real‐time polymerase chain reaction. Loss‐of‐functional experiments showed that circRNA_15698 knockdown significantly inhibited the expression levels of collagen type I (Col. I), collagen type IV (Col. IV), and fibronectin. Moreover, the cellular localization of circRNA_15698 was mainly in the cytoplasm. Bioinformatics tools and luciferase reporter assay confirmed that circRNA_15698 acted as a ‘sponge’ of miR‐185, and then positively regulated the transforming growth factor‐β1 (TGF‐β1) protein expression, suggesting a circRNA_15698/miR‐185/TGF‐β1 pathway. Further validation experiments validated that circRNA_15698/miR‐185/TGF‐β1 promoted extracellular matrix (ECM)‐related protein synthesis. In summary, our study preliminarily investigates the role of circRNAs in mesangial cells and ECM accumulation, providing a novel insight for DN pathogenesis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1469-1476, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26959   open full text
• Nine hub genes as the potential indicator for the clinical outcome of diabetic nephropathy.
  Xiaoping Song, Min Gong, Yanping Chen, Hui Liu, Jun Zhang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this study, a total of nine genes were found to have an important function in the occurrence and development of DN, among which six were reported earlier, and the result was further verified in this study. More important, the three remaining genes, VAV1, LCK, and PLK1, were identified as novel and had the potential to serve as the predictive indicator in the diagnosis and treatment of DN, which still needs further research. Diabetic nephropathy (DN) is one of the most serious and dangerous chronic complications caused by diabetes mellitus, and the identification and development of novel biomarkers could be beneficial for the diagnosis and prognosis of DN patients. This study focused on researching the differently expressed pattern of the DN samples from glomeruli and tubulointerstitium. Significance analysis of microarrays (SAM) was used to identify differentially regulated genes in 44 microdissected human kidney samples. Functional enrichment analysis was used to analyze the functions these genes are mostly enriched in. Besides, protein–protein interaction (PPI) network was used to select the hub genes that were associated with DN. The gene expression pattern of DN samples from glomeruli and tubulointerstitium was found to be quite different. It showed that the recurrence rate of DN originating from glomeruli and control samples was lower than that from tubulointerstitium and control samples. A total of 332 differentially expressed genes were identified between glomeruli tissues and tubulointerstitium tissues. PPI network analysis was performed on these 332 genes. Finally, nine hub genes were selected as the most potential biomarkers in the occurrence of DN. In conclusion, a total of 332 genes were found to be related to DN, and these genes were found to be mostly enriched in pathways such as immunity, inflammatory, and vascular pathways. Three genes VAV1, LCK, and Plk had the potential to serve as indicators for the occurrence and development of DN in clinical management. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1461-1468, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26958   open full text
• Downregulated microRNA‐488 enhances odontoblast differentiation of human dental pulp stem cells via activation of the p38 MAPK signaling pathway.
  Dan Yu, Xue Zhao, Jin‐Zhang Cheng, Di Wang, Hui‐Hui Zhang, Guang‐Hong Han.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Downregulation of miR‐488 promotes odontoblastic differentiation of human dental pulp stem cells (hDPSCs) by targeting mitogen‐activated protein kinases 1 (MAPK1) through the p38 MAPK signaling pathway, paving the basis for further study about hDPSCs. Human dental pulp stem cells (hDPSCs) are primarily derived from the pulp tissues of permanent third molar teeth. They were widely used in human bone tissue engineering. It was previously indicated that microRNA (miR) expressions are closely associated with hDPSCs development. However, the specific effect of miR‐488 on hDPSCs still remains unclear. In this study, we aimed to investigate effects of miR‐488 on the differentiation of hDPSCs into odontoblast cells through the p38 mitogen‐activated protein kinases (MAPK) signaling pathway by binding to MAPK1. The hDPSCs were isolated and cultured in vitro. Dual‐luciferase reporter gene assay was performed to test the relationship between MAPK1 (p38) and miR‐488. Reverse transcription quantitative polymerase chain reaction and western blot analysis were used to detect the mRNA and protein expressions of p38 MAPK signaling pathway‐related genes (MAPK1, Ras, and Mitogen‐activated protein kinase kinase 3/6 [MKK3/6]), along with expressions of dentin Sialophosphoprotein (DSPP), alkaline phosphatase (ALP), and osteonectin (OCN). ALP staining and alizarin red staining were conducted to detect ALP activity and degree of mineralization. Initially, we found that MAPK1 was the target gene of miR‐488. Besides, downregulation of miR‐488 was observed to stimulate the p38 MAPK signaling pathway and to increase the messenger RNA and protein expressions of DSPP, ALP, and OCN. Furthermore, ALP activity and formation of a mineralized nodule in hDPSCs were enhanced upon downregulation of miR‐488. The aforementioned findings provided evidence supporting that downregulation of miR‐488 promotes odontoblastic differentiation of hDPSCs through the p38 MAPK signaling pathway by targeting MAPK1, paving the basis for further study about hDPSCs. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1442-1451, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26950   open full text
• Preparation of superparamagnetic iron oxide/doxorubicin loaded chitosan nanoparticles as a promising glioblastoma theranostic tool.
  Leila Gholami, Mohsen Tafaghodi, Bita Abbasi, Majid Daroudi, Reza Kazemi Oskuee.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Illustration of ACSD nanoparticles (NPs) preparation by the ionic gelation method, and subsequent hypothetical scheme of ACSD NPs internalization. Theranostic nanoparticles (NPs) are promising for opening new windows toward personalized disease management. Using a single particle capable of both diagnosis and drug delivery, is the major benefit of such particles. In the present study, chitosan NPs were used as a dual action carrier for doxorubicin (DOX; chemotherapeutic agent) and superparamagnetic iron oxide nanoparticles (SPIONs; imaging agent). SPIONs and DOX were loaded at different concentrations within poly‐l‐arginine‐chitosan‐triphosphate matrix (ACSD) using the ionic gelation method. NPs’ size were in the range of 184.33 ± 4.4 nm. Drug release analysis of DOX loaded NPs (NP‐DOX) showed burst release at pH 5.5 (as in tumor environment) and slow release at pH 7.4 (physiological condition), demonstrating pH‐sensitive drug release profile. NP‐DOX internalization was confirmed by flowcytometry and fluorescent microscopy. Uptake process results were corroborated by accumulation of drug in the intracellular space. Iron content was evaluated by inductively coupled plasma and prussian blue staining. In vitro magnetic resonance imaging (MRI) showed a decline in T 2 relaxation times by increasing iron concentration. MRI analysis also confirmed uptake of NPs at the optimum concentration in C6 glioma cells. In conclusion, ACSD NPs could be utilized as a promising theranostic formulation for both diagnosis and treatment of glioblastoma. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1547-1559, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27019   open full text
• TBX18 transcription factor overexpression in human‐induced pluripotent stem cells increases their differentiation into pacemaker‐like cells.
  Armita M. Gorabi, Saeideh Hajighasemi, Hossein A. Tafti, Amir Atashi, Masoud Soleimani, Nasser Aghdami, Ali K. Saeid, Vahid Khori, Yunes Panahi, Amirhossein Sahebkar.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- In this in vitro study, the effect of T‐box 18 gene expression in human‐induced pluripotent‐stem‐cell‐derived cardiomyocytes to induce pacemaker‐like cells was examined. Background The discovery of gene‐ and cell‐based strategies has opened a new area to investigate novel approaches for the treatment of many conditions caused by cardiac cell failure. The TBX18 (T‐box 18) transcription factor is considered as a prominent factor in the sinoatrial node (SAN) formation during the embryonic development. In this in vitro study, the effect of TBX18 gene expression on human‐induced pluripotent‐stem‐cell‐derived cardiomyocytes (hiPS‐CMs) to induce pacemaker‐like cells was examined. Methods The human‐dermal‐fibroblast‐derived iPSCs were transfected using chemical, physical, and Lentiviral methods of TBX18 gene delivery during differentiation into cardiomyocytes (CMs). After the differentiation process through small‐molecule‐based temporal modulation of the Wnt signaling pathway, the hiPSC‐CMs were analyzed using the real‐time polymerase chain reaction, immunocytochemistry, immunofluorescence, whole‐cell patch‐clamp recording, and western blotting to investigate the accuracy of differentiation and identify the effect exerted by TBX18. Results The hiPS‐CMs showed spontaneous beating and expressed specific markers of cardiac cells. The lentiviral‐mediated TBX18 delivery was the most efficient method for transfection. The results showed the increment in Connexin 43 expression among untransfected hiPS‐CMs, whereas this protein was significantly downregulated followed by TBX18 overexpression. TBX18‐hiPSCMs were detected with pacemaker cell features. Conclusions It was demonstrated that the TBX18 gene is able to conduct hiPSCs to differentiate into pacemaker‐like cells. The TBX18 gene delivery seems to have the potential for the development of biological pacemakers; however, more investigations are still needed to assess its usefulness to fix arrhythmic conditions with SAN failure basis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1534-1546, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27018   open full text
• ESM‐1 promotes adhesion between monocytes and endothelial cells under intermittent hypoxia.
  Haili Sun, Huina Zhang, Kun Li, Hao Wu, Xiaojun Zhan, Fang Fang, Yanwen Qin, Yongxiang Wei.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Intermittent hypoxia (IH), the key property of obstructive sleep apnea, is closely associated with endothelial dysfunction. Endothelial‐cell‐specific molecule‐1 (ESM‐1, Endocan) is a novel, reported molecule linked to endothelial dysfunction. The aim of this study is to evaluate the effect of IH on ESM‐1 expression and the role of ESM‐1 in endothelial dysfunction. ESM‐1 is significantly upregulated by HIF‐1α/vascular endothelial growth factor pathway under IH in endothelial cells, playing a role in enhancing adhesion between monocytes and endothelial cells, which might be a potential target for IH‐induced endothelial dysfunction. Intermittent hypoxia (IH), the key property of obstructive sleep apnea (OSA), is closely associated with endothelial dysfunction. Endothelial‐cell‐specific molecule‐1 (ESM‐1, Endocan) is a novel, reported molecule linked to endothelial dysfunction. The aim of this study is to evaluate the effect of IH on ESM‐1 expression and the role of ESM‐1 in endothelial dysfunction. We found that serum concentration of ESM‐1, inter‐cellular adhesion molecule‐1 (ICAM‐1), and vascular cell adhesion molecule‐1 (VCAM‐1) is significantly higher in patients with OSA than healthy volunteers (p < 0.01). The expression of ESM‐1, hypoxia‐inducible factor‐1 alpha (HIF‐1α), and vascular endothelial growth factor (VEGF) was significantly increased in human umbilical vein endothelial cells (HUVECs) by treated IH in a time‐dependent manner. HIF‐1α short hairpin RNA and vascular endothelial growth factor receptor (VEGFR) inhibitor inhibited the expression of ESM‐1 in HUVECs. ICAM‐1 and VCAM‐1 expressions were significantly enhanced under IH status, accompanied by increased monocyte–endothelial cell adhesion rate ( p < 0.001). Accordingly, ESM‐1 silencing decreased the expression of ICAM‐1 and VCAM‐1 in HUVECs, whereas ESM‐1 treatment significantly enhanced ICAM‐1 expression accompanied by increasing adhesion ability. ESM‐1 is significantly upregulated by the HIF‐1α/VEGF pathway under IH in endothelial cells, playing a critical role in enhancing adhesion between monocytes and endothelial cells, which might be a potential target for IH‐induced endothelial dysfunction. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1512-1521, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27016   open full text
• Exosome‐encapsulated miR‐6089 regulates inflammatory response via targeting TLR4.
  Donghua Xu, Meiyan Song, Chunxiang Chai, Jinghua Wang, Chengwen Jin, Xiaodong Wang, Min Cheng, Shushan Yan.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- TLR4 is a direct target for miR‐6089. MiR‐6089 regulates the generation of IL‐6, IL‐29, and TNF‐α by targetedly controlling the TLR4 signaling. Exosome‐encapsulated microRNAs (miRNAs) have been identified as potential biomarkers in autoimmune diseases. However, little is known about the role of exosome‐delivered miRNAs in rheumatoid arthritis (RA). In this study, we investigated the profile of specific exosomal miRNAs by microarray analysis of serum exosomes from three patients with RA and three healthy controls. Quantitative real‐time PCR (qRT‐PCR) was performed to validate the aberrantly expressed exosomal miRNAs. A total of 20 exosome‐encapsulated miRNAs were identified to be differently expressed in the serum of patients with RA compared with controls. Interestingly, we found that exosome‐encapsulated miR‐6089 was significantly decreased after validation by qRT‐PCR in serum exosomes from 76 patients with RA and 20 controls. Besides, miR‐6089 could inhibit lipopolysaccharide (LPS)‐induced cell proliferation and activation of macrophage‐like THP‐1 cells. TLR4 was a direct target for miR‐6089. MiR‐6089 regulated the generation of IL‐6, IL‐29, and TNF‐α by targetedly controlling TLR4 signaling. In conclusion, exosome‐encapsulated miR‐6089 regulates LPS/TLR4‐mediated inflammatory response, which may serve as a novel, promising biomarker in RA. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1502-1511, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27014   open full text
• Sex differences in murine cardiac pathophysiology with hyperoxia exposure.
  Jennifer L. Rodgers, Lydia E. Rodgers, Zhi Tian, Diane Allen‐Gipson, Siva K. Panguluri.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Hyperoxia is a common intervention in ICU. It is known that sex differences do exist in cardiovascular physiology, and this manuscript describes the sex differences in hyperoxia induced cardiac pathophysiology. Hyperoxia (>90% oxygen) is commonly implemented in mechanically ventilated patients. Reports suggest that hyperoxia is directly associated with in‐hospital mortality in ventilated patients. Certain studies also show that mortality in women undergoing mechanical ventilation is significantly higher than that in men. Additionally, females are predisposed to certain cardiac electrophysiological risks, including QTc prolongation. In this study, we assessed the impact of hyperoxia in male and female mice (C57BL/6J) at age 8–10 weeks. On completion of either hyperoxia or normoxia exposures, physical, hemodynamic, biochemical, functional, electrophysiological, and molecular assessments were conducted. Hyperoxia‐exposed mice lost a significant amount of body mass, compared with normoxia controls, in both sexes. However, while both genders developed brady‐arrhythmia after hyperoxia exposure, female mice exhibited significantly reduced heart rates compared with males, with significantly elevated RR intervals. Additionally, 50% mortality was observed in females, whereas no mortality was reported in males. Furthermore, unlike in male mice, we observed no hypertrophy upon hyperoxia exposure in female mice. We reported that both hyperoxia‐treated male and female mice exhibit significant hyperdynamic left ventricular ejection fraction, which is marked by % ejection fraction > 70 compared with the normoxia controls. We also noted significant reductions in stroke volume and cardiac output in both mice with hyperoxia. Surface ECG also demonstrated that hyperoxia exposure significantly augments RR, PR, QRS, QTc, and JT intervals in both sexes. Molecular analysis of left ventricular tissue demonstrated dysregulation of potassium ion channels in hyperoxia‐treated males and females. In summary, we determined that sex differences are present with 72 hr hyperoxia exposure. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1491-1501, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27010   open full text
• Inhibitory role of large intergenic noncoding RNA‐ROR on tamoxifen resistance in the endocrine therapy of breast cancer by regulating the PI3K/Akt/mTOR signaling pathway.
  Peng‐Wei Lu, Lin Li, Fang Wang, Yuan‐Ting Gu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Our study demonstrated that silencing lincRNA‐ROR could increase the sensitivity of BC MAD‐MB‐231 cells to TAM by suppressing the activation of P13K/Akt/mTOR signaling pathway. Breast cancer (BC) is the second‐leading cause of central nervous system metastases among severe malignancies. This study aimed at investigating the underlying mechanism by which large intergenic noncoding RNA‐regulator of reprogramming (lincRNA‐ROR) affects the tamoxifen (TAM) resistance of BC cells by regulating the PI3K/Akt/mTOR signaling pathway. Immortalized human mammary epithelial cell line (MCF10A) and BC cell lines (MCF‐7, MDA‐MB‐231, T47D, BCAP‐37, and ZK‐75‐1) were cultured, and BC tissues and adjacent normal breast tissues were collected from 152 BC patients. LincRNA‐ROR expression in tissues and cells were detected using reverse transcription quantitative polymerase chain reaction. RNA interference was used to silence lincRNA‐ROR in MDA‐MB‐231 cells, and then the cell proliferation and apoptosis were detected by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay and annexin‐V and propidium iodide (PI) double staining respectively. The expression of apoptosis‐related proteins and PI3K/Akt/mTOR signaling pathway–related proteins was measured by performing western blot assay. The BC tissues and cells presented a higher expression of lincRNA‐ROR. MAD‐MB‐231 cells exhibited the highest lincRNA‐ROR expression. After lincRNA‐ROR silencing, MAD‐MB‐231 cells showed decreased proliferation, and increased sensitivity to TAM. Besides, the apoptosis‐promoting effect of TAM on MAN‐MB‐231 cells significantly increased. The expression of PI3K/Akt/mTOR signaling pathway–related proteins and the PI3K/Akt/mTOR signaling pathway were repressed by TAM after silencing lincRNA‐ROR. Our study demonstrated that silencing lincRNA‐ROR could increase the sensitivity of BC MAD‐MB‐231 cells to TAM by suppressing the activation of P13K/Akt/mTOR signaling pathway. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1904-1912, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27066   open full text
• Anticancer effects of echinacoside in hepatocellular carcinoma mouse model and HepG2 cells.
  Ying Ye, Yanan Song, Juhua Zhuang, Guoyu Wang, Jing Ni, Wei Xia.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Echinacoside (ECH) could attenuate diethylnitrosamine‐induced hepatocellular carcinoma (HCC) in mice, and exerted antiproliferative and proapoptotic functions on HepG2 HCC cell line. ECH exerted its antitumor activity via decreasing TREM2 expression and phosphoinositide 3‐kinase/AKT signaling. Echinacoside (ECH) is a phenylethanoid glycoside extracted from a Chinese herbal medicine, Cistanches salsa. ECH possesses many biological properties, including anti‐inflammation, neural protection, liver protection, and antitumor. In the current study, we aimed to explore the effects of ECH on hepatocellular carcinoma (HCC) and the underlying mechanisms. The results showed that ECH could attenuate diethylnitrosamine (DEN)‐induced HCC in mice, and exerted antiproliferative and proapoptotic functions on HepG2 HCC cell line. ECH exposure in HepG2 cells dose‐dependently reduced the phosphorylation of AKT (p‐AKT) and enhanced the expression of p21 (a cell cycle inhibitor) and Bax (a proapoptotic protein). Furthermore, ECH significantly suppressed insulin‐like growth factor‐1‐induced p‐AKT and cell proliferation. These data indicated that phosphoinositide 3‐kinase (PI3K)/AKT signaling was involved in the anti‐HCC activity of ECH. Gene set enrichment analysis results revealed a positive correlation between the PI3K pathway and triggering receptors expressed on myeloid cells 2 (TREM2) expression in HCC tissues. ECH exposure significantly decreased TREM2 protein levels in HepG2 cells and DEN‐induced HCC. Furthermore, ECH‐mediated proliferation inhibition and AKT signaling inactivation were notably attenuated by TREM2 overexpression. In conclusion, ECH exerted its antitumor activity via decreasing TREM2 expression and PI3K/AKT signaling. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1880-1888, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27063   open full text
• Chibby is a weak regulator of β‐catenin activity in gastric epithelium.
  Feng‐Qian Li, Luis Chiriboga, Margaret A. Black, Ken‐Ichi Takemaru, Robert D. Raffaniello.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Chibby (Cby) is a 14‐kDa protein that inhibits β‐catenin localization to the nucleus and represses β‐catenin‐induced transcriptional activity. Cby expression was decreased in gastric cancer (GC) tissue and Cby expression altered β‐catenin localization in cultured GC cells. However, Cby did not affect cell proliferation rates or β‐catenin‐induced protein expression. The canonical Wnt–β‐catenin pathway is important in normal development. Mutations in β‐catenin or proteins involved with regulating its phosphorylation or localization result in its nuclear accumulation where it activates its target genes and stimulates cell proliferation. This pathway is dysregulated in many different types of cancer, including gastric cancer (GC). Chibby (Cby) is a 14‐kDa protein that inhibits β‐catenin localization to the nucleus and represses β‐catenin‐induced transcriptional activity. In the current study, we examined the expression and function of Cby in normal and cancerous human gastric tissue. Reverse‐transcription polymerase chain reaction and immunohistochemistry revealed that Cby is expressed in human stomach and localized to glandular elements. Immunohistochemical staining intensity of Cby was decreased in GC tissue when compared with normal gastric epithelium. In AGS cells, a human gastric carcinoma cell line, Cby expression was low. Stable AGS cell transfectants overexpressing Cby were prepared. Cby overexpression did not affect proliferation rates or β‐catenin levels. However, confocal microscopy and subcellular fractionation studies revealed that Cby overexpression resulted in a small decrease in nuclear β‐catenin. Moreover, Cby overexpression caused a molecular weight shift in nuclear β‐catenin and resulted in decreased β‐catenin signaling in AGS cells as measured by the TopFlash assay. However, Cby overexpression did not affect c‐Myc protein levels. To conclude, Cby expression was decreased in GC samples and Cby expression altered β‐catenin localization in cultured GC cells. However, Cby did not affect cell proliferation rates or β‐catenin‐induced protein expression. Cby may be involved in the early events in the pathogenesis of GC. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1871-1879, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27062   open full text
• Circulating exosomal CPNE3 as a diagnostic and prognostic biomarker for colorectal cancer.
  Bo Sun, Yiming Li, Yiming Zhou, Tien Khee Ng, Chao Zhao, Qiaoqiang Gan, Xiaodong Gu, Jianbin Xiang.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Exosomal Copine III (CPNE3) levels are associated with the tumor extent and may serve as a diagnostic biomarker in patients with colorectal cancer (CRC). Combining measurement of exosomal CPNE3 in plasma with carcinoembryonic antigen (CEA) may further improve the diagnostic accuracy. The high exosomal CPNE3 level was a reliable indicator for worse disease‐free survival (DFS) and overall survival (OS). Exosomal proteins are emerging as relevant diagnostic and prognostic biomarkers for cancer. This study was aimed at illustrating the clinical significance of exosomal Copine III (CPNE3) purified from the plasma of colorectal cancer (CRC) patients. The CPNE3 expression levels in CRC tissues were analyzed by real‐time PCR, western blot, and immunohistochemistry. Plasma exosomes were isolated to examine the CPNE3 level using ELISA. Pearson’s correlation analysis was performed to investigate the CPNE3 levels between CRC tissues and matched plasma samples. Receiver operating characteristic curve analysis was developed to measure the diagnostic performance of exosomal CPNE3. The Kaplan–Meier method and Cox's proportional hazards model were utilized to determine statistical differences in survival times. CPNE3 showed increased expressions in the CRC tissues. A moderately significant correlation was found between CPNE3 expression in CRC tissues by immunohistochemistry and matched serum exosomal CPNE3 expression by ELISA (r = 0.645,(r = 0.645, p < 0.001). < 0.001). Exosomal CPNE3 yielded a sensitivity of 67.5% and a specificity of 84.4% in CRC at the cutoff value of 0.143 pg per 1ug1 ug exosome. Combined data from carcinoembryonic antigen and exosomal CPNE3 achieved 84.8% sensitivity and 81.2% specificity as a diagnostic tool. CRC patients with lower exosomal CPNE3 levels had substantially better disease‐free survival (hazard ratio [HR], 2.9; 95% confidence interval [CI]: 1.3–6.4; p = 0.009) = 0.009) and overall survival (HR, 3.4; 95% CI: 1.2–9.9; p = 0.026) = 0.026) compared with those with higher exosomal CPNE3 levels. Exosomal CPNE3 show potential implications in CRC diagnosis and prognosis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1416-1425, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26936   open full text
• Venom of the Phoneutria nigriventer spider alters the cell cycle, viability, and migration of cancer cells.
  Natália Barreto dos Santos, Amanda Pires Bonfanti, Thomaz Augusto Alves da Rocha‐e‐Silva, Pedro Ismael da Silva, Maria Alice da Cruz‐Höfling, Liana Verinaud, Catarina Rapôso.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Phoneutria nigriventer venom (PnV) decreases viability of brain and cervix tumor, but not of nontumor cells. PnV decreases the viability of glioma cells primarily by modulating the cell cycle, while the viability of glioblastoma cells is reduced mainly by inducing death. PnV delays migration of tumor cells and decreases invasiveness. The mechanisms of cancer involve changes in multiple biological pathways. Multitarget molecules, which are components of animal venoms, are therefore a potential strategy for treating tumors. The objective of this study was to screen the effects of Phoneutria nigriventer spider venom (PnV) on tumor cell lines. Cultured human glioma (NG97), glioblastoma (U‐251) and cervix adenocarcinoma (HeLa) cells, and nontumor mouse fibroblasts (L929) were treated with low (14 µg/ml) and high (280 µg/ml) concentrations of PnV, and analyzed through assays for cell viability (thiazolyl blue tetrazolium blue), proliferation (carboxyfluorescein succinimidyl ester), death (annexin V/propidium iodide [Pi]), the cell cycle (Pi), and migration (wound healing and transwell assay). The venom decreased the viability of U‐251 cells, primarily by inducing cell death, and reduced the viability of NG97 cells, primarily by inhibiting the cell cycle. The migration of all the tumor cell lines was delayed when treated with venom. The venom significantly affected all the tumor cell lines studied, with no cytotoxic effect on normal cells (L929), although the nonglial tumor cell (HeLa) was less sensitive to PnV. The results of the current study suggest that PnV may be composed of peptides that are highly specific for the multiple targets involved in the hallmarks of cancer. Experiments are underway to identify these molecules. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1398-1415, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26935   open full text
• Nicotine downregulates microRNA‐200c to promote metastasis and the epithelial–mesenchymal transition in human colorectal cancer cells.
  Zhou Lei, Yang Xiaomin, Huang He, Chen Jian, Xu Xiaowu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2 1) Cigarette smoking was independently associated with poorer survival, regardless of American Joint Committee on Cancer stage. 2) Nicotine progressively decreased microRNA (miR)‐200c expression in a time‐ and dose‐dependent manner in colorectal cancer (CRC) cells. 3) Nicotine promoted proliferation, migration, and invasion and the epithelial–mesenchymal transition in CRC cells, though overexpression of miR‐200c abrogated these nicotine‐induced effects. Background Cigarette smoking is the most well‐established risk factor for colorectal cancer (CRC). However, the mechanisms of smoking‐associated colorectal carcinogenesis are poorly understood. Methods The effects of prediagnosis tobacco use on clinical characteristics, overall survival (OS), and recurrence‐free survival (RFS) were analyzed in 396 patients with CRC. Associations between smoking status and OS and RFS were evaluated using Cox’s proportional hazards regression. Furthermore, the effects of nicotine on the CRC cell lines SW620 and HT‐29 were evaluated using in vitro assays. Results “Ever smoking” was associated with elevated serum carcinoembryonic antigen, American Joint Committee on Cancer T category, metastasis, and poorer OS and RFS in patients with CRC (OS: hazard ratio [HR] = 1.74, 95% confidence interval [CI], 1.07–2.81, p = 0.025; RFS: HR = 1.66, 95% CI: 1.18–2.34, p = 0.004). MicroRNA (miR)‐200c was downregulated in CRC and tumor‐adjacent tissues from ever smokers compared with the corresponding tissues from never smokers with CRC. Nicotine inhibited miR‐200c expression in a dose‐ and time‐dependent manner in SW620 and HT‐29 CRC cell lines. Nicotine induced cell proliferation, migration, and invasion and promoted the epithelial–mesenchymal transition in SW620 and HT‐29 cells, and these effects were attenuated by overexpression of miR‐200c. Conclusion Our findings support the adverse effects of prediagnosis cigarette smoking on prognosis and clinical behavior in CRC. We demonstrate a novel oncogenic mechanism by which nicotine promotes growth and metastasis in CRC by downregulating miR‐200c. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1369-1379, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.26933   open full text
• Rosmarinic acid inhibits nicotine‐induced C‐reactive protein generation by inhibiting NLRP3 inflammasome activation in smooth muscle cells.
  Yang Yao, Junjun Mao, Shouzhu Xu, Lei Zhao, Lihui Long, Lin Chen, Dongmin Li, Shemin Lu.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- (1) Rosmarinic acid plays a protective role in nicotine‐induced atherosclerosis. (2) Rosmarinic acid downregulates nicotine‐induced C‐reactive protein expression by inhibiting ROS‐mediated NLRP3 inflammasome activation in rat vascular smooth muscle cells. Atherosclerosis is widely known to be a chronic inflammatory disease. C‐reactive protein (CRP), an important inflammatory factor, plays an essential role in the pathogenesis of atherosclerosis. Nicotine, the main addictive component of cigarette, has been shown to induce the production of CRP. The aim of this study was to investigate the effect of rosmarinic acid (RA), a polyphenol with antiinflammatory activity, on nicotine‐induced elevation of CRP in vascular smooth muscle cells (VSMCs). We found that pretreatment of VSMCs with RA attenuated nicotine‐induced expression of CRP in a time‐ and dose‐dependant manner. In addition, RA also inhibited the activation of NLR family pyrin domain containing 3 (NLRP3) inflammasome and reactive oxygen species (ROS) production resulting from nicotine treatment in VSMCs. To confirm these findings in vivo, we constructed a nicotine‐induced atherosclerosis rat model. RA did not significantly reduce the serum nicotine level of the rats, whereas it significantly decreased the levels of serum lipids, including concentrations of cholesterol, triglycerides, and low‐density lipoprotein cholesterol, and the serum level of CRP. RA also led to diminished nicotine‐induced activation of NLRP3 inflammasome and elevation in the CRP level in the aortic tissue of the model rats. The results of this study suggested a protective role of RA in nicotine‐induced atherosclerosis by inhibiting the ROS–NLRP3 inflammasome–CRP axial, and RA therefore represented a potential effective therapeutic approach to atherosclerosis, in particular for those who smoke. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1758-1767, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27046   open full text
• Ameloblastin attenuates RANKL‐mediated osteoclastogenesis by suppressing activation of nuclear factor of activated T‐cell cytoplasmic 1 (NFATc1).
  Wichida Chaweewannakorn, Wataru Ariyoshi, Toshinori Okinaga, Yuko Fujita, Kenshi Maki, Tatsuji Nishihara.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Ameloblastin (Ambn) suppresses receptor activator of nuclear factor kappa‐B ligand (RANKL)‐induced osteoclastogenesis in three specific pathways; inhibition of JNK and p38 mitogen‐activated protein kinase (MAPK)‐mediated c‐Fos activation, calcium oscillation and cAMP‐response element‐binding protein (CREB)‐mediated nuclear factor of activated T cells cytoplasmic 1 (NFATc1) upregulation, and B‐lymphocyte‐induced maturation protein‐1 (Blimp1)‐mediated negative regulation of osteoclastogenesis Ameloblastin (Ambn) is an extracellular matrix protein and member of the family of enamel‐related gene products. Like amelogenin, Ambn is mainly associated with tooth development, especially biomineralization of enamel. Previous studies have shown reductions in the skeletal dimensions of Ambn‐deficient mice, suggesting that the protein also has effects on the differentiation of osteoblasts and/or osteoclasts. However, the specific pathways used by Ambn to influence osteoclast differentiation have yet to be identified. In the present study, two cellular models, one based on bone marrow cells and another on RAW264.7 cells, were used to examine the effects of Ambn on receptor activator of nuclear factor kappa‐B ligand (RANKL)‐induced osteoclastogenesis. The results showed that Ambn suppresses osteoclast differentiation, cytoskeletal organization, and osteoclast function by the downregulation of the number of tartrate‐resistant acid phosphatase (TRAP)‐positive multinucleated osteoclasts, actin ring formation, and areas of pit resorption. The expression of the osteoclast‐specific genes TRAP, MMP9, cathepsin K, and osteoclast stimulatory transmembrane protein (OC‐STAMP) was abolished in the presence of Ambn, while that of nuclear factor of activated T cells cytoplasmic 1 (NFATc1), the master regulatory factor of osteoclastogenesis, was also attenuated by the downregulation of c‐Fos expression. In Ambn‐induced RAW264.7 cells, phosphorylation of cAMP‐response element‐binding protein (CREB), c‐Jun N‐terminal kinase (JNK), and p38 mitogen‐activated protein kinase (p38 MAPK), but not extracellular signal‐regulated kinase 1/2 (ERK1/2), was reduced. Calcium oscillation was also decreased in the presence of Ambn, suggesting its involvement in both RANKL‐induced osteoclastogenesis and costimulatory signaling. B‐lymphocyte‐induced maturation protein‐1 (Blimp1), a transcriptional repressor of negative regulators of osteoclastogenesis, was also downregulated by Ambn, resulting in the elevated expression of v‐maf musculoaponeurotic fibrosarcoma oncogene family, protein B (MafB), B‐cell lymphoma 6 (Bcl6), and interferon regulatory factor‐8 (Irf8). Taken together, these findings suggest that Ambn suppresses RANKL‐induced osteoclastogenesis by modulating the NFATc1 axis. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1745-1757, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27045   open full text
• CircHIPK3 promotes proliferation and invasion in nasopharyngeal carcinoma by abrogating miR‐4288‐induced ELF3 inhibition.
  Zhaoyang Ke, Fei Xie, Chaopan Zheng, Daishi Chen.
  Journal of Cellular Physiology. November 20, 2018
  --- - |2- Our findings demonstrated that circular homeodomain interacting protein kinase 3 (circHIPK3) facilitated nasopharyngeal carcinoma (NPC) progression through protecting E74‐like ETS transcription factor 3 (ELF3) from microRNA (miR)‐4288‐mediated silencing, which suggested that the circHIPK3‐miR‐4288‐ELF3 regulatory loop might be a potential target for NPC prevention. Circular RNAs (circRNAs) are reported to regulate the development and progression of multiple cancers. However, the functions of circRNAs in nasopharyngeal carcinoma (NPC) are unclear. In this study, we identified that circular homeodomain interacting protein kinase 3 (circHIPK3) was highly expressed in NPC tissues and cell lines. Moreover, we found that circHIPK3 expression levels could act as a prognostic marker in NPC patients. We showed that circHIPK3 silence repressed NPC cell proliferation, migration, and invasion in vitro. In addition, circHIPK3 depletion dramatically repressed tumor growth and metastasis in vivo. Mechanistically, we revealed circHIPK3 as a competing endogenous RNA of microRNA (miR)‐4288 that targets E74‐like ETS transcription factor 3 (ELF3) in NPC cells. We found that miR‐4288 inhibition reversed the effects of circHIPK3 silence on NPC cells. Furthermore, rescue assays also indicated that circHIPK3 promoted the malignant behaviors of NPC cells via enhancing ELF3 expression by suppressing the miR‐4288 levels. In conclusion, our findings demonstrated that circHIPK3 facilitated NPC progression through protecting ELF3 from miR‐4288‐mediated silencing, which suggested that the circHIPK3‐miR‐4288‐ELF3 regulatory loop might be a potential target for NPC prevention. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1699-1706, February 2019. '

  November 20, 2018   doi: 10.1002/jcp.27041   open full text
• Prognostic value of pretreatment systemic immune‐inflammation index in patients with gastrointestinal cancers.
  Yi Zhang, Shibu Lin, Xianjin Yang, Rong Wang, Lingyan Luo.
  Journal of Cellular Physiology. October 24, 2018
  --- - |2 Abstract Background Numerous studies have reported the relationship between systemic immune‐inflammation index (SII) and prognosis in gastrointestinal (GI) cancers, but no consensus has been reached. We aimed to systematically evaluate the prognostic value of SII in patients with GI cancers. Methods Relevant published papers regarding the prognostic value of SII in patients with GI cancers were obtained from a number of electronic databases. The overall hazard ratios and the corresponding 95% confidence intervals (95% CIs) were calculated using a fixed or random effects model to assess the relationship between SII and prognosis through Stata SE 12.0. Results A total of 24 eligible published articles with 9,626 patients were included. From the pooled results, we found that high SII indicated worse overall survival (OS) in patients with GI cancers (HR = 1.52, 95%CI: 1.29–1.74). And patients with high SII had poorer disease‐free survival (HR: 2.28, 95% CI: 1.46–3.10), time to recurrence (HR: 1.70, 95% CI: 1.11–2.30), and recurrence‐free survival (HR: 1.60, 95% CI: 1.19–2.00) when compared with those with low SII values. Conclusions SII might serve as a noninvasive and powerful tool for predicting survival outcome in patients with GI cancers. - Journal of Cellular Physiology, EarlyView.
  

  October 24, 2018   doi: 10.1002/jcp.27373   open full text
• Thrombin cleavage of osteopontin controls activation of hepatic stellate cells and is essential for liver fibrogenesis.
  Guangying Cui, Jianing Chen, Zhongwen Wu, Haijun Huang, Lin Wang, Yan Liang, Ping Zeng, Jiezuan Yang, Toshimitsu Uede, Hongyan Diao.
  Journal of Cellular Physiology. October 23, 2018
  --- - |2- Abstract Liver biopsy is the current reliable way of evaluating liver fibrosis. However, no specific sera biomarker could be applied in clinical diagnosis. As the pivotal role of osteopontin (OPN) reported in numerous liver diseases, thrombin‐cleaved OPN (Thr‐OPN) exposes an integrin‐binding motif that promoted biological functions. Herein, we investigated the potential of Thr‐OPN in liver fibrosis. Using patient samples, mouse models and hepatic stellate cells (HSCs), we analyzed the involvement of Thr‐OPN in liver fibrosis. The result showed that, first, Thr‐OPN level was significantly higher in patients with liver cirrhosis than that in patients with chronic hepatitis B and healthy controls. Thr‐OPN level was positively correlated with liver fibrosis degree in clinical samples. Then in mouse models, it showed a similar correlation between hepatic Thr‐OPN levels and liver fibrosis degree. Thr‐OPN peptides exacerbated liver fibrosis in OPN‐deficient mice, whereas the neutralization of Thr‐OPN alleviated liver fibrosis in wild‐type mice. Furthermore, when compared with full‐length OPN (FL‐OPN), Thr‐OPN exhibited a greater ability to promote HSC activation, proliferation, and migration via mitogen‐activated protein (MAP) kinase and nuclear factor (NF)‐κB pathways. In conclusion, Thr‐OPN, not FL‐OPN, was critically involved in the exacerbation of liver fibrosis by α9 and α4 integrins via MAP kinase and NF‐κB signaling pathway, thus representing a novel diagnostic biomarker and treatment target for liver cirrhosis. - Journal of Cellular Physiology, EarlyView.
  

  October 23, 2018   doi: 10.1002/jcp.27571   open full text
• Tumor suppressors BTG1 and BTG2: Beyond growth control.
  Laurensia Yuniati, Blanca Scheijen, Laurens T. Meer, Frank N. Leeuwen.
  Journal of Cellular Physiology. October 23, 2018
  --- - |2- Abstract Since the identification of B‐cell translocation gene 1 (BTG1) and BTG2 as antiproliferation genes more than two decades ago, their protein products have been implicated in a variety of cellular processes including cell division, DNA repair, transcriptional regulation and messenger RNA stability. In addition to affecting differentiation during development and in the adult, BTG proteins play an important role in maintaining homeostasis under conditions of cellular stress. Genomic profiling of B‐cell leukemia and lymphoma has put BTG1 and BTG2 in the spotlight, since both genes are frequently deleted or mutated in these malignancies, pointing towards a role as tumor suppressors. Moreover, in solid tumors, reduced expression of BTG1 or BTG2 is often correlated with malignant cell behavior and poor treatment outcome. Recent studies have uncovered novel roles for BTG1 and BTG2 in genotoxic and integrated stress responses, as well as during hematopoiesis. This review summarizes what is currently known about the roles of BTG1 and BTG2 in these and other cellular processes. In addition, we will highlight the molecular mechanisms and biological consequences of BTG1 and BTG2 deregulation during cancer progression and elaborate on the potential clinical implications of these findings. - Journal of Cellular Physiology, EarlyView.
  

  October 23, 2018   doi: 10.1002/jcp.27407   open full text
• Growth arrest‐specific 5 attenuates cisplatin‐induced apoptosis in cervical cancer by regulating STAT3 signaling via miR‐21.
  Tingting Yao, Rongbiao Lu, Jun Zhang, Xingyu Fang, Li Fan, Chunxian Huang, Rongchun Lin, Zhongqiu Lin.
  Journal of Cellular Physiology. October 23, 2018
  --- - |2- Abstract Cervical cancer is the most common cause of female cancer‐related mortality worldwide. Decreased expression of long noncoding RNA growth arrest‐specific 5 (GAS5) is found in human cervical cancer tissues and associated with poor prognosis. However, the studies on associations between GAS5 level and malignant phenotypes, as well as sensitivity to chemotherapeutic drug in cervical cancer cells are limited. In this study, overexpression of GAS5 in cervical cancer cells resulted in prohibited cell proliferation and colony formation, which were promoted by siGAS5. Enhanced GAS5 increased cell percentage in the G0/G1 phase and decreased cells percentage in the S phase, whereas reduced expression did not. The malignant behaviors of cervical cancer cells, manifested by cell migration and invasion, could be weakened by the GAS5 overexpression and enhanced by siGAS5. Furthermore, in cisplatin‐induced cell, overexpression of GAS5 reduced cells viability and enhanced apoptosis, whereas in cells transfected with siGAS5, apoptosis eliminated. We have reported the upregulation of microRNA‐21 (miR‐21) and its oncogenetic roles in cervical cancer previously. In this study, we found the negative relationship between the GAS5 and miR‐21. Moreover, the decrease of miR‐21 associated proteins phosphorylated STAT3 and E2F3 was seen in GAS5 overexpressed cells, both of which could be increased by siGAS5. The GAS5 deficiency also reduced miR‐21 target proteins TIMP3 and PDCD4 expressions. Taken together, the GAS5 expression level is inversely associated with malignancy, but positively associated with sensitivity to cisplatin‐induced apoptosis, suggesting that GAS5 could be a biomarker of cisplatin‐resistance in clinical therapy of human cervical cancer. - Journal of Cellular Physiology, EarlyView.
  

  October 23, 2018   doi: 10.1002/jcp.27647   open full text
• GLUT12 expression and regulation in murine small intestine and human Caco‐2 cells.
  Eva Gil‐Iturbe, Rosa Castilla‐Madrigal, Jaione Barrenetxe, Ana Cristina Villaro, María Pilar Lostao.
  Journal of Cellular Physiology. October 23, 2018
  --- - "\nAbstract\nGLUT12 was cloned from the mammary cancer cell line MCF‐7, but its physiological role still needs to be elucidated. To gain more knowledge of GLUT12 function in the intestine, we investigated GLUT12 subcellular localization in the small intestine and its regulation by sugars, hormones, and intracellular mediators in Caco‐2 cells and mice. Immunohistochemical methods were used to determine GLUT12 subcellular localization in human and murine small intestine. Brush border membrane vesicles were isolated for western blot analyses. Functional studies were performed in Caco‐2 cells by measuring α‐methyl‐d‐glucose (αMG) uptake in the absence of sodium. GLUT12 is located in the apical cytoplasm, below the brush border membrane, and in the perinuclear region of murine and human enterocytes. In Caco‐2 cells, GLUT12 translocation to the apical membrane and α‐methyl‐\nd‐glucose uptake by the transporter are stimulated by protons, glucose, insulin, tumor necrosis factor‐α (TNF‐α), protein kinase C, and AMP‐activated protein kinase. In contrast, hypoxia decreases GLUT12 expression in the apical membrane. Upregulation of \nTNF‐α and hypoxia‐inducible factor‐1α (\nHIF‐1α) genes is found in the jejunal mucosa of diet‐induced obese mice. In these animals, GLUT12 expression in the brush border membrane is slightly decreased compared with lean animals. Moreover, an intraperitoneal injection of insulin does not induce GLUT12 translocation to the membrane, as it occurs in lean animals. GLUT12 rapid translocation to the enterocytes’ apical membrane in response to glucose and insulin could be related to GLUT12 participation in sugar absorption during postprandial periods. In obesity, in which insulin sensitivity is reduced, the contribution of GLUT12 to sugar absorption is affected." - Journal of Cellular Physiology, EarlyView.
  

  October 23, 2018   doi: 10.1002/jcp.27231   open full text
• Zic1 mRNA is transiently upregulated in subcutaneous fat of acutely cold‐exposed mice.
  Jessica Perugini, Laura Bordoni, Wiebe Venema, Samantha Acciarini, Saverio Cinti, Rosita Gabbianelli, Antonio Giordano.
  Journal of Cellular Physiology. October 21, 2018
  --- - "\nAbstract\nIn the mammalian adipose organ cold exposure not only activates typical brown adipose tissue, but also induces browning, that is the formation of thermogenic multilocular adipocytes in white, or predominantly white, adipose depots such as subcutaneous fat. Unlike typical brown adipocytes, newly formed thermogenic adipocytes have been reported not to express the gene zinc finger of the cerebellum 1 (Zic1). Here, a time course approach enabled us to document a significant increase in \nZic1 messenger RNA in inguinal subcutaneous fat from acutely (24 hr) cold‐exposed mice, which was paralleled by an increase in multilocular and paucilocular uncoupling protein 1‐positive adipocytes and in parenchymal noradrenergic innervation. This transient, depot‐specific molecular signature was associated not to \nZic1 promoter demethylation, but to chromatin remodeling through an H3K9me3 histone modification. These findings challenge the notion that \nZic1 is exclusively expressed by typical brown adipocytes and suggest its involvement in brown adipocyte precursor differentiation and/or white‐to‐brown adipocyte transdifferentiation." - Journal of Cellular Physiology, EarlyView.
  

  October 21, 2018   doi: 10.1002/jcp.27301   open full text
• Pancreatic endocrine‐like cells differentiated from human umbilical cords Wharton’s jelly mesenchymal stem cells using small molecules.
  Sharath Belame Shivakumar, Dinesh Bharti, Raghavendra Baregundi Subbarao, Ju‐Mi Park, Young‐Bum Son, Imran Ullah, Yong‐Ho Choe, Hyeong‐Jeong Lee, Bong‐Wook Park, Sung‐Lim Lee, Gyu‐Jin Rho.
  Journal of Cellular Physiology. October 21, 2018
  --- - |2- Abstract Following success of pancreatic islet transplantation in the treatment of Type I diabetes mellitus, there is a growing interest in using cell‐based treatment approaches. However, severe shortage of donor islets–pancreas impeded the growth, and made researchers to search for an alternative treatment approaches. In this context, recently, stem cell–based therapy has gained more attention. The current study demonstrated that epigenetic modification improves the in vitro differentiation of Wharton’s jelly mesenchymal stem cells (WJMSCs) into pancreatic endocrine‐like cells. Here we used two histone deacetylase (HDAC) inhibitors namely trichostatin A (TSA) and TMP269. TSA inhibits both class I and II HDACs whereas TMP269 inhibits only class IIa HDACs. WJMSCs were differentiated using a multistep protocol in a serum‐free condition with or without TSA pretreatment. A marginal improvement in differentiation was observed after TSA pretreatment though it was not significant. However, exposing endocrine precursor‐like cells derived from WJMSCs to TMP269 alone has significantly improved the differentiation toward insulin‐producing cells. Further, increase in the expression of paired box 4 (PAX4), insulin, somatostatin, glucose transporter 2 (GLUT2), MAF bZIP transcription factor A (MAFA), pancreatic duodenal homeobox 1 (PDX‐1), and NKX6.1 was observed both at messenger RNA and protein levels. Nevertheless, TMP269‐treated cells secreted higher insulin upon glucose challenge, and demonstrated increased dithizone staining. These findings suggest that TMP269 may improve the in vitro differentiation of WJMSCs into insulin‐producing cells. - Journal of Cellular Physiology, EarlyView.
  

  October 21, 2018   doi: 10.1002/jcp.27184   open full text
• TRPM4 channel is involved in regulating epithelial to mesenchymal transition, migration, and invasion of prostate cancer cell lines.
  Alfredo I. Sagredo, Eduardo A. Sagredo, Victor Pola, César Echeverría, Rodrigo Andaur, Luis Michea, Andrés Stutzin, Felipe Simon, Katherine Marcelain, Ricardo Armisén.
  Journal of Cellular Physiology. October 21, 2018
  --- - |2- Abstract Transient Receptor Potential Melastatin 4 (TRPM4) is a Ca2+‐activated and voltage‐dependent monovalent cation channel, which depolarizes the plasma cell membrane, thereby modulating Ca2+ influx across Ca2+‐permeable pathways. TRPM4 is involved in different physiological processes such as T cell activation and the migration of endothelial and certain immune cells. Overexpression of this channel has been reported in various types of tumors including prostate cancer. In this study, a significant overexpression of TRPM4 was found only in samples from cancer with a Gleason score higher than 7, which are more likely to spread. To evaluate whether TRPM4 overexpression was related to the spreading capability of tumors, TRPM4 was knockdown by using shRNAs in PC3 prostate cancer cells and the effect on cellular migration and invasion was analyzed. PC3 cells with reduced levels of TRPM4 (shTRPM4) display a decrease of the migration/invasion capability. A reduction in the expression of Snail1, a canonical epithelial to mesenchymal transition (EMT) transcription factor, was also observed. Consistently, these cells showed a significant change in the expression of key EMT markers such as MMP9, E‐cadherin/N‐cadherin, and vimentin, indicating a partial reversion of the EMT process. Whereas, the overexpression of TRPM4 in LnCaP cells resulted in increased levels of Snail1, reduction in the expression of E‐cadherin and increase in their migration potential. This study suggests a new and indirect mechanism of regulation of migration/invasion process by TRPM4 in prostate cancer cells, by inducing the expression of Snail1 gene and consequently, increasing the EMT. - Journal of Cellular Physiology, EarlyView.
  

  October 21, 2018   doi: 10.1002/jcp.27371   open full text
• MiR302c, Sp1, and NFATc2 regulate interleukin‐21 expression in human CD4+CD45RO+ T lymphocytes.
  Hassan El‐Said, Mohammad Fayyad‐Kazan, Rabab Aoun, Nada Borghol, Najwa Skafi, Redouane Rouas, Luc Vanhamme, Mohamad Mourtada, Mohamad Ezzeddine, Arsène Burny, Hussein Fayyad‐Kazan, Bassam Badran.
  Journal of Cellular Physiology. October 21, 2018
  --- - |2- Abstract Interleukin‐21 (IL‐21) is a cytokine with potent regulatory effects on different immune cells. Recently, IL‐21 has been contemplated for use in the treatment of cancers. However, the molecular mechanisms regulating human IL‐21 gene expression has not yet been described. In this study, we initially studied the promoter region and identified the transcription start site. We thereafter described the essential region upstream of the transcription start site and showed the in vivo binding of NFATc2 and SP1 transcription factors to this region, in addition to their positive role in IL‐21 expression. We also studied the role of microRNAs (miRNAs) in regulating IL‐21 expression. We, thus, established the miRNA profile of CD4+CD45RO+ versus CD4+CD45RA+ isolated from healthy volunteers and identified a signature composed of 12 differentially expressed miRNAs. We showed that miR‐302c is able to negatively regulate IL‐21 expression by binding directly to its target site in the 3′‐untranslated region. Moreover, after using fresh human CD4‐positive T cells, we observed the high acetylation level of histone H4, an observation well in line with the already described high expression of IL‐21 in CD4+CD45RO+ versus CD4+CD45RA+ T cells. Altogether, our data identified different molecular mechanisms regulating IL‐21 expression. - Journal of Cellular Physiology, EarlyView.
  

  October 21, 2018   doi: 10.1002/jcp.27151   open full text
• CSC‐3436 inhibits TWIST‐induced epithelial–mesenchymal transition via the suppression of Twist/Bmi1/Akt pathway in head and neck squamous cell carcinoma.
  Ying‐Ju Lai, Wan‐Nien Yu, Sheng‐Chu Kuo, Chi‐Tang Ho, Chao‐Ming Hung, Tzong‐Der Way, Chiung‐Tong Chen.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Head and neck squamous cell carcinoma (HNSCC) is one of the leading causes of cancer deaths worldwide, especially in male. With poor prognosis, significant portions of patients with HNSCC die due to cancer recurrence and tumor metastasis after chemotherapy and targeted therapies. The HNSCC FaDu cell ectopic expression of Twist, a key transcriptional factor of epithelial–mesenchymal transition (EMT), which triggers EMT and results in the acquisition of a mesenchymal phenotype, was used as the cell model. Our results demonstrated that treatment with newly synthesized 2‐(3‐hydroxyphenyl)‐5‐methylnaphthyridin‐4‐one (CSC‐3436), a flavonoid derivative, elicited changes in its cell morphology, upregulated E‐cadherin messenger RNA and protein expression, downregulated N‐cadherin, vimentin, and CD133 (a marker associated with tumor‐initiating cells) in FaDu‐pCDH‐Twist cells. Moreover, CSC‐3436 exposure reduced B cell‐specific Moloney murine leukemia virus integration site 1 (Bmi1) expression regulated by Twist and further suppressed the direct co‐regulation of E‐cadherin by Twist and Bmi1. Interestingly, CSC‐3436 reduced EMT, cancer stemness, and migration/invasion abilities through the inhibition of the Twist/Bmi1‐Akt/β‐catenin pathway. Most importantly, our findings provided new evidence that CSC‐3436 played a crucial role in therapeutic targeting to Bmi1 and its molecular pathway in HNSCC, and it will be valuable in prognostic prediction and treatment. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27589   open full text
• miR‐335‐5p induces insulin resistance and pancreatic islet β‐cell secretion in gestational diabetes mellitus mice through VASH1‐mediated TGF‐β signaling pathway.
  Xu‐Wen Tang, Qing‐Xin Qin.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Multiple studies have reported different methods in treating gestational diabetes mellitus (GDM); however, the relationship between miR‐335‐5p and GDM still remains unclear. Here, this study explores the effect of miR‐335‐5p on insulin resistance and pancreatic islet β‐cell secretion via activation of the TGFβ signaling pathway by downregulating VASH1 expression in GDM mice. The GDM mouse model was established and mainly treated with miR‐335‐5p mimic, miR‐335‐5p inhibitor, si‐VASH1, and miR‐335‐5p inhibitor + si‐VASH1. Oral glucose tolerance test (OGTT) was conducted to detect fasting blood glucose (FBG) fasting insulin (FINS). The OGTT was also used to calculate a homeostasis model assessment of insulin resistance (HOMA‐IR). A hyperglycemic clamp was performed to measure the glucose infusion rate (GIR), which estimated β‐cell function. Expressions of miR‐335‐5p, VASH1, TGF‐β1, and c‐Myc in pancreatic islet β‐cells were determined by RT‐qPCR, western blot analysis, and insulin release by ELISA. The miR‐335‐5p mimic and si‐VASH1 groups showed elevated blood glucose levels, glucose area under the curve (GAUC), and HOMA‐IR, but a reduced GIR and positive expression of VASH1. Overexpression of miR‐335‐5p and inhibition of VASH1 contributed to activated TGFβ1 pathway, higher c‐Myc, and lower VASH1 expressions, in addition to downregulated insulin and insulin release levels. These findings provided evidence that miR‐335‐5p enhanced insulin resistance and suppressed pancreatic islet β‐cell secretion by inhibiting VASH1, eventually activating the TGF‐β pathway in GDM mice, which provides more clinical insight on the GDM treatment. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27406   open full text
• Circular RNA in cardiovascular disease.
  M‐Ashraf Altesha, Tiffany Ni, Afaan Khan, Kexiang Liu, Xiufen Zheng.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Circular RNA (circRNA) are endogenous transcripts that display differential expression across species, developmental stages, and pathologies. Their lack of free ends confers increased stability when compared with linear transcripts, making them ideal candidates for future diagnostic biomarkers and therapeutic interventions. Increasing evidence has implicated circRNA in the pathogenesis of multiple cardiovascular diseases. In this paper, we summarize current understanding of circRNA biogenesis, properties, expression profiles, detection methods, functions, and their implication in cardiac pathologies including/ischemia reperfusion injury, myocardial infarction, cardiac senescence, cardiac fibrosis, cardiomyopathy, cardiac hypertrophy and heart failure, atherosclerosis, coronary artery disease, and aneurysm. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27384   open full text
• Strategies to target energy metabolism in consensus molecular subtype 3 along with Kirsten rat sarcoma viral oncogene homolog mutations for colorectal cancer therapy.
  Gang Wang, Jun‐Jie Wang, Pei‐Hao Yin, Ke Xu, Yu‐Zhu Wang, Feng Shi, Jing Gao, Xing‐Li Fu.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Alterations in cellular energy metabolism play a critical role in colorectal cancer (CRC), which has been identified as the definition of consensus molecular subtypes (CMSs), and CMS3 tumors exhibit energy metabolism signatures along with Kirsten rat sarcoma viral oncogene homolog (KRAS)‐activating mutations. This review summarizes the relationship between CMS3 tumors associated with mutated KRAS and energy metabolism in CRC, especially for the dysregulated energy metabolism that affects tumor cell proliferation, invasion, and migration. Furthermore, this review concentrates on the role of metabolic genes and factors and signaling pathways, which coupled with a primary energy source connected with the CMS3 associated with mutated KRAS, induce metabolic alterations. The strategies to target energy metabolism for the metabolic alterations in mutated KRAS CRC are also introduced. In conclusion, dysregulated energy metabolism has a close relationship with mutated KRAS in CMS3 tumors. Therefore, selective inhibitors or agents against metabolic targets or KRAS signaling may be clinically useful for CMS3 tumor treatment through a personalized approach for patients with cancer. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27388   open full text
• Wnt lipidation: Roles in trafficking, modulation, and function.
  Vahid Hosseini, Christian Dani, Mohammad Hossein Geranmayeh, Fatemeh Mohammadzadeh, Saeed Nazari Soltan Ahmad, Masoud Darabi.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract The Wnt signaling pathway consists of various downstream target proteins that have substantial roles in mammalian cell proliferation, differentiation, and development. Its aberrant activity can lead to uncontrolled proliferation and tumorigenesis. The posttranslational connection of fatty acyl chains to Wnt proteins provides the unique capacity for regulation of Wnt activity. In spite of the past belief that Wnt molecules are subject to dual acylation, it has been shown that these proteins have only one acylation site and undergo monounsaturated fatty acylation. The Wnt monounsaturated fatty acyl chain is more than just a hydrophobic coating and appears to be critical for Wnt signaling, transport, and receptor activation. Here, we provide an overview of recent findings in Wnt monounsaturated fatty acylation and the mechanism by which this lipid moiety regulates Wnt activity from the site of production to its receptor interactions. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27570   open full text
• Behçet’s disease: An immunogenetic perspective.
  Arash Salmaninejad, Mohammad Reza Zamani, Arezoo Gowhari Shabgah, Seyedmojtaba Hosseini, Fatemeh Mollaei, Nayyerehalsadat Hosseini, Amirhossein Sahebkar.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2 Abstract Behçet’s disease (BD) is a chronic and rare multisystemic disorder defined by autoimmunity and inflammatory characteristics, manifested by ocular lesions, recurrent genital and oral ulcers, skin symptoms and arthritis as well as neurological, intestinal, and vascular involvement. Despite the unknown cause of BD, there is some strong documentation for immunological, genetic, environmental, and infectious factors playing a role in the pathogenesis of BD. While the nature of the genetic variants remains unidentified, many genetic risk factors are considered to contribute to BD susceptibility. Along with human leukocyte antigen gene encoding B*51 (HLA‐B*51) and areas including the major histocompatibility complex class I, genome‐wide association studies have recognized numerous other BD susceptibility genes including those encoding interleukin (IL)‐10, IL‐12 receptor β 2 (IL‐12RB2), IL‐23 receptor (IL‐23R), C‐C chemokine receptor 1 gene, signal transducer and activator of transcription 4 (STAT4), endoplasmic reticulum aminopeptidase (ERAP1), and genes encoding killer cell lectin‐like receptor family members (KLRC4‐KLRK1). It is believed that BD could be considered as a disorder lying in between autoimmune and autoinflammatory syndromes. The positive responses to classical immunosuppressive agents like azathioprine and cyclosporine and involvement of autoantigens in the initiation of the disorder are the main BD features that reflect the autoimmune nature of the disorder. In this review, we address recent findings on the role of common cytokines, antibodies and immunogenetic factors in BD. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27576   open full text
• Pancreatic resident endocrine progenitors demonstrate high islet neogenic fidelity and committed homing towards diabetic mice pancreas.
  Abhay Srivastava, Nidheesh Dadheech, Mitul Vakani, Sarita Gupta.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Pancreatic progenitors have been explored for their profound characteristics and unique commitment to generate new functional islets in regenerative medicine. Pancreatic resident endocrine progenitors (PREPs) with mesenchymal stem cell (MSC) phenotype were purified from BALB/c mice pancreas and characterized. PREPs were differentiated into mature islet clusters in vitro by activin‐A and swertisin and functionally characterized. A temporal gene and protein profiling was performed during differentiation. Furthermore, PREPs were labeled with green fluorescent protein (GFP) and transplanted intravenously into streptozotocin (STZ) diabetic mice while monitoring their homing and differentiation leading to amelioration in the diabetic condition. PREPs were positive for unique progenitor markers and transcription factors essential for endocrine pancreatic homeostasis along with having the multipotent MSC phenotype. These cells demonstrated high fidelity for islet neogenesis in minimum time (4 days) to generate mature functional islet clusters (shortest reported period for any isolated stem/progenitor). Furthermore, GFP‐labeled PREPs transplanted in STZ diabetic mice migrated and localized within the injured pancreas without trapping in any other major organ and differentiated rapidly into insulin‐producing cells without an external stimulus. A rapid decrease in fasting blood glucose levels toward normoglycemia along with significant increase in fasting serum insulin levels was observed, which ameliorated the diabetic condition. This study highlights the unique potential of PREPs to generate mature islets within the shortest period and their robust homing toward the damaged pancreas, which ameliorated the diabetic condition suggesting PREPs affinity toward their niche, which can be exploited and extended to other stem cell sources in diabetic therapeutics. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27568   open full text
• Cyclooxygenase‐2 in cancer: A review.
  Nasser Hashemi Goradel, Masoud Najafi, Eniseh Salehi, Bagher Farhood, Keywan Mortezaee.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Cyclooxygenase‐2 (COX‐2) is frequently expressed in many types of cancers exerting a pleiotropic and multifaceted role in genesis or promotion of carcinogenesis and cancer cell resistance to chemo‐ and radiotherapy. COX‐2 is released by cancer‐associated fibroblasts (CAFs), macrophage type 2 (M2) cells, and cancer cells to the tumor microenvironment (TME). COX‐2 induces cancer stem cell (CSC)‐like activity, and promotes apoptotic resistance, proliferation, angiogenesis, inflammation, invasion, and metastasis of cancer cells. COX‐2 mediated hypoxia within the TME along with its positive interactions with YAP1 and antiapoptotic mediators are all in favor of cancer cell resistance to chemotherapeutic drugs. COX‐2 exerts most of the functions through its metabolite prostaglandin E2. In some and limited situations, COX‐2 may act as an antitumor enzyme. Multiple signals are contributed to the functions of COX‐2 on cancer cells or its regulation. Members of mitogen‐activated protein kinase (MAPK) family, epidermal growth factor receptor (EGFR), and nuclear factor‐κβ are main upstream modulators for COX‐2 in cancer cells. COX‐2 also has interactions with a number of hormones within the body. Inhibition of COX‐2 provides a high possibility to exert therapeutic outcomes in cancer. Administration of COX‐2 inhibitors in a preoperative setting could reduce the risk of metastasis in cancer patients. COX‐2 inhibition also sensitizes cancer cells to treatments like radio‐ and chemotherapy. Chemotherapeutic agents adversely induce COX‐2 activity. Therefore, choosing an appropriate chemotherapy drugs along with adjustment of the type and does for COX‐2 inhibitors based on the type of cancer would be an effective adjuvant strategy for targeting cancer. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27411   open full text
• Runx2 is required for postnatal intervertebral disc tissue growth and development.
  Lifan Liao, Hua Jiang, Yunshan Fan, Ronald S. Lu, Changli Wei, Takeshi Takarada, Shisheng He, Di Chen.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Runx2 plays an essential role in embryonic disc tissue development in mice. However, the role of runt‐related transcription factor 2 (Runx2) in postnatal disc tissue growth and development has not been defined. In the present studies, we generated Runx2 conditional knockout (KO) mice (Runx2Agc1ER), in which Runx2 was deleted in Aggrecan‐expressing cells in disc tissue at postnatal 2‐weeks of age. We then analyzed changes in disc tissue growth and development using histology and immunohistochemical methods in 3‐month‐old mice. We found that large vacuolated notochordal cells were accumulated in the nucleus pulposus (NP) in Runx2 KO mice. The growth plate cartilage tissue in the disc was thicker in Runx2 KO mice. We also found a significant upregulation of Indian hedgehog (Ihh) expression in the cells in NP cells and in annulus fibrosus cells of Runx2 KO mice. These results demonstrated that Runx2 may play an important role in postnatal disc tissue development through interacting with Ihh signaling. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27410   open full text
• Homogentisic acid induces morphological and mechanical aberration of ochronotic cartilage in alkaptonuria.
  Giulia Bernardini, Gemma Leone, Lia Millucci, Marco Consumi, Daniela Braconi, Ottavia Spiga, Silvia Galderisi, Barbara Marzocchi, Cecilia Viti, Giovanna Giorgetti, Pietro Lupetti, Agnese Magnani, Annalisa Santucci.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Alkaptonuria (AKU) is a disease caused by a deficient homogentisate 1,2‐dioxygenase activity leading to systemic accumulation of homogentisic acid (HGA), that forms a melanin‐like polymer that progressively deposits onto connective tissues causing a pigmentation called “ochronosis” and tissue degeneration. The effects of AKU and ochronotic pigment on the biomechanical properties of articular cartilage need further investigation. To this aim, AKU cartilage was studied using thermal (thermogravimetry and differential scanning calorimetry) and rheological analysis. We found that AKU cartilage had a doubled mesopore radius compared to healthy cartilage. Since the mesoporous structure is the main responsible for maintaining a correct hydrostatic pressure and tissue homoeostasis, drastic changes of thermal and rheological parameters were found in AKU. In particular, AKU tissue lost its capability to enhance chondrocytes metabolism (decreased heat capacity) and hence the production of proteoglycans. A drastic increase in stiffness and decrease in dissipative and lubricant role ensued in AKU cartilage. Multiphoton and scanning electron microscopies revealed destruction of cell–matrix microstructure and disruption of the superficial layer. Such observations on AKU specimens were confirmed in HGA‐treated healthy cartilage, indicating that HGA is the toxic responsible of morphological and mechanical alterations of cartilage in AKU. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27416   open full text
• Helvolic acid attenuates osteoclast formation and function via suppressing RANKL‐induced NFATc1 activation.
  Kai Chen, Yu Yuan, Ziyi Wang, Dezhi Song, Jinmin Zhao, Zhen Cao, Junhao Chen, Qiang Guo, Li Chen, Jennifer Tickner, Jiake Xu.
  Journal of Cellular Physiology. October 20, 2018
  --- - "\nAbstract\nExcessive osteoclast formation and function are considered as the main causes of bone lytic disorders such as osteoporosis and osteolysis. Therefore, the osteoclast is a potential therapeutic target for the treatment of osteoporosis or other osteoclast‐related diseases. Helvolic acid (HA), a mycotoxin originally isolated from Aspergillus fumigatus , has been discovered as an effective broad‐spectrum antibacterial agent and has a wide range of pharmacological properties. Herein, for the first time, HA was demonstrated to be capable of significantly inhibiting receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclastogenesis and bone resorption in vitro by suppressing nuclear factor of activated T cells 1 (NFATc1) activation. This inhibition was followed by the dramatically decreased expression of NFATc1‐targeted genes including \nCtr (encoding calcitonin receptor), \nAcp5 (encoding tartrate‐resistant acid phosphatase [TRAcP]), \nCtsk (encoding cathepsin K), \nAtp6v0d2 (encoding the vacuolar H+ ATPase V0 subunit d2 [V‐ATPase‐d2]) and \nMmp9 (encoding matrix metallopeptidase 9) which are osteoclastic‐specific genes required for osteoclast formation and function. Mechanistically, HA was shown to greatly attenuate multiple upstream pathways including extracellular signal‐regulated kinase (ERK) phosphorylation, c‐Fos signaling, and intracellular Ca\n2+ oscillation, but had little effect on nuclear factor‐κB (NF‐κB) activation. In addition, HA also diminished the RANKL‐induced generation of intracellular reactive oxygen species. Taken together, our study indicated HA effectively suppressed RANKL‐induced osteoclast formation and function. Thus, we propose that HA can be potentially used in the development of a novel drug for osteoclast‐related bone diseases." - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27385   open full text
• The role of sphingosine 1 phosphate in coronary artery disease and ischemia reperfusion injury.
  Aynaz Mihanfar, Hamid Reza Nejabati, Amir Fattahi, Zeinab Latifi, Masoud Pezeshkian, Abbas Afrasiabi, Naser Safaie, Ahmad Reza Jodati, Mohammad Nouri.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Coronary artery disease (CAD) is a common cause of morbidity and mortality worldwide. Atherosclerotic plaques, as a hallmark of CAD, cause chronic narrowing of coronary arteries over time and could also result in acute myocardial infarction (AMI). The standard treatments for ameliorating AMI are reperfusion strategies, which paradoxically result in ischemic reperfusion (I/R) injury. Sphingosine 1 phosphate (S1P), as a potent lysophospholipid, plays an important role in various organs, including immune and cardiovascular systems. In addition, high‐density lipoprotein, as a negative predictor of atherosclerosis and CAD, is a major carrier of S1P in blood circulation. S1P mediates its effects through binding to specific G protein‐coupled receptors, and its signaling contributes to a variety of responses, including cardiac inflammation, dysfunction, and I/R injury protection. In this review, we will focus on the role of S1P in CAD and I/R injury as a potential therapeutic target. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27353   open full text
• Signatures of altered long noncoding RNAs and messenger RNAs expression in the early acute phase of spinal cord injury.
  Zhongju Shi, Guangzhi Ning, Bin Zhang, Shiyang Yuan, Hengxing Zhou, Bin Pan, Jiahe Li, Zhijian Wei, Fujiang Cao, Xiaohong Kong, Shiqing Feng.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Spinal cord injury (SCI) is a highly severe disease and it can lead to the destruction of the motor and sensory function resulting in temporary or permanent disability. Long noncoding RNAs (lncRNAs) are transcripts longer than 200 nt that play a critical role in central nervous system (CNS) injury. However, the exact roles of lncRNAs and messenger RNAs (mRNAs) in the early acute phase of SCI remain to be elucidated. We examined the expression of mRNAs and lncRNAs in a rat model at 2 days after SCI and identified the differentially expressed lncRNAs (DE lncRNAs) and differentially expressed mRNAs (DE mRNAs) using microarray analysis. Subsequently, a comprehensive bioinformatics analysis was also performed to clarify the interaction between DE mRNAs. A total of 3,193 DE lncRNAs and 4,308 DE mRNAs were identified between the injured group and control group. Classification, length distribution, and chromosomal distribution of the dysregulated lncRNAs were also performed. The gene ontology analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were performed to identify the critical biological processes and pathways. A protein−protein interaction (PPI) network indicated that IL6, TOP2A, CDK1, POLE, CCNB1, TNF, CCNA2, CDC20, ITGAM, and MYC were the top 10 core genes. The subnetworks from the PPI network were identified to further elucidate the most significant functional modules of the DE mRNAs. These data may provide novel insights into the molecular mechanism of the early acute phase of SCI. The identification of lncRNAs and mRNAs may offer potential diagnostic and therapeutic targets for SCI. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27560   open full text
• Circ‐U2AF1 promotes human glioma via derepressing neuro‐oncological ventral antigen 2 by sponging hsa‐miR‐7‐5p.
  Guoxiong Li, Min Huang, Yingqian Cai, Yuantao Yang, Xinlin Sun, Yiquan Ke.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract The prognosis for human glioma, a malignant tumor of the central nervous system, is poor due to its rapid growth, genetic heterogeneity, and inadequate understanding of its underlying molecular mechanisms. Circular RNAs composed of exonic sequences, represent an understudied form of noncoding RNAs (ncRNAs) that was discovered more than a decade ago, function as microRNA sponges. We aimed to assess the relationship between circ‐U2AF1 (CircRNA ID: hsa_circ_0061868) and hsa‐mir‐7‐5p and examine their effects on proliferation, apoptosis, and the metastatic phenotype of glioma cells regulated by neuro‐oncological ventral antigen 2 (NOVA2). We found that the expression levels of circ‐U2AF1 and NOVA2 were upregulated, while hsa‐miR‐7‐5p was downregulated in human glioma tissues and glioma cell lines. Our data and bioinformatic analysis indicated the association of these molecules with glioma grade, a positive correlation between circ‐U2AF1 and NOVA2 expression levels and a negative correlation of hsa‐miR‐7‐5p with both circ‐U2AF1 and NOVA2, respectively. In addition, silencing of circ‐U2AF1 expression resulted in increased hsa‐miR‐7‐5p expression and decreased NOVA2 expression both in vitro and in vivo. Luciferase assay confirmed hsa‐miR‐7‐5p as a direct target of circ‐U2AF1 and NOVA2 as a direct target of hsa‐miR‐7‐5p. Functionally, silencing of circ‐U2AF1 inhibits glioma development by repressing NOVA2 via upregulating hsa‐miR‐7‐5p both in vitro and in vivo. Thus, we assumed that circ‐U2AF1 promotes glioma malignancy via derepressing NOVA2 by sponging hsa‐miR‐7‐5p. Taken together, we suggest that circ‐U2AF1 can be a prognostic biomarker and the circ‐U2AF1/hsa‐miR‐7‐5p/NOVA2 regulatory pathway may be a novel therapeutic target for treating gliomas. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27591   open full text
• Upregulation of long noncoding RNA XIST is associated with poor prognosis in human cancers.
  Ji‐Long Liu, Wen‐Qian Zhang, Miao Zhao, Ming‐Yu Huang.
  Journal of Cellular Physiology. October 20, 2018
  --- - "\nAbstract\nGrowing evidence from recent studies has shown that the X‐inactive specific transcript (XIST), a well‐known long noncoding RNA involved in early embryonic development, is aberrantly regulated in various human cancers. However, the prognostic value of XIST in cancers remains uncharacterized. In this study, we searched PubMed, Web of Science, and Embase to collect all relevant studies, and a meta‐analysis was performed to explore the association of XIST expression with overall survival (OS) and clinicopathological parameters. We demonstrated that high XIST expression was associated with poor OS (hazard ratio = 1.76; 95% confidence intervals [CI], 1.56–1.98; p < 0.001). In addition, increased XIST expression was found to be associated with lymph node metastasis (odds ratio [OR] = 2.06; 95% CI, 1.46–1.90; \np < 0.001), distant metastasis (OR = 2.93; 95% CI, 2.00–4.28; \np < 0.001), tumor size (OR = 2.66; 95% CI, 1.86–3.81; \np < 0.001), poor differentiation (OR = 1.45; 95% CI, 1.00–2.10; \np = 0.049), and advanced tumor stage (OR = 3.35; 95% CI, 2.25–5.00; \np < 0.001), but not with age (OR = 0.82; 95% CI, 0.59–1.15; \np = 0.251) or gender (OR = 0.92; 95% CI, 0.70–1.19; \np = 0.512). Our meta‐analysis showed that XIST may be a useful common biomarker for predicting prognosis in patients with cancer." - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27400   open full text
• Prenatal exposure to valproate induces sex‐, age‐, and tissue‐dependent alterations of cholesterol metabolism: Potential implications on autism.
  Veronica Cartocci, Claudia Tonini, Tiziana Di Pippo, Florenzia Vuono, Sara Schiavi, Maria Marino, Viviana Trezza, Valentina Pallottini.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Here, we investigated the protein network regulating cholesterol metabolism in the liver and brain of adolescent and adult male and female rats prenatally exposed to valproate (VPA), a well validated experimental model of autism spectrum disorders (ASD). We were aimed at studying whether prenatal VPA exposure affected the proteins involved in cholesterol homeostasis in a sex‐dependent manner. To this aim the protein network of cholesterol metabolism, in term of synthesis and plasma membrane trafficking, was analyzed by western blot in the liver and different brain areas (amygdala, cerebellum, cortex, hippocampus, nucleus accumbens, and dorsal striatum) of adolescent and adult male and female rats prenatally exposed to VPA. Our results show that physiological sex‐dependent differences are present both in the liver and in brain of rats. Interestingly, VPA affects specifically the brain in an age‐ and region‐specific manner; indeed, cerebellum, cortex, hippocampus and nucleus accumbens are affected in a sex‐dependent way, while this does not occur in amygdala and dorsal striatum. Overall, we demonstrate that each brain area responds differently to the same external stimulus and males and females respond in a different way, suggesting that this could be related to the diverse incidences, between the sexes, of some neurodevelopmental pathologies such as autism, which displays a 3:1 male to female ratio. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27218   open full text
• Heat stress induces distinct responses in porcine cumulus cells and oocytes associated with disrupted gap junction and trans‐zonal projection colocalization.
  Chao Yin, Jie Liu, Bin He, Longfei Jia, Yabin Gong, Huiduo Guo, Ruqian Zhao.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Cumulus cells (CCs), the granulosa cells surrounding the oocytes, play critical roles in oocytes maturation through intercellular communication by extending trans‐zonal projections (TZPs) to contact oocytes via gap junctions (GJs). The adverse effect of heat stress (HS) on oocyte maturation has been well documented, whereas the HS responses of CCs and the oocytes in association with GJ/TZP colocalization remain unclear. In this study, porcine cumulus‐oocyte complexes (COCs) were subjected to HS at 41.5°C for 24 hr during in vitro maturation. Cumulus expansion was impaired and oocyte quality was reduced with lower survival rate, polar body extrusion rate, and early embryo developmental potentials. CCs and oocytes isolated from COCs demonstrated distinct responses to HS. The messenger RNA abundance of heat shock protein‐related genes and mitochondrial DNA‐encoded genes, together with ATP content, were significantly increased in CCs, yet decreased in oocytes, despite activation of caspase 3 detected in both CCs and oocytes. Similar changes were observed when denuded oocytes and isolated CCs subjected to HS separately, except mitochondria reactive oxygen species (mROS). In heat‐stressed COCs, mROS was significantly increased only in oocytes. However, when isolated CCs and denuded oocytes were heat‐stressed separately, mROS was significantly increased only in CCs. Moreover, F‐actin, a TZP marker, and its colocalization with a GJ protein connexin‐45, were significantly reduced in heat‐exposed COCs. These results indicate that HS induces distinct responses in porcine CCs and oocytes in association with disrupted GJ and TZP colocalization. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27277   open full text
• Colorectal cancer invasiveness in vitro: Predominant contribution of neonatal Nav1.5 under normoxia and hypoxia.
  R. Mine Guzel, Kazim Ogmen, Kristina M. Ilieva, Scott P. Fraser, Mustafa B. A. Djamgoz.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract Functional expression of voltage‐gated Na+ channels (VGSCs) occurs in human carcinomas and promotes invasiveness in vitro and metastasis in vivo. Both neonatal and adult forms of Nav1.5 (nNav1.5 and aNav1.5, respectively) have been reported to be expressed at messenger RNA (mRNA) level in colorectal cancer (CRCa) cells. Here, three CRCa cell lines (HT29, HCT116 and SW620) were studied and found to express nNav1.5 mRNA and protein. In SW620 cells, adopted as a model, effects of gene silencing (by several small interfering RNAs [siRNAs]) selectively targeting nNav1.5 or aNav1.5 were determined on (a) channel activity and (b) invasiveness in vitro. Silencing nNav1.5 made the currents more “adult‐like” and suppressed invasion by up to 73%. Importantly, subsequent application of the highly specific, general VGSC blocker, tetrodotoxin (TTX), had no further effect. Conversely, silencing aNav1.5 made the currents more “neonatal‐like” but suppressed invasion by only 17% and TTX still induced a significant effect. Hypoxia increased invasiveness and this was also blocked completely by siRNA targeting nNav1.5. The effect of hypoxia was suppressed dose dependently by ranolazine, but its effect was lost in cells pretreated with nNav1.5‐siRNA. We conclude that (a) functional nNav1.5 expression is common to human CRCa cells, (b) hypoxia increases the invasiveness of SW620 cells, (c) the VGSC‐dependent invasiveness is driven predominantly by nNav1.5 under both normoxic and hypoxic conditions and (d) the hypoxia‐induced increase in invasiveness is likely to be mediated by the persistent current component of nNav1.5. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27399   open full text
• Necrosis and necroptosis in germ cell depletion from mammalian ovary.
  Govind R. Chaudhary, Pramod K. Yadav, Anil K. Yadav, Meenakshi Tiwari, Anumegha Gupta, Alka Sharma, Kankshi Sahu, Ashutosh N. Pandey, Ajai K. Pandey, Shail K. Chaube.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract The maximum number of germ cells is present during the fetal life in mammals. Follicular atresia results in rapid depletion of germ cells from the cohort of the ovary. At the time of puberty, only a few hundred (<1%) germ cells are either culminated into oocytes or further get eliminated during the reproductive life. Although apoptosis plays a major role, necrosis as well as necroptosis, might also be involved in germ cell elimination from the mammalian ovary. Both necrosis and necroptosis show similar morphological features and are characterized by an increase in cell volume, cell membrane permeabilization, and rupture that lead to cellular demise. Necroptosis is initiated by tumor necrosis factor and operated through receptor interacting protein kinase as well as mixed lineage kinase domain‐like protein. The acetylcholinesterase, cytokines, starvation, and oxidative stress play important roles in necroptosis‐mediated granulosa cell death. The granulosa cell necroptosis directly or indirectly induces susceptibility toward necroptotic or apoptotic cell death in oocytes. Indeed, prevention of necrosis and necroptosis pathways using their specific inhibitors could enhance growth/differentiation factor‐9 expression, improve survivability as well as the meiotic competency of oocytes, and prevent decline of reproductive potential in several mammalian species and early onset of menopause in women. This study updates the information and focuses on the possible involvement of necrosis and necroptosis in germ cell depletion from the mammalian ovary. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27562   open full text
• Downregulation of SPIN90 promotes fibroblast activation via periostin‐FAK‐ROCK signaling module.
  Eunae You, Yun‐Hyun Huh, Jieun Lee, Panseon Ko, Jangho Jeong, Seula Keum, Jaegu Kim, Ahreum Kwon, Woo Keun Song, Sangmyung Rhee.
  Journal of Cellular Physiology. October 20, 2018
  --- - "\nAbstract\nAlterations in mechanical properties in the extracellular matrix are modulated by myofibroblasts and are required for progressive fibrotic diseases. Recently, we reported that fibroblasts depleted of SPIN90 showed enhanced differentiation into myofibroblasts via increased acetylation of microtubules in the soft matrix; the mechanisms of the underlying signaling network, however, remain unclear. In this study, we determine the effect of depletion of SPIN90 on FAK/ROCK signaling modules. Transcriptome analysis of \nSpin90 KO mouse embryonic fibroblasts (MEF) and fibroblasts activated by TGF‐β revealed that \nPostn is the most significantly upregulated gene. Knockdown of \nPostn by small interfering RNA suppressed cell adhesion and myofibroblastic differentiation and downregulated FAK activity in \nSpin90 KO MEF. Our results indicate that SPIN90 depletion activates FAK/ROCK signaling, induced by \nPostn expression, which is critical for myofibroblastic differentiation on soft matrices mimicking the mechanical environment of a normal tissue." - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27600   open full text
• Long noncoding RNA OPA‐interacting protein 5 antisense transcript 1 upregulated SMAD3 expression to contribute to metastasis of cervical cancer by sponging miR‐143‐3p.
  Xing Chen, Dongsheng Xiong, Huichun Yang, Liya Ye, Shuangshuang Mei, Jinhong Wu, Shanshan Chen, Xianwen Shang, Kai Wang, Lingfei Huang.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2 Abstract Objectives SMAD3 is pivotal in the biology functions of various tumors. This study is aiming to study the relationship among SMAD3, long noncoding RNAs (lncRNAs) OPA‐interacting protein 5 antisense transcript 1 (OIP5‐AS1), and miR‐143‐3p, and their effects on cervical cancer. Methods In our research, real‐time polymerase chain reaction and western blot assay were conducted to detect the expression level of messenger RNA and protein in tumor tissues and cells. Transfection of lncRNA OIP5‐AS1, miR‐143‐3p, or SMAD3 was performed to investigate their potential effects on the function of cell as well as the relationship among them in cervical cell lines via 3‐(4,5‐dimethylthiazolyl‐2)‐2,5‐diphenyltetrazolium bromide) together with transwell assays or dual‐luciferase reporter assay respectively. Results SMAD3, lncRNA OIP5‐AS1 expression is significantly enhanced in cervical cancer tissues and cell lines, but miR‐143‐3p was inhibited. LncRNA OIP5‐AS1 is demonstrated to mediate the physiological process of cervical cancer cells. Moreover, silencing SMAD3 via siRNA suppressed cell number, viability, migration and invasion, whereas overexpression of OIP5‐AS1 promoted these abilities. Furthermore, lncRNA OIP5‐AS1 exert its function via sponging miR‐143‐3p to regulate SMAD3 expression. Conclusions LncRNA OIP5‐AS1 promoted SMAD3 expression via mediating miR‐143‐3p to promote migration and invasion of cervical cancer cells. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27336   open full text
• YAP regulates periodontal ligament cell differentiation into myofibroblast interacted with RhoA/ROCK pathway.
  Yao He, Hui Xu, Zichao Xiang, Hongyou Yu, Li Xu, Yongwen Guo, Ye Tian, Rui Shu, Xianrui Yang, Chaoran Xue, Mengyuan Zhao, Yiruo He, Xianglong Han, Ding Bai.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2- Abstract During orthodontic tooth movement (OTM), periodontal ligament cells (PDLCs) receive the mechanical stimuli and transform it into myofibroblasts (Mfbs). Indeed, previous studies have demonstrated that mechanical stimuli can promote the expression of Mfb marker α‐smooth muscle actin (α‐SMA) in PDLCs. Transforming growth factor β1 (TGF‐β1), as the target gene of yes‐associated protein (YAP), has been proven to be involved in this process. Here, we sought to assess the role of YAP in Mfbs differentiation from PDLCs. The time‐course expression of YAP and α‐SMA was manifested in OTM model in vivo as well as under tensional stimuli in vitro. Inhibition of RhoA/Rho‐associated kinase (ROCK) pathway using Y27632 significantly reduced tension‐induced Mfb differentiation and YAP expression. Moreover, overexpression of YAP with lentiviral transfection in PDLCs rescued the repression effect of Mfb differentiation induced by Y27632. These data together suggest a crucial role of YAP in regulating tension‐induced Mfb differentiation from PDLC interacted with RhoA/ROCK pathway. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27312   open full text
• MiR‐128‐3p accelerates cardiovascular calcification and insulin resistance through ISL1‐dependent Wnt pathway in type 2 diabetes mellitus rats.
  Xin‐Yong Wang, Xian‐Zhao Zhang, Feng Li, Qing‐Rong Ji.
  Journal of Cellular Physiology. October 20, 2018
  --- - |2 Abstract Vascular calcification is highly prevalent in patients with type 2 diabetes mellitus (T2DM), one of the most common chronic diseases with high morbidity and mortality. In recent years, microRNAs have been widely reported as potential biomarkers for the diagnosis and treatment of T2DM. We hypothesized that miR‐128‐3p is associated with cardiovascular calcification and insulin resistance (IR) in rats with T2DM by targeting ISL1 via the Wnt pathway. Microarray analysis was adopted to identify differentially expressed genes related to T2DM. T2DM models were induced in rats. Blood samples from normal and T2DM rats were used to detect islet β‐cell function, islet sensitivity, and calcium content. Next, islet tissues were obtained to identify the expression of miR‐128‐3p, ISL1, and the Wnt signaling pathway‐ and apoptosis‐related genes. Finally, apoptosis of islet β‐cells was determined by flow cytometry. Through microarray analysis of GSE27382 and GSE23343, ISL1 was found to be downregulated in T2DM. In blood samples from T2DM rats, basic biochemical indicators, IR, and calcium content were increased, and islet sensitivity and islet β‐cell function were decreased. Furthermore, upregulation of miR‐128‐3p and ISL1 gene silencing promoted the expression of Wnt‐1, β‐catenin, GSK‐3β, and Bax and the phosphorylation of β‐catenin and GSK‐3β, inhibited c‐fos, PDX‐1, and Bcl‐2 expression, and enhanced cell apoptosis. The key findings of our study demonstrate that miR‐128‐3p aggravates cardiovascular calcification and IR in T2DM rats by downregulating ISL1 through the activation of the Wnt pathway. Thus, miR‐128‐3p may serve as a potential target for the treatment of T2DM. - Journal of Cellular Physiology, EarlyView.
  

  October 20, 2018   doi: 10.1002/jcp.27300   open full text
• Combination therapy for the treatment of pancreatic cancer through hyaluronic acid‐decorated nanoparticles loaded with quercetin and gemcitabine: A preliminary in vitro study.
  Carla Serri, Vincenzo Quagliariello, Rosario Vincenzo Iaffaioli, Sabato Fusco, Gerardo Botti, Laura Mayol, Marco Biondi.
  Journal of Cellular Physiology. October 18, 2018
  --- - |2- Abstract Combination chemotherapy by means of two or more drugs is prone to suppressing or discouraging the inception of multidrug resistance, exploiting the fact that diverse drugs act in different points of the cellular cycle of amplifying tumor cells. For example, the combination of gemcitabine (GMC) with quercetin (QCT) showed a synergistic effect in inhibiting the migration of pancreatic cancer cells. Consequently, herein GMC and QCT have been loaded within biodegradable nanoparticles (NPs) based on poly(lactic‐co‐glycolic acid), externally decorated with hyaluronic acid (HA; viz., PPHA NPs), which plays a major role in drug targeting to tumors due to its ability to specifically interact with CD44 receptor, that is overexpressed in many tumors. The produced HA‐decorated NPs loaded with GMC and QCT showed an improved cytotoxicity and cellular uptake toward two cell lines of pancreatic ductal adenocarcinoma, namely Mia‐PaCa‐2 and PANC‐1, compared with both the bare drugs and the drugs loaded in NPs which do not expose HA on the surface. HA‐decorated NPs were also able to improve the anti‐inflammatory properties of QCT, therefore leading to a decrease of interleukin cellular levels in both cell lines, preliminarily stimulated with lipopolysaccharides. This result is of special interest also considering the crucial role of interleukins in progression, metastatic processes, and drug resistance of human pancreas cancer cells. - Journal of Cellular Physiology, EarlyView.
  

  October 18, 2018   doi: 10.1002/jcp.27297   open full text
• Metformin inhibits mTOR–HIF‐1α axis and profibrogenic and inflammatory biomarkers in thioacetamide‐induced hepatic tissue alterations.
  Fahaid Al‐Hashem, Suliman Al‐Humayed, Shaimaa N. Amin, Samaa S. Kamar, Soheir S. Mansy, Sarah Hassan, Lubna O. Abdel‐Salam, Mohamed Abd Ellatif, Mohammed Alfaifi, Mohamed A. Haidara, Bahjat Al‐Ani.
  Journal of Cellular Physiology. October 18, 2018
  --- - |2- Abstract The potential inhibitory effect of the antidiabetic and anti‐inflammatory drug, metformin on thioacetamide (TAA)‐induced hepatotoxicity associated with the inhibition of mammalian target of rapamycin (mTOR)–hypoxia‐inducible factor‐1α (HIF‐1α) axis has not been investigated before. Therefore, we tested whether metformin can protect against liver injuries including fibrosis induced by TAA possibly via the downregulation of mTOR–HIF‐1α axis and profibrogenic and inflammatory biomarkers. Rats either injected with TAA (200 mg/kg; twice a week for 8 weeks) before being killed after 10 weeks (model group) or were pretreated with metformin (200 mg/kg) daily for 2 weeks before TAA injections and continued receiving both agents until the end of the experiment, at Week 10 (protective group). Using light and electron microscopy examinations, we observed in the model group substantial damage to the hepatocytes and liver tissue such as collagen deposition, infiltration of inflammatory cells, and degenerative cellular changes with ballooned mitochondria that were substantially ameliorated by metformin. Metformin also significantly ( p < 0.05) inhibited TAA‐induced HIF‐1α, mTOR, the profibrogenic biomarker α‐smooth muscle actin, tissue inhibitor of metalloproteinases‐1, tumor necrosis factor‐α (TNF‐α), interleukin‐6 (IL‐6), alanine aminotransferase (ALT) and aspartate aminotransferase in harvested liver homogenates and blood samples. In addition, a significant ( p < 0.01) positive correlation between hypoxia scoring (HIF‐1α) and the serum levels of TNF‐α ( r = 0.797), IL‐6 ( r = 0.859), and ALT ( r = 0.760) was observed. We conclude that metformin protects against TAA‐induced hepatic injuries in rats, which is associated with the inhibition of mTOR–HIF‐1α axis and profibrogenic and inflammatory biomarkers; thus, may offer therapeutic potential in humans. - Journal of Cellular Physiology, EarlyView.
  

  October 18, 2018   doi: 10.1002/jcp.27616   open full text
• Characterizing the three‐dimensional organization of telomeres in papillary thyroid carcinoma cells.
  Paola Caria, Tinuccia Dettori, Daniela Virginia Frau, Daniel Lichtenzstejn, Fabiana Pani, Roberta Vanni, Sabine Mai.
  Journal of Cellular Physiology. October 17, 2018
  --- - |2- Abstract The relationship between the three‐dimensional (3D) nuclear telomere architecture and specific genetic alterations in papillary thyroid carcinoma (PTC), in particular in cancer stem‐like cells (CSLCs), has not yet been investigated. We isolated thyrospheres containing CSLCs from B‐CPAP, K1, and TPC‐1 PTC‐derived cell lines, representative of tumors with different genetic backgrounds within the newly identified BRAFV600E‐like PTC subgroup, and used immortalized normal human thyrocytes (Nthy‐ori 3.1) as control. We performed quantitative fluorescence in situ hybridization, 3D imaging, and 3D telomere analysis using TeloView software to examine telomere dysfunction in both parental and thyrosphere cells. Among the 3D telomere profile, a wide heterogeneity was observed, except for telomere intensity. Our findings indicate that CSLCs of each cell line had longer telomeres than parental cells, according to telomere intensity values, which correlate with telomere length. Indeed, the thyrosphere cells had lower numbers of lower‐intensity telomeres (≤5,000 arbitrary fluorescent units, a.u.), compared with parental cancer cells, as well as parental control cells, (p < 0.0001). The B‐CPAP thyrospheres showed a decreased number of higher intensity telomeres (>17,000 a.u.) than K1 and TPC‐1 cells, as well as control cells (p < 0.0001). By selecting PTC‐derived cell lines with different genetic backgrounds characteristic of BRAFV600E‐like PTC subgroups, we demonstrate that thyrosphere cells with BRAFV600E and TP53 mutations show shorter telomeres than those harboring RET/PTC or BRAFV600Eand wild‐type TP53. Hence, our data reveal a trend towards a decrease in telomere shortening in CSLCs, representing the early cancer‐promoting subpopulation, as opposed to parental cells representing the tumor bulk cells. - Journal of Cellular Physiology, EarlyView.
  

  October 17, 2018   doi: 10.1002/jcp.27321   open full text
• The competing endogenous circular RNA ADAMTS14 suppressed hepatocellular carcinoma progression through regulating microRNA‐572/regulator of calcineurin 1.
  Changlong Song, Dianqiu Li, Hongyu Liu, Hongyan Sun, Zhen Liu, Lirong Zhang, Yu Hu.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nEmerging evidence have discovered that circular RNAs (circRNAs) may serve as diagnostic or tumor promising biomarkers. This study aimed to investigate how circular RNA ADAMTS14 (circADAMTS14) regulates microRNA‐572/\nregulator of calcineurin 1(miR‐572/\nRCAN1) in hepatocellular carcinoma (HCC). The expression profiles of circRNA/microRNA (mRNA) between HCC tissues and paired adjacent tissues were analyzed via microarray analysis. The expressions of circADAMTS14, miR‐572, and \nRCAN1 were measured by real‐time polymerase chain reaction (PCR). The protein expression level of \nRCAN1 in HCC cells was detected by western blot. The viability and apoptosis levels of HCC cell lines were measured by the cell counting Kit‐8 (CCK‐8) assay and fluorescence‐activated cell sorter. The invasiveness and migration of cells were detected based on the transwell and wound‐healing assay, respectively. The dual‐luciferase reporter assays were used to reveal circADAMTS14 and \nRCAN1 as a potential target of miR‐572, which was predicted by TargetScan and miRBase. The effect of circADAMTS14 on HCC cells was demonstrated by tumor formation in nude mice in vivo. CircADAMTS14 and \nRCAN1 were lowly expressed in HCC clinical specimens and cell lines using microarrays and qRT‐PCR, but miR‐572 inversely. Our study further verified the direct interaction between circADAMTS14 and \nRCAN1 with miR‐572 via the dual‐luciferase reporter gene assay. Overexpressed circADAMTS14 and \nRCAN1 induced apoptosis of HCC cells and inhibited cell proliferation and invasion. But overexpressed miR‐572 could decrease apoptosis of HCC cells and promote proliferation and invasion. In vivo, circADAMTS14 inhibited the tumor growth, correlated positively with the protein expression levels of \nRCAN1. Our results demonstrated that circADAMTS14 might suppress HCC progression through regulating miR‐572/\nRCAN1 as the competing endogenous RNA." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.26764   open full text
• Lipopolysaccharide‐induced proliferation and glycolysis in airway smooth muscle cells via activation of Drp1.
  Lixin Zhang, Cui Ma, Xiaoying Wang, Siyu He, Qian Li, Yutian Zhou, Ying Liu, Min Zhang, Xiufeng Yu, Xijuan Zhao, Fei Li, Da‐ling Zhu.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Abnormal airway smooth muscle cells (ASMCs) proliferation is an important pathological process in airway remodeling contributes to increased mortality in asthma. Mitochondrial dynamics and metabolism have a central role in the maintenance of the cell function. In this study, lipopolysaccharide (LPS)‐induced ASMCs proliferative model was used to investigate the effect of mitochondria on the proliferation of ASMCs and the possible mechanism. We used cell and molecular biology to determine the effect of dynamin‐related protein 1 (Drp1) on LPS‐mediated ASMCs cell cycle progression and glycolysis. The major findings of the current study are as follows: LPS promoted an increased mitochondrial fission and phosphorylation of Drp1 at Ser616 (p‐Drp1 Ser616). LPS‐induced ASMCs proliferation and cell cycle progression, which was significantly inhibited application of Drp1 RNA interfering. Glycolysis inhibitor 2‐deoxyglucose (2‐DG) depressed ASMCs proliferative process induced by LPS stimulation. LPS caused mitochondrial metabolism disorders and aerobic glycolysis in a dependent on Drp1 activation. These results indicated that Drp1 may function as a key factor in asthma airway remodeling by mediating ASMC proliferation and cell cycle acceleration through an effect on mitochondrial metabolic disturbance. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27605   open full text
• Genome‐wide CRISPR‐Cas9 viability screen reveals genes involved in TNF‐α‐induced apoptosis of human umbilical vein endothelial cells.
  Meng Cai, Sitao Li, Yunfei Shuai, Jie Li, Jieqiong Tan, Qiyi Zeng.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nTumor necrosis factor α (TNF‐α), a pivotal cytokine in sepsis, protects the host against pathogens by promoting an inflammatory response while simultaneously inducing apoptosis of the vascular endothelium. Unfortunately, inhibitors targeting certain components of the TNF‐α signaling pathway to reduce cellular apoptosis have failed to translate into clinical applications, partly due to the adverse effects of excessive immunosuppression. In an attempt to discover potential targets in the TNF‐α signaling pathway to modulate moderate inflammation and apoptosis during the development of sepsis, we performed a pooled genome‐wide CRISPR/Cas9 knockout screen in human umbilical vein endothelial cells (HUVECs). Tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), B‐cell lymphoma 2 (BCL2), Bcl2‐associated death promoter (BAD), and NLR family member X1 (NLRX1) deficiencies were identified as the effective genetic suppressors of TNF‐α cytotoxicity on a list of candidate regulators. CRISPR‐mediated \nNLRX1 knockout conferred cellular resistance to challenge with TNF‐α, and NLRX1 could be induced to colocalize with mitochondria following TNF‐α stimulation. Thus, our work demonstrates the advantage of genome‐scale screening with Cas9 and validates NLRX1 as a potential modulator of TNF‐α‐induced vascular endothelial apoptosis during sepsis." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27595   open full text
• Identification of TAF1, HNF4A, and CALM2 as potential therapeutic target genes for liver fibrosis.
  Dong Ji, Guo‐Feng Chen, Jin‐Cheng Wang, Li‐Hua Cao, Fengmin Lu, Xiao‐Xin Mu, Xiao‐Yu Zhang, Xiao‐Jie Lu.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract The molecular mechanism of liver fibrosis caused by hepatitis C virus (HCV) is not clear. The aim of this study is to understand the molecular mechanism of liver fibrosis induced by HCV and to identify potential therapeutic targets for hepatic fibrosis. We analyzed gene expression patterns between high liver fibrosis and low liver fibrosis samples, and identified genes related to liver fibrosis. We identified TAF1, HNF4A, and CALM2 were related to the development of liver fibrosis. HNF4A is important for hepatic fibrogenesis, and upregulation of HNF4A is an ideal choice for treating liver fibrosis. The gene expression of CALM2 is significantly lower in liver fibrosis samples than nonfibrotic samples. TAF1 may serve as a biomarker for liver fibrosis. The results were further validated by an independent data set GSE84044. In summary, our study described changes in the gene expression during the occurrence and development of liver fibrosis. The TAF1, HNF4A, and CALM2 may serve as novel targets for the treatment of liver fibrosis. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27579   open full text
• A pH‐sensitive luminal His‐cluster promotes interaction of PAM with V‐ATPase along the secretory and endocytic pathways of peptidergic cells.
  Vishwanatha K. Rao, Gerardo Zavala, Abhijit Deb Roy, Richard E. Mains, Betty A. Eipper.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract The biosynthetic and endocytic pathways of secretory cells are characterized by progressive luminal acidification, a process which is crucial for posttranslational modifications and membrane trafficking. This progressive fall in luminal pH is mainly achieved by the vacuolar‐type‐H+ ATPase (V‐ATPase). V‐ATPases are large, evolutionarily ancient rotary proton pumps that consist of a peripheral V1 complex, which hydrolyzes ATP, and an integral membrane V0 complex, which transports protons from the cytosol into the lumen. Upon sensing the desired luminal pH, V‐ATPase activity is regulated by reversible dissociation of the complex into its V1 and V0 components. Molecular details of how intraluminal pH is sensed and transmitted to the cytosol are not fully understood. Peptidylglycine α‐amidating mono‐oxygenase (PAM; EC 1.14.17.3), a secretory pathway membrane enzyme which shares similar topology with two V‐ATPase accessory proteins (Ac45 and prorenin receptor), has a pH‐sensitive luminal linker region. Immunofluorescence and sucrose gradient analysis of peptidergic cells (AtT‐20) identified distinct subcellular compartments exhibiting spatial co‐occurrence of PAM and V‐ATPase. In vitro binding assays demonstrated direct binding of the cytosolic domain of PAM to V1H. Blue native PAGE identified heterogeneous high‐molecular weight complexes of PAM and V‐ATPase. A PAM‐1 mutant (PAM‐1/H3A) with altered pH sensitivity had diminished ability to form high‐molecular weight complexes. In addition, V‐ATPase assembly status was altered in PAM‐1/H3A expressing cells. Our analysis of the secretory and endocytic pathways of peptidergic cells supports the hypothesis that PAM serves as a luminal pH‐sensor, regulating V‐ATPase action by altering its assembly status. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27528   open full text
• Ovarian cancer stem cell: A potential therapeutic target for overcoming multidrug resistance.
  Aynaz Mihanfar, Javad Aghazadeh Attari, Iraj Mohebbi, Maryam Majidinia, Mojtaba Kaviani, Mehdi Yousefi, Bahman Yousefi.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract The cancer stem cell (CSC) model encompasses an advantageous paradigm that in recent decades provides a better elucidation for many important biological aspects of cancer initiation, progression, metastasis, and, more important, development of multidrug resistance (MDR). Such several other hematological malignancies and solid tumors and the identification and isolation of ovarian cancer stem cells (OV‐CSCs) show that ovarian cancer also follows this hierarchical model. Gaining a better insight into CSC‐mediated resistance holds promise for improving current ovarian cancer therapies and prolonging the survival of recurrent ovarian cancer patients in the future. Therefore, in this review, we will discuss some important mechanisms by which CSCs can escape chemotherapy, and then review the recent and growing body of evidence that supports the contribution of CSCs to MDR in ovarian cancer. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.26768   open full text
• Induction of apoptotic but not autophagic cell death by Cinnamomum cassia extracts on human oral cancer cells.
  Ching‐Han Yu, Shu‐Chen Chu, Shun‐Fa Yang, Yih‐Shou Hsieh, Chih‐Yi Lee, Pei‐Ni Chen.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nCinnamomum cassia has been widely studied in different fields to reveal its antidiabetic, antidepressive, antiviral, anti‐inflammatory, antiosteoporotic, and anticancer effects. Its antimalignant activities have been explored in lung cancer, breast cancer, colorectal cancer, and even oral cancer, but the detailed signaling mechanism and effects of this plant on animal models need to be clarified. In the current study, \nC. cassia extract (CCE) was used to investigate the antitumorigenesis mechanism in vitro and in vivo. The major constituents of CCE used in this study were coumarin, cinnamic acid, and cinnamic aldehyde. CCE reduced the viability, number, and colony formation of human oral cancer cells, and induced their apoptosis. Caspase‐3 activation, Bcl‐2 reduction, and phosphatidylserine inversion were involved in CCE‐stimulated apoptosis. CCE also enhanced the expression of autophagic markers, including acidic vesicular organelle, microtubule‐associated protein 1 light chain 3‐I, autophagy‐related protein 14, rubicon, and p62. The combined treatment of CCE and caspase inhibitor significantly restored mitochondrial membrane potential (Δ\nψ\nm) and cell viability. However, the combined treatment of CCE and autophagy inhibitor further reduced the cell viability indicating that autophagy might be a survival pathway of CCE‐treated SASVO3 cells. In contrast, CCE treatment for 12 days did not adversely affect SASVO3 tumor‐bearing nude mice. CCE also elicited dose‐dependent effects on the decrease in tumor volume, tumor weight, and Ki‐67 expression. These results suggested that CCE showed the potential for the complementary treatment of oral caner." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27338   open full text
• Resveratrol restores sensitivity of glioma cells to temozolamide through inhibiting the activation of Wnt signaling pathway.
  Hua‐Chao Yang, Jun‐Yi Wang, Xing‐Yao Bu, Bin Yang, Bang‐Qing Wang, Sen Hu, Zhao‐Yue Yan, Yu‐Shuai Gao, Shuang‐Yin Han, Ming‐Qi Qu.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nMalignant gliomas are aggressive primary neoplasms that originate in the glial cells of the brain or the spine with notable resistance to standard treatment options. We carried out the study with the aim to shed light on the sensitization of resveratrol to temozolomide (TMZ) against glioma through the Wnt signaling pathway. Initially, glioma cell lines with strong resistance to TMZ were selected by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay. Then, the glioma cells were subjected to resveratrol, TMZ, Wnt signaling pathway inhibitors, and activators. Cell survival rate and inhibitory concentration at half maximum value were detected by MTT, apoptosis by flow cytometry, and terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling staining, in vitro proliferation by hanging drop method and β‐catenin translocation into nuclei by TOP/FOP‐FLASH assay. The expressions of the Wnt signaling pathway‐related and apoptosis‐related factors were determined by western blot analysis. Nude mice with glioma xenograft were established to detect tumorigenic ability. Glioma cell lines T98G and U138 which were highly resistant to TMZ were selected for subsequent experiments. Resveratrol increased the efficacy of TMZ by restraining cell proliferation, tumor growth, and promoting cell apoptosis in glioma cells. Resveratrol inhibited Wnt2 and β‐catenin expressions yet elevated GSK‐3β expression. Moreover, the Wnt signaling pathway participates in the sensitivity enhancing of resveratrol to TMZ via regulating \nO\n6‐methylguanine‐DNA methyltransferase (MGMT) expression. Resveratrol sensitized TMZ‐induced glioma cell apoptosis by repressing the activation of the Wnt signaling pathway and downregulating MGMT expression, which may confer new thoughts to the chemotherapy of glioma." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27409   open full text
• Probiotics importance and their immunomodulatory properties.
  Bahman Yousefi, Majid Eslami, Abdolmajid Ghasemian, Parviz Kokhaei, Amir Salek Farrokhi, Narges Darabi.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Mammalian intestine contains a large diversity of commensal microbiota, which is far more than the number of host cells. Probiotics play an insecure and protective role against the colonization of intestinal pathogenic microbes and increase mucosal integrity by stimulating epithelial cells. Probiotics have innate capabilities in many ways, including receptor antagonism, receptor expression, binding and expression of adapter proteins, expression of negative regulatory signal molecules, induction of microRNAs, endotoxin tolerance, and ultimately secretion of immunomodulatory proteins, lipids, and metabolites to modulate the immune system. Probiotic bacteria can affect homeostasis, inflammation, and immunopathology through direct or indirect effects on signaling pathways as immunosuppressant or activators. Probiotics suppress inflammation by inhibiting various signaling pathways such as the nuclear factor‐κB (NF‐κβ) pathway, possibly related to alterations in mitogen‐activated protein kinases and pattern recognition receptors pathways. Probiotics can also inhibit the binding of lipopolysaccharides to the CD14 receptor, thereby reducing the overall activation of NF‐κβ and producing proinflammatory cytokines. Some effects of modulation by probiotics include cytokine production by epithelial cells, increased mucin secretion, increased activity of phagocytosis, and activation of T and natural killer T cells, stimulation of immunoglobulin A production and decreased T cell proliferation. Intestinal microbiota has a major impact on the systemic immune system. Specific microbiota controls the differentiation of cells in lamina propria, in which Th17 cells secrete interleukin 17. The presence of Th17 and Treg cells in the small intestine is associated with intestinal microbiota, with the preferential Treg differentiation and the absence of Th17 cells, possibly reflecting alterations in the lamina propria cytokines and the intestinal gut microbiota. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27559   open full text
• SEMA3B‐AS1‐inhibited osteogenic differentiation of human mesenchymal stem cells revealed by quantitative proteomics analysis.
  Chen Zhang, Yun Zhu, Yugang Liu, Xiguang Zhang, Qiaoning Yue, Li Li, Yatang Chen, Sheng Lu, Zhaowei Teng.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Human mesenchymal stem cells (hMSCs) are fibroblastoid multipotent adult stem cells with capacities of differentiation into osteoblasts and chondrocytes and show great potential in new bone formation and bone repair‐related clinical settings, such as osteoporosis. Long noncoding RNAs (lncRNAs) have been demonstrated to play important roles in various biological processes. Here, we report an antisense lncRNA SEMA3B‐AS1 regulating hMSCs osteogenesis. SEMA3B‐AS1 is proximal to a member of the semaphorin family Sema3b. Overexpression of SEMA3B‐AS1 using the lentivirus system markedly inhibits the proliferation of hMSCs and meanwhile reduces osteogenic differentiation. Using a comprehensive proteomic technique named isobaric tag for relative and absolute quantitation, we found that SEMA3B‐AS1 significantly alters the process of osteogenesis through downregulating the expression of proteins involved in actin cytoskeleton, focal adhesion, and extracellular matrix–receptor interaction, while increasing the expression of proteins in the spliceosome. Collectively, we find that SEMA3B‐AS1 is a target for controlling osteogenesis of hMSCs. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.26776   open full text
• EA15, MIR22, LINC00472 as diagnostic markers for diabetic kidney disease.
  Yan‐Zhe Wang, Ding‐Yu Zhu, Xin‐Miao Xie, Miao Ding, Yong‐Lan Wang, Lin‐Lin Sun, Nan Zhang, E. Shen, Xiao‐Xia Wang.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nThis study aimed to investigate the molecular mechanisms of diabetic kidney disease (DKD) and to explore new potential therapeutic strategies and biomarkers for DKD. First we analyzed the differentially expressed changes between patients with DKD and the control group using the chip data in Gene Expression Omnibus (GEO) database. Then the gene chip was subjected to be annotated again, so as to screen long noncoding RNAs (lncRNAs) and study expression differences of these lncRNAs in DKD and controlled samples. At last, the function of the differential lncRNAs was analyzed. A total of 252 lncRNAs were identified, and 14 were differentially expressed. In addition, there were 1,629 differentially expressed messenger RNAs (mRNAs) genes, and proliferation and apoptosis adapter protein 15 (PEA15), \nMIR22, and long intergenic nonprotein coding RNA 472 (\nLINC00472) were significantly differentially expressed in DKD samples. Through functional analysis of the encoding genes coexpressed by the three lncRNAs, we found these genes were mainly enriched in type 1 diabetes and autoimmune thyroid disease pathways, whereas in Gene Ontology (GO) function classification, they were also mainly enriched in the immune response, type I interferon signaling pathways, interferon‐γ mediated signaling pathways, and so forth. To summary, we identified \nEA15, \nMIR22, and \nLINC00472 may serve as the potential diagnostic markers of DKD." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27539   open full text
• Integrated bioinformatic analysis reveals YWHAB as a novel diagnostic biomarker for idiopathic pulmonary arterial hypertension.
  Tao Wang, Xuan Zheng, Ruidong Li, Xintian Liu, Jinhua Wu, Xiaodan Zhong, Wenjun Zhang, Yujian Liu, Xingwei He, Wanjun Liu, Hongjie Wang, Hesong Zeng.
  Journal of Cellular Physiology. October 14, 2018
  --- - "\nAbstract\nIdiopathic pulmonary arterial hypertension (IPAH) is a severe cardiovascular disease that is a serious threat to human life. However, the specific diagnostic biomarkers have not been fully clarified and candidate regulatory targets for IPAH have not been identified. The aim of this study was to explore the potential diagnostic biomarkers and possible regulatory targets of IPAH. We performed a weighted gene coexpression network analysis and calculated module‐trait correlations based on a public microarray data set (GSE703) and six modules were found to be related to IPAH. Two modules which have the strongest correlation with IPAH were further analyzed and the top 10 hub genes in the two modules were identified. Furthermore, we validated the data by quantitative real‐time polymerase chain reaction (qRT‐PCR) in an independent sample set originated from our study center. Overall, the qRT‐PCR results were consistent with most of the results of the microarray analysis. Intriguingly, the highest change was found for YWHAB, a gene encodes a protein belonging to the 14‐3‐3 family of proteins, members of which mediate signal transduction by binding to phosphoserine‐containing proteins. Thus, \nYWHAB was subsequently selected for validation. In congruent with the gene expression analysis, plasma 14‐3‐3β concentrations were significantly increased in patients with IPAH compared with healthy controls, and 14‐3‐3β expression was also positively correlated with mean pulmonary artery pressure (\nR\n2 = 0.8783; \np < 0.001). Taken together, using weighted gene coexpression analysis, \nYWHAB was identified and validated in association with IPAH progression, which might serve as a biomarker and/or therapeutic target for IPAH." - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27381   open full text
• Diagnostic, prognostic, and therapeutic potency of microRNA 21 in the pathogenesis of colon cancer, current status and prospective.
  Amirhossein Bahreyni, Melika Rezaei, Afsane Bahrami, Majid Khazaei, Hamid Fiuji, Mikhail Ryzhikov, Gordon A. Ferns, Amir Avan, Seyed Mahdi Hassanian.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Aberrant microRNA (miR) expression is implicated in multiple human malignancies. miR‐21, acting as a proto‐oncogene, is involved in a variety of cellular processes and tumorigenesis and is frequently overexpressed in some cancer types. Several tumor suppressors, metastatic, and apoptotic genes have been identified as miR‐21 targets, including Ras homolog gene family member B, PTEN, Sprouty2, programmed cell death 4, Integrin‐β4, and E‐cadherin thereby regulating tumor growth, invasion, and metastasis. There is a growing evidence that miR‐21 expression is associated with clinical outcomes in patients with colorectal cancer (CRC). In this review, we summarize the potential diagnostic, prognostic, and therapeutic values of miR‐21 in CRC progression for a better understanding and hence a better management of this disease. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27580   open full text
• Snake venoms promote stress‐induced senescence in human fibroblasts.
  Anna Lewinska, Aleksandra Bocian, Vladimir Petrilla, Jagoda Adamczyk‐Grochala, Karolina Szymura, Wiktoria Hendzel, Edyta Kaleniuk, Konrad K. Hus, Monika Petrillova, Maciej Wnuk.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Snake venoms are widely studied in terms of their systemic toxicity and proteolytic, hemotoxic, neurotoxic, and cytotoxic activities. However, little is known about snake‐venom‐mediated effects when used at low, noncytotoxic concentrations. In the current study, two human fibroblast cell lines of different origin, namely WI‐38 fetal lung fibroblasts and BJ foreskin fibroblasts were used to investigate snake‐venom‐induced adaptive response at a relatively noncytotoxic concentration (0.01 µg/ml). The venoms of Indochinese spitting cobra ( Naja siamensis), western green mamba ( Dendroaspis viridis), forest cobra ( Naja melanoleuca), and southern copperhead ( Agkistrodon contortrix) were considered. Snake venoms promoted FOXO3a‐mediated oxidative stress response and to a lesser extent DNA damage response, which lead to changes in cell cycle regulators both at messenger RNA and protein levels, limited cell proliferation and migration, and induced cellular senescence. Taken together, we have shown for the first time that selected snake venoms may also exert adverse effects when used at relatively noncytotoxic concentrations. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27382   open full text
• The vacuolated morphology of chordoma cells is dependent on cytokeratin intermediate filaments.
  Lauren Resutek, Adam H. Hsieh.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Notochordal cells (NCs), characterized by their vacuolated morphology and coexpression of cytokeratin and vimentin intermediate filaments (IFs), form the immature nucleus pulposus (NP) of the intervertebral disc. As humans age, NCs give way to mature NP cells, which do not possess a vacuolated morphology and typically only express vimentin IFs. In light of their concomitant loss, we investigated the relationship between cytosolic vacuoles and cytokeratin IFs, specifically those containing cytokeratin‐8 proteins, using a human chordoma cell line as a model for NCs. We demonstrate that the chemical disruption of IFs with acrylamide, F‐actin with cytochalasin‐D, and microtubules with nocodazole all result in a significant (p < 0.001) decrease in vacuolation. However, vacuole loss was the greatest in acrylamide‐treated cells. Examination of the individual roles of vimentin and cytokeratin‐8 IFs in the existence of vacuoles was accomplished using small interfering RNA–mediated RNA interference to knock down either vimentin or cytokeratin‐8 expression. Reduction of cytokeratin‐8 expression was associated with a less‐vacuolated cell morphology. These data demonstrate that cytokeratin‐8 IFs are involved in stabilizing vacuoles and that their diminished expression could play a role in the loss of vacuolation in NCs during aging. A better understanding of the NCs may assist in preservation of this cell type for NP maintenance and regeneration. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.26809   open full text
• LINC01433 promotes hepatocellular carcinoma progression via modulating the miR‐1301/STAT3 axis.
  Haijin Huang, Yan‐Zhi Bu, Xiao‐Yu Zhang, Juan Liu, Li‐Yao Zhu, Yong Fang.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Long noncoding RNAs (lncRNAs) have been demonstrated to play significant roles in hepatocellular carcinoma (HCC) tumor progression. LINC01433 has been implicated in the progression of lung cancer. However, its biological role in HCC remains poorly understood. In our current study, we focused on the detailed mechanism of LINC01433 in HCC development. First, it was exhibited that LINC01433 was remarkably elevated in HCC cells, which indicated that LINC01433 was involved in HCC. Then, knockdown of LINC01433 was able to restrain HCC cell proliferation and cell colony formation and greatly induced cell apoptosis. On the contrary, overexpression of LINC01433 promoted HCC cell proliferation, increased cell colony formation, and enhanced cell invasion capacity. Subsequently, we found that miR‐1301 was remarkably decreased in HCC cells, and it can serve as a target of LINC01433 according to bioinformatics analysis. In addition, the binding correlation between them was validated by performing RNA pull‐down experiments and RIP assay. Moreover, STAT3 was predicted and validated as a target of miR‐1301, and it was shown that miR‐1301 mimics significantly suppressed STAT3 in HCC cells. Finally, in vivo models were established, and the results demonstrated that silencing of LINC01433 could repress HCC development through modulating miR‐1301 and STAT3. Taken together, these results indicated in our study that LINC01433 participated in HCC progression through modulating the miR‐1301/STAT3 axis and it might act as a novel biomarker in HCC diagnosis and treatment. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27366   open full text
• Interferon‐stimulated gene 15 enters posttranslational modifications of p53.
  Yang Wang, Qi Ding, Yu‐Chen Lu, Shi‐Yang Cao, Qing‐Xue Liu, Lei Zhang.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2 Abstract The tumor suppressor protein p53 is a central governor of various cellular signals. It is well accepted that ubiquitination as well as ubiquitin‐like (UBL) modifications of p53 protein is critical in the control of its activity. Interferon‐stimulated gene 15 (ISG15) is a well‐known UBL protein with pleiotropic functions, serving both as a free intracellular molecule and as a modifier by conjugating to target proteins. Initially, attentions have historically focused on the antiviral effects of ISG15 pathway. Remarkably, a significant role in the processes of autophagy, DNA repair, and protein translation provided considerable insight into the new functions of ISG15 pathway. Despite the deterministic revelation of the relation between ISG15 and p53, the functional consequence of p53 ISGylation appears somewhat confused. More important, more recent studies have hinted p53 ubiquitination or other UBL modifications that might interconnect with its ISGylation. Here, we aim to summarize the current knowledge of p53 ISGylation and the differences in other significant modifications, which would be beneficial for the development of p53‐based cancer therapy. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27347   open full text
• Tyrosine phosphorylation of HDAC3 by Src kinase mediates proliferation of HER2‐positive breast cancer cells.
  Jaesung Seo, Garam Guk, Seung‐Ho Park, Mi‐Hyeon Jeong, Ji‐Hoon Jeong, Ho‐Geun Yoon, Kyung‐Chul Choi.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract The role of histone deacetylase 3 (HDAC3) is to repress the expression of various genes by eliminating acetyl group from histone. Thus, the regulation of HDAC3 activity is essential to maintain cellular homeostasis. In this study, we found that HDAC3 interacts with c‐Src kinase. However, the interaction between HDAC3 and c‐Src was previously reported, it has still been ambiguous whether c‐Src phosphorylates HDAC3 and affects the function of HDAC3. First, we confirmed that HDAC3 directly binds to c‐Src, and c‐Src identified to interact with C‐terminal domain (277–428 a.a.) of HDAC3. c‐Src also phosphorylated three tyrosine sites of HDAC3 at tyrosine 325, 328, and 331. Importantly, wild‐type c‐Src increases HDAC3 activity, but not mutant c‐SrcK298M (kinase inactive form). When these tyrosine residues are all substituted for alanine residues, the deacetylase activity of mutant HDAC3 was abolished. In addition, a proliferation of HER2‐positive breast cancer cells expressing phosphorylation deficient mutant HDAC3 is decreased in comparison with control cells. Thus, our findings suggested that phosphorylation of HDAC3 by c‐Src kinase regulates the HDAC3 activity and the proliferation of breast cancer cells. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27378   open full text
• Cinnamaldehyde regulates H2O 2‐induced skeletal muscle atrophy by ameliorating the proteolytic and antioxidant defense systems.
  Nirmaljeet Kaur, Prachi Gupta, Vikram Saini, Sandeep Sherawat, Sanjeev Gupta, Anita Dua, Vinod Kumar, Elisha Injeti, Ashwani Mittal.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Skeletal muscle atrophy/wasting is associated with impaired protein metabolism in diverse physiological and pathophysiological conditions. Elevated levels of reactive oxygen species (ROS), disturbed redox status, and weakened antioxidant defense system are the major contributing factors toward atrophy. Regulation of protein metabolism by controlling ROS levels and its associated catabolic pathways may help in treating atrophy and related clinical conditions. Although cinnamaldehyde (CNA) enjoys the established status of antioxidant and its role in ROS management is reported, impact of CNA on skeletal muscle atrophy and related pathways is still unexplored. In the current study, the impact of CNA on C2C12 myotubes and the possible protection of cultured cells from H 2O 2‐induced atrophy is examined. Myotubes were treated with H 2O 2 in the presence and absence of CNA and the changes in the antioxidative, proteolytic systems, and mitochondrial functions were scored. Morphological analysis showed significant protective effects of CNA on length, diameter, and nuclei fusion index of myotubes. The evaluation of biochemical markers of atrophy; creatine kinase, lactate dehydrogenase, succinate dehydrogenase along with the study of muscle‐specific structural protein (i.e., myosin heavy chain‐fast [MHCf] type) showed significant protection of proteins by CNA. CNA pretreatment not only checked the activation of proteolytic systems (ubiquitin‐proteasome E3‐ligases [MuRF1/Atrogin1]), autophagy [Beclin1/LC3B], cathepsin L, calpain, caspase), but also prevented any alteration in the activities of antioxidative defense enzymes (catalase, glutathione‐ S‐transferase, glutathione‐peroxidase, superoxide dismutase, glutathione reductase). The results suggest that CNA protects myotubes from H 2O 2‐induced atrophy by inhibiting/resisting the amendments in proteolytic systems and maintains cellular redox‐balance. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27348   open full text
• Exploring and comparing of the gene expression and methylation differences between lung adenocarcinoma and squamous cell carcinoma.
  Yang Yang, Meng Wang, Bao Liu.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Lung cancer is one of the most frequently diagnosed malignant tumors and the main reason for cancer‐related death around the world, whereas nonsmall cell lung cancer that consists two subtypes: lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) is responsible for an estimated 85% of all lung cancers. The current study aimed to explore gene expression and methylation differences between LUAD and LUSC. EdgeR was used to identify differentially regulated genes between normal and cancer in the LUAD and LUSC extracted from The Cancer Genome Atlas (TCGA), respectively, whereas SAM was used to find genes with differential methylation between normal and cancer in the LUAD and LUSC, respectively. Finally, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to analyze the function which these genes enriched in. A total of 391 genes with opposite methylation patterns in LUAD and LUSC and four functional pathways were obtained (false discovery rate (FDR) < 0.1). These pathways mainly included fat digestion and absorption, phenylalanine metabolism, bile secretion, and so on, which were related to the airframe nutrition metabolic pathway. Moreover, two genes CTSE (cathepsin E) and solute carrier family 5 member 7 (SLC5A7) were also found, among which CTSE was overexpressed and hypomethylated in LUAD corresponding to normal lung tissues, whereas SLC5A7 showed the opposite in LUAD. In conclusion, this study investigated the differences between the gene expression and methylation patterns in LUAD and LUSC, and explored their different biological characteristics. Further understanding of these differences may promote the discovery and development of new, accurate strategies for the prevention, diagnosis, and treatment of lung cancer. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27240   open full text
• MiR‐208b regulates cell cycle and promotes skeletal muscle cell proliferation by targeting CDKN1A.
  Jian Wang, Chengchuang Song, Xiukai Cao, Hui Li, Hanfang Cai, Yilei Ma, Yongzhen Huang, Xianyong Lan, Chuzhao Lei, Yun Ma, Yueyu Bai, Fengpeng Lin, Hong Chen.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Skeletal muscle is the most abundant tissue in the body. The development of skeletal muscle cell is complex and affected by many factors. A sea of microRNAs (miRNAs) have been identified as critical regulators of myogenesis. MiR‐208b, a muscle‐specific miRNA, was reported to have a connection with fiber type determination. However, whether miR‐208b has effect on proliferation of muscle cell was under ascertained. In our study, cyclin‐dependent kinase inhibitor 1A (CDKN1A), which participates in cell cycle regulation, was predicted and then validated as one target gene of miR‐208b. We found that overexpression of miR‐208b increased the expression of cyclin D1, cyclin E1, and cyclin‐dependent kinase 2 at the levels of messenger RNA and protein in cattle primary myoblasts in vivo and in vitro. Flow cytometry showed that forced expression of miR‐208b increased the percentage of cells at the S phase and decreased the percentage of cells at the G0/G1 phase. These results indicated that miR‐208b participates in the cell cycle regulation of cattle primary myoblast cells. 5‐Ethynyl‐20‐deoxyuridine and 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assays showed that overexpression of miR‐208b promoted the proliferation of cattle primary myoblasts. Therefore, we conclude that miR‐208b participates in the cell cycle and proliferation regulation of cattle primary skeletal muscle cell through the posttranscriptional downregulation of CDKN1A. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27146   open full text
• Alteration of cellular and immune‐related properties of bone marrow mesenchymal stem cells and macrophages by K562 chronic myeloid leukemia cell derived exosomes.
  Nazli Jafarzadeh, Zohreh Safari, Majid Pornour, Naser Amirizadeh, Mehdi Forouzandeh Moghadam, Majid Sadeghizadeh.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract Leukemic cells can impact the bone marrow niche to create a tumor‐favorable microenvironment using their secreted factors. Little knowledge is available about immunosuppressive and tumor‐promoting properties of chronic myeloid leukemia derived exosomes in bone marrow stromal components. We report here that K562‐derived exosomes can affect the gene expression, cytokine secretion, nitric oxide (NO) production, and redox potential of bone marrow mesenchymal stem cells (BM‐MSCs) and macrophages. Human BM‐MSCs and mouse macrophages were treated with K562‐derived exosomes. Our results demonstrated that the expression of the genes involved in hematopoietic developmental pathways and immune responses, including C‐X‐C motif chemokine 12 (Cxcl12), Dickkopf‐related protein 1 (DKK1), wnt5a, interleukin 6 (IL‐6), transforming growth factor‐beta, and tumor necrosis factor‐alpha (TNF‐alpha), changed with respect to time and exosome concentration in BM‐MSCs. The TNF‐alpha level was higher in exosome‐treated BM‐MSCs compared with the control. Exosome treatment of BM‐MSCs led to an increased production of NO and a decreased production of reactive oxygen species (ROS) in a time‐ and concentration‐dependent manner. We have shown that K562‐derived exosomes induce overexpression of IL‐10 and TNF‐alpha and downregulation of iNOS transcript levels in macrophages. The enzyme‐linked immunosorbent assay results showed that TNF‐alpha and IL‐10 secretions increased in macrophages. Treatment of macrophages with purified exosomes led to reduced NO and ROS levels. These results suggest that K562‐derived exosomes may alter the local bone marrow niche toward a leukemia‐reinforcing microenvironment. They can modulate the inflammatory molecules (TNF‐alpha and NO) and the redox potential of BM‐MSCs and macrophages and direct the polarization of macrophages toward tumor‐associated macrophages. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27142   open full text
• MicroRNA‐133b suppresses bladder cancer malignancy by targeting TAGLN2‐mediated cell cycle.
  Feng Zhao, Liu‐Hua Zhou, Yu‐Zheng Ge, Wen‐Wen Ping, Xin Wu, Zhong‐Le Xu, Min Wang, Zuo‐Liang Sha, Rui‐Peng Jia.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- Abstract MicroRNAs (miRNAs), a group of small noncoding RNAs, are widely involved in the regulation of gene expression via binding to complementary sequences at 3′‐untranslated regions (3′‐UTRs) of target messenger RNAs. Recently, downregulation of miR‐133b has been detected in various human malignancies. Here, the potential biological role of miR‐133b in bladder cancer (BC) was investigated. In this study, we found the expression of miR‐133b was markedly downregulated in BC tissues and cell lines (5637 and T24), and was correlated with poor overall survival. Notably, transgelin 2 (TAGLN2) was found to be widely upregulated in BC, and overexpression of TAGLN2 also significantly increased risks of advanced TMN stage. We further identified that upregulation of miR‐133b inhibited glucose uptake, invasion, angiogenesis, colony formation and enhances gemcitabine chemosensitivity in BC cell lines by targeting TAGLN2. Additionally, we showed that miR‐133b promoted the proliferation of BC cells, at least partially through a TAGLN2‐mediated cell cycle pathway. Our results suggest a novel miR‐133b/TAGLN2/cell cycle pathway axis controlling BC progression; a molecular mechanism which may offer a potential therapeutic target. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27288   open full text
• Arsenic trioxide inhibits the differentiation of fibroblasts to myofibroblasts through nuclear factor erythroid 2‐like 2 (NFE2L2) protein and the Smad2/3 pathway.
  Lingzhi Zhong, Haojie Hao, Deyun Chen, Qian Hou, Ziying Zhu, Wenjun He, Sujing Sun, Mengli Sun, Meirong Li, Xiaobing Fu.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2 Background Tissue contraction and the extracellular matrix deposition are part of the pathogenesis of hypertrophic scars. The transcriptional factor NFE2L2 inhibits fibroblast differentiation in idiopathic pulmonary fibrosis and promotes myofibroblast dedifferentiation. Our previous study showed that the transcription factor NFE2L2 was strongly induced on treatment with arsenic trioxide (ATO). Objective The present study sought to investigate the effect of ATO on myofibroblast formation to determine its potential role in hypertrophic scar treatment. Methods Small interfering RNA against NFE2L2 was used on treatment with ATO in human skin myofibroblasts. The expression levels of fibrosis markers were assessed by reverse transcription polymerase chain reaction, western blot, and immunofluorescence staining. The transforming growth factor‐β1 (TGF‐β1)/Smad2/3 signaling was detected by western blot. A rabbit ear model was used to evaluate the antifibrotic role of ATO. Results At the cellular level, ATO abolished fibroblast differentiation in response to TGF‐β1. ATO reduced TGF‐β1‐induced reactive oxygen species accumulation through increased expression of the antioxidant gene HO‐1 in fibroblasts. In addition, ATO promoted the nuclear translocation of NFE2L2 and inhibited the phosphorylation of Smad2/3. In the rabbit ear model, ATO prevented the progression of hypertrophic scar formation. Conclusions This study provides the first evidence implying that ATO inhibits the formation of myofibroblasts in vivo and in vitro and provides a possible treatment for hypertrophic scars. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27073   open full text
• Peroxisome proliferator‐activated receptor γ modulates renal crystal retention associated with high oxalate concentration by regulating tubular epithelial cellular transdifferentiation.
  Shujue Li, Yu Lan, Wenzheng Wu, Xiaolu Duan, Zhenzhen Kong, Wenqi Wu, Guohua Zeng.
  Journal of Cellular Physiology. October 14, 2018
  --- - |2- The differentiated phenotype of renal tubular epithelial cell exerts significant effect on crystal adherence. Peroxisome proliferator‐activated receptor γ (PPARγ) has been shown to be critical for the regulation of cell transdifferentiation in many physiological and pathological conditions; however, little is known about its role in kidney stone formation. In the current study, we found that temporarily high oxalate concentration significantly decreased PPARγ expression, induced Madin Darby Canine Kidney cell dedifferentiation, and prompted subsequent calcium oxalate (CaOx) crystal adhesion in vitro. Furthermore, cell redifferentiation after the removal of the high oxalate concentration, along with a decreasing affinity to crystals, was an endogenic PPARγ‐dependent process. In addition, the PPARγ antagonist GW9662, which can depress total‐PPARγ expression and activity, enhanced cell dedifferentiation induced by high oxalate concentration and inhibited cell redifferentiation after removal of the high oxalate concentration. These effects were partially reversed by the PPARγ agonist 15d‐PGJ2. Similar results were observed in animals that suffered from temporary hyperoxaluria followed by a recovery period. The active crystal‐clearing process occurs through the transphenotypical morphology of renal tubular epithelial cells, reflecting cell transdifferentiation during the recovery period. However, GW9662 delayed cell redifferentiation and increased the secondary temporary crystalluria‐induced crystal retention. This detrimental effect was partially reversed by 15d‐PGJ2. Taken together, our results revealed that endogenic PPARγ activity plays a vital regulatory role in crystal clearance, subsequent crystal adherence, and CaOx stone formation via manipulating the transdifferentiation of renal tubular epithelial cells. - Journal of Cellular Physiology, EarlyView.
  

  October 14, 2018   doi: 10.1002/jcp.27102   open full text
• Cover Image, Volume 233, Number 12, December 2018.
  May Chammaa, Agnes Malysa, Carlos Redondo, Hyejeong Jang, Wei Chen, Gerold Bepler, Rodrigo Fernandez‐Valdivia.
  Journal of Cellular Physiology. October 12, 2018
  --- - - Journal of Cellular Physiology, Volume 233, Issue 12, Page i-i, December 2018.
  

  October 12, 2018   doi: 10.1002/jcp.27479   open full text
• Table of Contents, Editor's Choice, Highlights.
  
  Journal of Cellular Physiology. October 12, 2018
  --- - - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9031-9042, December 2018.
  

  October 12, 2018   doi: 10.1002/jcp.27478   open full text
• DL‐3‐n‐butylphthalide alleviates vascular cognitive impairment by regulating endoplasmic reticulum stress and the Shh/Ptch1 signaling‐pathway in rats.
  Xiao‐Li Niu, Xin Jiang, Guo‐Dong Xu, Gui‐Min Zheng, Zhi‐Peng Tang, Nan Yin, Xiu‐Qin Li, Yan‐Yan Yang, Pei‐Yuan Lv.
  Journal of Cellular Physiology. October 11, 2018
  --- - |2 Abstract Background DL‐3‐n‐butylphthalide (NBP) has been approved to be effective in improving cognitive deficits. The aim of the current study was to determine whether NBP protects against cognitive deficits in a rat model of vascular dementia (VD) induced by chronic cerebral hypoperfusion (CCH) by regulating the sonic hedgehog (Shh)/patched1 (Ptch1) pathway and endoplasmic reticulum stress (ERS)‐related markers. Methods Adult male Sprague‐Dawley rats were subjected to permanent bilateral occlusion of the common carotid arteries (2VO) to established the model of VD. These rats were randomly divided into five groups: sham, model, NBP30 (30 mg/kg), NBP 60 (60 mg/kg), and NBP 120 (120 mg/kg) groups. The Morris water maze test was used to assess for cognitive function at 4 weeks after operation. Results NBP significantly alleviated spatial learning and memory impairment, and inhibited the loss of neurons in the CA1 region of the hippocampus. Western blot analysis and real‐time quantitative polymerase chain reaction analysis revealed that plasticity‐related synaptic markers and the Shh/Ptch1 pathway significantly increased in the NBP treated groups, while ERS‐related markers decreased. Conclusion The results of the current study prove that the Shh/Ptch1 pathway plays an essential role in the model of VD. NBP had protective effects on cognitive impairment induced by CCH. This mechanism was associated with ERS and the Shh/Ptch1 pathway. Meanwhile, the Shh/Ptch1 pathway and ERS may interact with each other. - Journal of Cellular Physiology, EarlyView.
  

  October 11, 2018   doi: 10.1002/jcp.27332   open full text
• Chrysin leads to cell death in endometriosis by regulation of endoplasmic reticulum stress and cytosolic calcium level.
  Soomin Ryu, Fuller W. Bazer, Whasun Lim, Gwonhwa Song.
  Journal of Cellular Physiology. October 10, 2018
  --- - |2- Abstract Chrysin is a natural compound derived from honey, propolis, or passion flowers and has many functional roles, such as antiinflammatory and antiangiogenesis effects. Although endometriosis is a benign gynecological disease, there is a need to identify the pathology and develop a therapy for endometriosis. Elucidating the biological mechanism of chrysin on endometriosis will improve the understanding of endometriosis. In this study, we confirmed the apoptotic effects of chrysin in human endometriotic cells using End1/E6E7 (endocervix‐derived endometriotic cells) and VK2/E6E7 (vaginal mucosa‐derived epithelial endometriotic cells). The results showed that chrysin suppressed the proliferation of endometriosis and induced programmed cell death through changing the cell cycle proportion and increasing the cytosolic calcium level and generation of reactive oxygen species. In addition, chrysin activated endoplasmic reticulum (ER) stress by stimulating the unfolded protein response proteins, especially the 78‐kDa glucose‐regulated protein–PRKR‐like ER kinase (PERK)–eukaryotic translation initiation factor 2α (eIF2α) pathway in both endometriotic cell lines. Furthermore, chrysin inactivated the intracellular phosphoinositide 3‐kinase (PI3K)/protein kinase B (PKB, also known as AKT) signaling pathway in a dose‐dependent manner. Collectively, the results of this study indicated that chrysin induced programmed cell death by activating the ER stress response and inactivating the PI3K signaling pathways in human endometriotic cells. - Journal of Cellular Physiology, EarlyView.
  

  October 10, 2018   doi: 10.1002/jcp.26770   open full text
• Long noncoding RNA Gm6135 functions as a competitive endogenous RNA to regulate toll‐like receptor 4 expression by sponging miR‐203‐3p in diabetic nephropathy.
  Ting‐Ting Ji, Ya‐Kun Wang, Ying‐Chun Zhu, Cong‐Pu Gao, Xiao‐Ying Li, Ji Li, Feng Bai, Shou‐Jun Bai.
  Journal of Cellular Physiology. October 08, 2018
  --- - |2- Abstract We aim to explore the relationship between Gm6135 and diabetic nephropathy. We detected the relative expression levels of Gm6135 and toll‐like receptor 4 (TLR4) in diabetic nephropathy mice and high‐glucose‐cultured mouse mesangial cells SV40‐MES‐13 by the quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) and western blot detection. Cell proliferation and apoptosis were detected after small interfering RNA (siRNA) interference or plasmid overexpression of Gm6135/TLR4, and bioinformatics method was used to predict and screen miR‐203 as an intermediate factor. Through dual‐luciferase reporter gene, RNA pull‐down, qRT‐PCR, and western blot, the binding relationship between Gm6135, miR‐203‐3p, and TLR4 was confirmed. The possibility of the competing endogenous RNA mechanism was demonstrated by cell localization assays and rip assays. Finally, the proliferation of mouse mesangial cells SV40‐MES‐13 was detected after mimics and inhibitor of microRNA, which were reversed with TLR4 overexpression and siRNA. The results showed that the relative expression levels of Gm6135 and TLR4 in the kidney and high‐glucose‐cultured mouse mesangial cells of diabetic nephropathy mice increased significantly. Overexpression or downregulation of Gm6135/TLR4 significantly affected the proliferation and apoptosis of mouse mesangial cells. Gm6135 upregulates TLR4 by competitively binding to miR‐203‐3p. - Journal of Cellular Physiology, EarlyView.
  

  October 08, 2018   doi: 10.1002/jcp.27412   open full text
• The imbalance of Th17/Treg axis involved in the pathogenesis of preeclampsia.
  Shadi Eghbal‐Fard, Mehdi Yousefi, Hanieh Heydarlou, Majid Ahmadi, Simin Taghavi, Aliakbar Movasaghpour, Farhad Jadidi‐Niaragh, Bahman Yousefi, Sanam Dolati, Mohammad Hojjat‐Farsangi, Reza Rikhtegar, Mohammad Nouri, Leili Aghebati‐Maleki.
  Journal of Cellular Physiology. October 02, 2018
  --- - "\nAbstract\n\nProblem\nInappropriate activation of the immune system, particularly the imbalance of T‐helper type 17 (Th17)/regulatory T (Treg) cells is thought to play considerable roles in preeclampsia (PE). To investigate the probable effects of the adaptive immune system in the pathophysiology of PE, we analyzed the dynamic changes of Th17/Treg cells, cytokines profile, and transcription pattern of Th17/Treg‐related genes and microRNAs (miRNAs) in 50 women suffering from PE in comparison with 50 healthy pregnant women.\n\n\nMethods\nExpressions of cytokines, specific transcription factors, and related miRNAs were measured by real‐time polymerase chain reaction (PCR). Enzyme‐linked immunosorbent assay (ELISA) was used to test the interleukin (IL)‐17, IL‐23, IL‐6, and IL‐10 and transforming growth factor β in serum and supernatant of peripheral blood mononuclear cells (PBMCs). The frequency of Th17 and Treg cells were determined by flow cytometry.\n\n\nResults\nPE patients exhibited a decreased number of Treg cells (p = 0.006), while Th17 cells were increased (\np = 0.004). Forkhead box P3 and \nIL‐10 mRNA expressions were reduced (\np = 0.0001 and 0.0028, respectively), while expressions of retinoic acid receptor‐related orphan nuclear receptor γt, IL‐17, IL‐23, and IL‐6 were enhanced (\np < 0.0001, 0.0018, 0.0014, and 0.027, respectively). ELISA results also showed increased levels of IL‐6, IL‐17, and IL‐23 (\np = 0.022, 0.0005, 0.0081, respectively), and decreased levels of IL‐10 in the supernatant of PBMCs of PE patients compared with control group (\np = 0.0011). There was significant upregulation of miR‐106b and miR‐326 (\np = 0.0048 and 0.028, respectively) in PE patients in comparison with the control group.\n\n\nConclusions\nThese findings suggest that imbalance of Th17/Treg cells, regulated possibly via microRNAs, may be involved in the pathogenesis of PE, emphasizing on the importance of these cells in feto‐maternal immune cross‐talk.\n" - Journal of Cellular Physiology, EarlyView.
  

  October 02, 2018   doi: 10.1002/jcp.27315   open full text
• Dual‐specificity phosphatase 6 deletion protects the colonic epithelium against inflammation and promotes both proliferation and tumorigenesis.
  Katia Beaudry, Marie‐Josée Langlois, Amélie Montagne, Sébastien Cagnol, Julie C. Carrier, Nathalie Rivard.
  Journal of Cellular Physiology. October 01, 2018
  --- - "\nAbstract\nThe Ras/mitogen‐activated protein kinase (MAPK) pathway controls fundamental cellular processes such as proliferation, differentiation, and apoptosis. The dual‐specificity phosphatase 6 (DUSP6) regulates cytoplasmic MAPK signaling by dephosphorylating and inactivating extracellular signal‐regulated kinase (ERK1/2) MAPK. To determine the role of DUSP6 in the maintenance of intestinal homeostasis, we characterized the intestinal epithelial phenotype of \nDusp6 knockout (KO) mice under normal, oncogenic, and proinflammatory conditions. Our results show that loss of Dusp6 increased crypt depth and epithelial cell proliferation without altering colonic architecture. Crypt regeneration capacity was also enhanced, as revealed by ex vivo \nDusp6 KO organoid cultures. Additionally, loss of Dusp6 induced goblet cell expansion without affecting enteroendocrine and absorptive cell differentiation. Our data also demonstrate that \nDusp6 KO mice were protected from acute dextran sulfate sodium‐induced colitis, as opposed to wild‐type mice. In addition, \nDusp6 gene deletion markedly enhanced tumor load in \nApc\nMin/+ mice. Decreased DUSP6 expression by RNA interference in HT29 colorectal cancer cells enhanced ERK1/2 activation levels and promoted both anchorage‐independent growth in soft agar as well as invasion through Matrigel. Finally, \nDUSP6 mRNA expression in human colorectal tumors was decreased in advanced stage tumors compared with paired normal tissues. These results demonstrate that DUSP6 phosphatase, by controlling ERK1/2 activation, regulates colonic inflammatory responses, and protects the intestinal epithelium against oncogenic stress." - Journal of Cellular Physiology, EarlyView.
  

  October 01, 2018   doi: 10.1002/jcp.27420   open full text
• Cover Image, Volume 233, Number 11, November 2018.
  Tingru Pan, Xueyuan Hu, Tianqi Liu, Zhe Xu, Na Wan, Yiming Zhang, Shu Li.
  Journal of Cellular Physiology. October 01, 2018
  --- - - Journal of Cellular Physiology, Volume 233, Issue 11, Page i-i, November 2018.
  

  October 01, 2018   doi: 10.1002/jcp.27284   open full text
• Table of Contents, Editor's Choice, Highlights.
  
  Journal of Cellular Physiology. October 01, 2018
  --- - - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8399-8407, November 2018.
  

  October 01, 2018   doi: 10.1002/jcp.27285   open full text
• Proinflammatory macrophages impair skeletal muscle differentiation in obesity through secretion of tumor necrosis factor‐α via sustained activation of p38 mitogen‐activated protein kinase.
  Xueqiang Wang, Daina Zhao, Yajuan Cui, Shemin Lu, Dan Gao, Jiankang Liu.
  Journal of Cellular Physiology. September 28, 2018
  --- - |2 Obesity is associated with skeletal muscle loss and impaired myogenesis. Increased infiltration of proinflammatory macrophages in skeletal muscle is noted in obesity and is associated with muscle insulin resistance. However, whether the infiltrated macrophages can contribute to obesity‐induced muscle loss is unclear. In this study, we investigate macrophage and muscle differentiation markers in the quadriceps (QC), gastrocnemius, tibia anterior, and soleus muscles from obese mice that were fed a high‐fat diet for 16 weeks. Then, we examined the effect and mediator of macrophage‐secreted factors on myoblast differentiation in vitro. We found markedly increased levels of proinflammatory macrophage markers (F4/80 and CD11c) in the QC muscle compared with the other three muscle groups. Consistent with the increased levels of proinflammatory macrophage infiltration, the QC muscle also showed a significant reduction in the expression of muscle differentiation makers MYOD1 and myosin heavy chain. In in vitro studies, treatment of C2C12 myoblasts with Raw 264.7 macrophage‐conditioned medium (CM) significantly promoted cell proliferation and inhibited myoblast differentiation. Neutralization of tumor necrosis factor α (TNF‐α) in Raw 264.7 macrophage CM reversed the reduction of myoblast differentiation. Finally, we found that both macrophage CM and TNF‐α induced sustained activation of p38 mitogen‐activated protein kinase (MAPK) in C2C12 myoblasts. Together, our findings suggest that the increased infiltration of proinflammatory macrophages could contribute toward obesity‐induced muscle loss by secreting inflammatory cytokine TNF‐α via the p38 MAPK signaling pathway. - Journal of Cellular Physiology, EarlyView.
  

  September 28, 2018   doi: 10.1002/jcp.27012   open full text
• Microbiota‐derived lipopolysaccharide retards chondrocyte hypertrophy in the growth plate through elevating Sox9 expression.
  Xin Cheng, Pei‐Zhi Li, Guang Wang, Yu Yan, Ke Li, Beate Brand‐Saberi, Xuesong Yang.
  Journal of Cellular Physiology. September 28, 2018
  --- - |2- Abstract Accumulating data show that the cytotoxicity of bacterial lipopolysaccharides (LPS) from microbiota or infection is associated with many disorders observed in the clinics. However, it is still obscure whether or not embryonic osteogenesis is affected by the LPS exposure during gestation. Using the early chicken embryo model, we could demonstrate that LPS exposure inhibits chondrogenesis of the 8‐day chicken embryos by Alcian Blue‐staining and osteogenesis of 17‐day by Alcian Blue and Alizarin Red staining. Further analysis of the growth plates showed that the length of the proliferating zone (PZ) increases whereas that of the hypertrophic zone (HZ) decreased following LPS exposure. However there is no significant change on cell proliferation in the growth plates. Immunofluorescent staining, western blot analysis, and quantitive polymerase chain reaction revealed that Sox9 and Col2a1 are highly expressed at the messenger RNA level and their protein products are also abundant. LPS exposure causes a downregulation of Runx2 and Col10a1 expression in 8‐day hindlimbs, and a suppression of Runx2, Col10a1, and Vegfa expression in 17‐day phalanges. Knocking down Sox9 in ATDC5 cells by small interfering RNA transfection lead to the expression reduction of Col2a1, Runx2, and Col10a1, implying the vital role of Sox9 in the process of LPS‐induced delay in the transition from proliferating chondrocytes to hypertrophic chondrocytes in the growth plate. In the presence of LPS, the antioxidant defense regulator nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) is highly expressed, and the activities of superoxide dismutase 1 (SOD1), SOD2, and glutaredoxin rise in 17‐day phalanges and ADTC5 cells. Simultaneously, an increase of intracellular ROS is observed. When Nrf2 expression was knocked down in ATDC5 cells, the expressions of Sox9, Col2a1, Runx2, Col10a1, and Vegfa were also going down as well. Taken together, our current data suggest that LPS exposure during gestation could restrict the chondrocytes conversion from proliferating to hypertrophic in the growth plate, in which LPS‐induced Sox9 plays a crucial role to trigger the cascade of downstream genes by excessive ROS production and Nrf2 elevation. - Journal of Cellular Physiology, EarlyView.
  

  September 28, 2018   doi: 10.1002/jcp.27025   open full text
• Curcumin attenuates proangiogenic and proinflammatory factors in human eutopic endometrial stromal cells through the NF‐κB signaling pathway.
  Indrajit Chowdhury, Saswati Banerjee, Adel Driss, Wei Xu, Sherifeh Mehrabi, Ceana Nezhat, Neil Sidell, Robert N. Taylor, Winston E. Thompson.
  Journal of Cellular Physiology. September 27, 2018
  --- - |2- Abstract Endometriosis is a chronic gynecological inflammatory disorder in which immune system dysregulation is thought to play a role in its initiation and progression. Due to altered sex steroid receptor concentrations and other signaling defects, eutopic endometriotic tissues have an attenuated response to progesterone. This progesterone‐resistance contributes to lesion survival, proliferation, pain, and infertility. The current agency‐approved hormonal therapies, including synthetic progestins, GnRH agonists, and danazol are often of limited efficacy and counterproductive to fertility and cause systemic side effects due to suppression of endogenous steroid hormone levels. In the current study, we examined the effects of curcumin (CUR, diferuloylmethane), which has long been used as an anti‐inflammatory folk medicine in Asian countries for this condition. The basal levels of proinflammatory and proangiogenic chemokines and cytokines expression were higher in primary cultures of stromal cells derived from eutopic endometrium of endometriosis (EESC) subjects compared with normal endometrial stromal cells (NESC). The treatment of EESC and NESC with CUR significantly and dose‐dependently reduced chemokine and cytokine secretion over the time course. Notably, CUR treatment significantly decreased phosphorylation of the IKKα/β, NF‐κB, STAT3, and JNK signaling pathways under these experimental conditions. Taken together, our findings suggest that CUR has therapeutic potential to abrogate aberrant activation of chemokines and cytokines, and IKKα/β, NF‐κB, STAT3, and JNK signaling pathways to reduce inflammation associated with endometriosis. - Journal of Cellular Physiology, EarlyView.
  

  September 27, 2018   doi: 10.1002/jcp.27360   open full text
• Blockade of NEAT1 represses inflammation response and lipid uptake via modulating miR‐342‐3p in human macrophages THP‐1 cells.
  Lei Wang, Jing‐Wen Xia, Zun‐Ping Ke, Bing‐Hong Zhang.
  Journal of Cellular Physiology. September 27, 2018
  --- - |2- Abstract Atherosclerosis has been recognized as a chronic inflammation process induced by lipid of the vessel wall. Oxidized low‐density lipoprotein (ox‐LDL) can drive atherosclerosis progression involving macrophages. Recently, long noncoding RNAs (lncRNAs) have been reported to play critical roles in atherosclerosis development. In our current study, we focused on the biological roles of lncRNA NEAT1 in atherosclerosis progress. Here, we found that ox‐LDL was able to trigger human macrophages THP‐1 cells, a human monocytic cell line, apoptosis in a dose‐dependent and time‐dependent course. In addition, we observed that NEAT1 was significantly increased in THP‐1 cells incubated with ox‐LDL and meanwhile miR‐342‐3p was greatly decreased. Then, NEAT1 was silenced by transfection of small interfering RNA (siRNA) of NEAT1 into THP‐1 cells. As exhibited, CD36, oil‐red staining levels, total cholesterol (TC), total cholesterol (TG) levels and THP‐1 cell apoptosis were obviously repressed by knockdown of NEAT1. Furthermore, inhibition of NEAT1 contributed to the repression of inflammation in vitro. Interleukin 6 (IL‐6), IL‐1β, cyclooxygenase‐2 (COX‐2) and tumour necrosis factor‐alpha (TNF‐α) protein levels were remarkably depressed by NEAT1 siRNA in THP‐1 cells. By using bioinformatics analysis, miR‐342‐3p was predicted as a downstream target of NEAT1 and the correlation between them was confirmed in our study. Moreover, overexpression of miR‐342‐3p could also greatly suppress inflammation response and lipid uptake in THP‐1 cells. Knockdown of NEAT1 and miR‐342‐3p mimics inhibited lipid uptake in THP‐1 cells. In conclusion, we implied that blockade of NEAT1 repressed inflammation response through modulating miR‐342‐3p in human macrophages THP‐1 cells and NEAT1 may offer a promising strategy to treat atherosclerotic cardiovascular diseases. - Journal of Cellular Physiology, EarlyView.
  

  September 27, 2018   doi: 10.1002/jcp.27340   open full text
• Notch‐1 signaling activation sustains overexpression of interleukin 33 in the epithelium of nasal polyps.
  G. Chiappara, S. Sciarrino, C. Di Sano, S. Gallina, R. Speciale, F. Lorusso, S. Di Vincenzo, C. D’Anna, A. Bruno, M. Gjomarkaj, E. Pace.
  Journal of Cellular Physiology. September 27, 2018
  --- - "\nAbstract\n\nBackground\nAlterations in the nasal epithelial barrier homeostasis and increased interleukin 33 (IL‐33) expression contribute to the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP).\n\n\nAims\nAs Notch‐1 signaling is crucial in repair processes of mucosa, the current study assessed Notch‐1/Jagged‐1 signaling and IL‐33 in the epithelium of nasal polyps biopsies from allergic (A‐CRSwNP; \nn = 9) and not allergic (NA‐CRSwNP; \nn = 9) subjects by immunohistochemistry. We also assessed, in a model of nasal epithelial cells, the effects of stimulation of Notch‐1 with Jagged‐1 on the expression of IL‐33 (by flow cytometry, immunofluorescence, and immunocytochemistry), Jagged‐1 (by flow cytometry), and p‐CREB transcription factor (by western blot analysis).\n\n\nResults\nEx vivo (a) in normal epithelium, the expression of Notch‐1 and IL‐33 were higher in NA‐CRSwNP than in A‐CRSwNP; (b) in metaplastic epithelium, the expression of Notch‐1, Jagged‐1, and IL‐33 were higher in NA‐CRSwNP than in A‐CRSwNP; (c) in hyperplastic epithelium, the expression of Notch‐1, Jagged‐1, and IL‐33 were higher in A‐CRSwNP than in NA‐CRSwNP; and (d) in basal epithelial cells, no differences were observed in the expression of Jagged‐1, IL‐33, and Notch‐1. The expression of Notch‐1 significantly correlated with the expression of IL‐33. In vitro, stimulation of Notch‐1 with Jagged‐1 induced the expression of (a) Jagged‐1; (b) IL‐33; and (c) p‐CREB transcription factor. The inhibitor of Notch‐1, DAPT, reduced all the effects of Jagged‐1 on nasal epithelial cells.\n\n\nConclusions\nThe data herein provided support, for the first time, a putative role of Notch‐1/Jagged‐1 signaling in the overexpression of IL‐33 in the epithelium of nasal polyps from patients with CRSwNP.\n" - Journal of Cellular Physiology, EarlyView.
  

  September 27, 2018   doi: 10.1002/jcp.27237   open full text
• Ewing‐like sarcoma: An emerging family of round cell sarcomas.
  Samuele Renzi, Nathaniel D. Anderson, Nicholas Light, Abha Gupta.
  Journal of Cellular Physiology. September 26, 2018
  --- - "\nAbstract\nEwing‐like sarcomas are an emerging subgroup of small round blue cell sarcomas that share various degrees of morphological, immunohistochemical, molecular, and clinical similarity with Ewing sarcoma. Despite these similarities, Ewing‐like sarcomas lack the pathognomonic molecular hallmark of Ewing sarcoma: A translocation between a gene of the RNA‐binding TET family (EWSR1 or \nFUS) with a gene of the ETS‐transcription family (\nFLI1, \nERG, \nETV1, \nETV4, or \nFEV). Recently, increased use of modern molecular methods based on next‐generation sequencing have enabled the identification of distinct subgroups within this previously uncharacterized group of Ewing‐like sarcomas based on the discovery of novel molecular driving events. The focus of this review is to provide an update on the main subcategories of Ewing‐like sarcomas discovered to date: \nCIC‐rearranged sarcomas, \nBCOR‐rearranged sarcomas, sarcomas with a rearrangement between \nEWSR1 and a non‐ETS family gene, and the substantial fraction of tumors which remain uncharacterized by molecular methods. There is increasing evidence that these tumors represent stand‐alone entities with unique characteristics rather than simply a subgroup of Ewing sarcoma; thus, the question of the best therapeutic approach for these often aggressive sarcomas remains of primary importance. Ultimately, large collaborative efforts will be necessary to better determine the characteristics of this rare, heterogeneous family of tumors." - Journal of Cellular Physiology, EarlyView.
  

  September 26, 2018   doi: 10.1002/jcp.27558   open full text
• Raloxifene attenuates oxidative stress and preserves mitochondrial function in astrocytic cells upon glucose deprivation.
  Diego J. Vesga‐Jiménez, Oscar Hidalgo‐Lanussa, Eliana Baez‐Jurado, Valentina Echeverria, Ghulam Md Ashraf, Amirhossein Sahebkar, George E. Barreto.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Oxidative stress and mitochondrial dysfunction induced by metabolic insults are both hallmarks of various neurological disorders, whereby neuronal cells are severely affected by decreased glucose supply to the brain. Likely injured, astrocytes are important for neuronal homeostasis and therapeutic strategies should be directed towards improving astrocytic functions to improve brain's outcome. In the present study, we aimed to assess the actions of raloxifene, a selective estrogen receptor modulator in astrocytic cells under glucose deprivation. Our findings indicated that pretreatment with 1 µM raloxifene results in an increase in cell viability and attenuated nuclei fragmentation. Raloxifene's actions also rely on the reduction of oxidative stress and preservation of mitochondrial function in glucose‐deprived astrocytic cells, suggesting the possible direct effects of this compound on mitochondria. In conclusion, our results demonstrate that raloxifene's protective actions might be mediated in part by astrocytes in the setting of a metabolic insult. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27481   open full text
• TGF‐β1‐PML SUMOylation‐peptidyl‐prolyl cis–trans isomerase NIMA‐interacting 1 (Pin1) form a positive feedback loop to regulate cardiac fibrosis.
  Di Wu, Di Huang, Liang‐Liang Li, Ping Ni, Xiu‐Xian Li, Bing Wang, Yan‐Na Han, Xiao‐Qi Shao, Dan Zhao, Wen‐Feng Chu, Bai‐Yan Li.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Transforming growth factor‐β (TGF‐β) signaling pathway is involved in fibrosis in most, if not all forms of cardiac diseases. Here, we evaluate a positive feedback signaling the loop of TGF‐β1/promyelocytic leukemia (PML) SUMOylation/Pin1 promoting the cardiac fibrosis. To test this hypothesis, the mice underwent transverse aortic constriction (3 weeks) were developed and the morphological evidence showed obvious interstitial fibrosis with TGF‐β1, Pin1 upregulation, and increase in PML SUMOylation. In neonatal mouse cardiac fibroblasts (NMCFs), we found that exogenous TGF‐β1 induced the upregulation of TGF‐β1 itself in a time‐ and dose‐dependent manner, and also triggered the PML SUMOylation and the formation of PML nuclear bodies (PML‐NBs), and consequently recruited Pin1 into nuclear to colocalize with PML. Pharmacological inhibition of TGF‐β signal or Pin1 with LY364947 (3 μM) or Juglone (3 μM), the TGF‐β1‐induced PML SUMOylation was reduced significantly with downregulation of the messenger RNA and protein for TGF‐β1 and Pin1. To verify the cellular function of PML by means of gain‐ or loss‐of‐function, the positive feedback signaling loop was enhanced or declined, meanwhile, TGF‐β‐Smad signaling pathway was activated or weakened, respectively. In summary, we uncovered a novel reciprocal loop of TGF‐β1/PML SUMOylation/Pin1 leading to myocardial fibrosis. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27357   open full text
• 2‐Methoxyestradiol attenuates chronic‐intermittent‐hypoxia‐induced pulmonary hypertension through regulating microRNA‐223.
  Shengyu Hao, Liyan Jiang, Cuiping Fu, Xu Wu, Zilong Liu, Jieqiong Song, Huan Lu, Xiaodan Wu, Shanqun Li.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Pulmonary hypertension (PH) is prevalent in patients with obstructive sleep apnea (OSA) syndrome, and coexistence of PH and OSA indicates a worse prognosis and higher mortality. Chronic intermittent hypoxia (CIH) is the key pathogenesis of OSA. Also, microRNA‐223 (miR‐223) plays a role in the regulation of CIH‐induced PH process. However, the detailed mechanism of CIH inducing PH is still unclear. This study aimed to investigate the pathological process of CIH associated PH and explore the potential therapeutic methods. In this study, adult Sprague–Dawley rats were exposed to CIH or normoxic (N) conditions with 2‐methoxyestradiol (2‐Me) or vehicle treatment for 6 weeks. The results showed that 2‐Me treatment reduced the progression of pulmonary angiogenesis in CIH rats, and alleviated proliferation, cellular migration, and reactive oxygen species formation was induced by CIH in pulmonary artery smooth muscle cells (PASMCs). CIH decreased the expression of miR‐223, whereas 2‐Me reversed the downregulation of miR‐223 both in vivo and in vitro. Furthermore, the antiangiogenic effect of 2‐Me observed in PASMCs was abrogated by miR‐223 inhibitor, while enhanced by miR‐223 mimic. These findings suggested that miR‐223 played an important role in the process of CIH inducing PH, and 2‐Me might reverse CIH‐induced PH via upregulating miR‐223. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27363   open full text
• Curcumin induces concentration‐dependent alterations in mitochondrial function through ROS in C2C12 mouse myoblasts.
  Tianzheng Yu, Jacob Dohl, Falicia Elenberg, Yifan Chen, Patricia Deuster.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Curcumin exhibits antioxidant properties in normal cells where the uptake is low, unlike in tumor cells where uptake is high and curcumin increases reactive oxygen species (ROS) production and cell death. Mitochondria are the main source and primary target of cellular ROS. We hypothesized that curcumin would regulate cellular redox status and mitochondrial function, depending on cell sensitivity and/or curcumin concentration in normal cells. We examined the differences between low and high concentrations of curcumin, with specific attention focused on ROS levels, mitochondrial function, and cell viability in mouse C2C12 myoblast under normal and simulated conditions of diabetes. Cells incubated with high concentrations of curcumin (10–50 μM) resulted in decreased cell viability and sustained robust increases in ROS levels. Mechanistic studies showed that increased ROS levels in cells incubated with 20 μM curcumin induced opening of mitochondrial permeability transition pores and subsequent release of cytochrome c, activation of caspases 9 and 3/7, and apoptotic cell death. Low concentrations of curcumin (1–5 μM) did not affect cell viability, but induced a mild increase in ROS levels, which peaked at 2 hr after the treatment. Incubation with 5 μM curcumin also induced ROS‐dependent increases in mitochondrial mass and membrane potential. Finally, pretreatment with 5 μM curcumin prevented high glucose‐induced oxidative cell injury. Our study suggests that mitochondria respond differentially depending on curcumin concentration‐dependent induction of ROS. The end result is either cell protection or death. Curcumin may be an effective therapeutic target for diabetes and other mitochondrial diseases when used in low concentrations. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27370   open full text
• Distinct Tie2 tyrosine phosphorylation sites dictate phenotypic switching in endothelial progenitor cells.
  Vahid Siavashi, Seyed Mahdi Nassiri, Reza Rahbarghazi, Zahra Mohseni, Ali Mohammad Sharifi.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Angiogenesis is a regulated process involving the proliferation, migration, and remodeling of different cell types particularly mature endothelial and their progenitor cells, nominated as endothelial progenitor cells (EPCs). Tie2/Tek is a tyrosine kinase receptor expressed by endothelial cells that induces signal transduction pathways involved in endothelial biology. To address the potential importance of the various tyrosine residues of Tie2 in EPC development, we generated a series of Tie2 tyrosine mutated (Y1106F, Y1100F, and Y1111F) EPCs and then assess the biological features of these cells. Clonogenic, tubulogenic, proliferative, migratory, and functional properties of these cells were analyzed. Next, GFP‐positive EPCs containing Tie2 tyrosine mutations were systemically transplanted into sublethaly irradiated mice to analyze the potency of these cells for marrow reconstitution. We found that mutation in the Tie2 tyrosine 1106 residue directed EPCs toward a mature endothelial phenotype, which was associated with augmented tubulogenic and migratory properties, and increased phosphorylation of the active site (tyrosine 992) as well as increased vascular perfusion in the in vivo Matrigel plug assay. Moreover, transplantation of 1106 Tie2 mutant EPCs failed to reconstitute the bone marrow after myeloablation, whereas transplantation of EPCs with the 1100 or 1111 Tie2 tyrosine mutation resulted in bone marrow engraftment, leading to improved survival of recipient mice. Our findings demonstrate that the tyrosine 1106 residue in Tie2 plays a key role to maintain the stemness features of EPCs. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27349   open full text
• HDAC8 regulates canonical Wnt pathway to promote differentiation in skeletal muscles.
  Luca Ferrari, Cinzia Bragato, Loredana Brioschi, Marco Spreafico, Simona Esposito, Alex Pezzotta, Fabrizio Pizzetti, Artal Moreno‐Fortuny, Gianfranco Bellipanni, Antonio Giordano, Paola Riva, Flavia Frabetti, Paola Viani, Giulio Cossu, Marina Mora, Anna Marozzi, Anna Pistocchi.
  Journal of Cellular Physiology. September 24, 2018
  --- - "\nAbstract\nHistone deacetylase 8 (HDAC8) is a class 1 histone deacetylase and a member of the cohesin complex. HDAC8 is expressed in smooth muscles, but its expression in skeletal muscle has not been described. We have shown for the first time that HDAC8 is expressed in human and zebrafish skeletal muscles. Using RD/12 and RD/18 rhabdomyosarcoma cells with low and high differentiation potency, respectively, we highlighted a specific correlation with \nHDAC8 expression and an advanced stage of muscle differentiation. We inhibited HDAC8 activity through a specific PCI‐34051 inhibitor in murine C2C12 myoblasts and zebrafish embryos, and we observed skeletal muscles differentiation impairment. We also found a positive regulation of the canonical Wnt signaling by HDAC8 that might explain muscle differentiation defects. These findings suggest a novel mechanism through which HDAC8 expression, in a specific time window of skeletal muscle development, positively regulates canonical Wnt pathway that is necessary for muscle differentiation." - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27341   open full text
• Long noncoding RNA SNHG12 promotes the progression of cervical cancer via modulating miR‐125b/STAT3 axis.
  Xue‐J. Jin, Xiang‐J. Chen, Zhi‐F. Zhang, Wen‐S. Hu, Rong‐Y. Ou, Shi Li, Ji‐S. Xue, Lu‐L. Chen, Yan Hu, Hua Zhu.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Increasing evidence showed that long noncoding RNAs (lncRNAs) played an important role in the occurrence and development of tumors. To date, lncRNA small nucleolar RNA host gene 12 (SNHG12) has revealed an oncogenic role in various tumors. However, the role of SNHG12 in cervical cancer is still unclear. Therefore, we focused on the biological function and molecular mechanism of SNHG12 in the tumorigenesis of cervical cancer. In this study, the expression of miR‐125b was observably downregulated in cervical cancer cells. Meanwhile, the expression of SNHG12 was obviously upregulated in cervical cancer cell lines (HeLa, SiHa, Caski, C4‐1, and C33A) compared with the immortalized cervical epithelial cells. The further assay showed that miR‐125b was a target of SNHG12 in cervical cancer. Moreover, a negative relationship between miR‐125b and SNHG12 was found in cervical cancer. In addition, SNHG12 inhibition restrained the proliferation, migration, and invasion of cervical cancer cells. Meanwhile, miR‐125b mimics repressed the expression of signal transducer and activator of transcription 3 (STAT3). The further assay showed that STAT3 was a target of miR‐125b in cervical cancer. In addition, sh‐STAT3 repressed the migration and invasion of cervical cancer cells. Furthermore, it showed that miR‐125b inhibitors reversed STAT3 expression restrained by the reduction of SNHG12 expression. In general, SNHG12 modulated STAT3 by sponging miR‐125b in cervical cancer and played an important role in the development of cervical cancer. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27403   open full text
• Autophagy participates in cyst breakdown and primordial folliculogenesis by reducing reactive oxygen species levels in perinatal mouse ovaries.
  Tu Zhihan, Mu Xinyi, Li Qingying, Gao Rufei, Zhang Yan, Chen Xuemei, Geng Yanqing, Wang Yingxiong, He Junlin.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract The reserve of primordial follicles, which serves all oocytes for the female reproductive lifespan, is established a few days after birth in mice. During this process, more than half of the oocytes are primarily eliminated by apoptosis. Autophagy, the conserved intracellular process maintaining cellular homeostasis, serves as a protective mechanism for oocyte survival. In the current study, we speculate a new role for autophagy during primordial folliculogenesis. Active autophagy was observed in perinatal ovaries from 16.5 days post coitus to 3 days post parturition. The inhibition of autophagy by 3‐methyladenine (3‐MA) increased the number of cyst oocytes and delayed follicle formation in vivo and in organ cultures. Furthermore, the reactive oxygen species (ROS) level was elevated in ovaries treated with 3‐MA, while N‐acetylcysteine, an oxidant, alleviated the inhibitory effect of 3‐MA on primordial folliculogenesis. Additionally, the expression of growth differentiation factor 9 and transforming growth factor β1, which regulates follicle activation, was decreased after 3‐MA treatment. These data suggest that the physiological level of autophagy in perinatal ovaries regulates germ cell cyst breakdown and primordial follicle assembly by ROS clearance and exerts extensive effects on further follicular development. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27367   open full text
• The pathogenesis of thyroid autoimmune diseases: New T lymphocytes – Cytokines circuits beyond the Th1−Th2 paradigm.
  Qian Li, Bin Wang, Kaida Mu, Jin‐An Zhang.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Autoimmune thyroid disease (AITD) is one of the most common organ‐specific autoimmune disorders. It mainly manifests as Hashimoto's thyroiditis (HT) and Graves’ disease (GD). HT is characteristic of hypothyroidism resulting from the destruction of the thyroid while GD is characteristic of hyperthyroidism due to excessive production of thyroid hormone induced by thyrotropin receptor‐specific stimulatory autoantibodies. T lymphocytes and their secretory cytokines play indispensable roles in modulating immune responses, but their roles are often complex and full of interactions among distinct components of the immune system. Dysfunction of these T cells or aberrant expressions of these cytokines can cause the breakdown of immune tolerance and result in aberrant immune responses during the development of AITDs. This review summarizes recently identified T subsets and related cytokines and their roles in the pathogenesis of AITDs with the hope to provide a better understanding of the precise roles of notably identified T subsets in AITDs and facilitate the discovery of functional molecules or novel immune therapeutic targets for AITDs. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27180   open full text
• Deglycosylation of epithelial cell adhesion molecule affects epithelial to mesenchymal transition in breast cancer cells.
  Xue Liu, Liu Yang, Dandan Zhang, Tingjiao Liu, Qiu Yan, Xuesong Yang.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract The transmembrane glycoprotein epithelial cell adhesion molecule (EpCAM) is overexpressed in most epithelial cancers including breast cancer, where it plays an important role in cancer progression. Previous study has demonstrated that knockdown of EpCAM inhibits breast cancer cell growth and metastasis via inhibition of the Ras/Raf/ERK signaling pathway and matrix metallopeptidase‐9 (MMP‐9). Although glycosylation is believed to be associated with the function of EpCAM, the contribution of N‐glycosylation to this function remains unclear. We constructed the N‐glycosylation mutation plasmid of EpCAM and used it to treat breast cancer cells. Loss of N‐glycosylation at all three sites EpCAM had no effect on its level of expression or membrane localization. However, mutation at glycosylation sites significantly reduced the ability of EpCAM to promote epithelial to mesenchymal transition in breast cancer. N‐glycosylation mutation of EpCAM led to decrease phosphorylation of Raf, ERK, and Akt, and inhibited the Ras/Raf/ERK and PI3K/Akt signaling pathways. Furthermore, we demonstrated that N‐glycosylation mutation of EpCAM‐mediated invasion and metastasis of breast carcinoma cells required the downregulation of MMP‐9 via inhibition of these two signaling pathways. Our results identified the characteristics and function of EpCAM glycosylation. These data could illuminate molecular regulation of EpCAM by glycosylation and promote our understanding of the application of glycosylated EpCAM as a target for breast cancer therapy. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27256   open full text
• PCSK9: A novel inflammation modulator in atherosclerosis?
  Zhi‐Han Tang, Tao‐Hua Li, Juan Peng, Jie Zheng, Ting‐Ting Li, Lu‐Shan Liu, Zhi‐Sheng Jiang, Xi‐Long Zheng.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Proprotein convertase subtilisin/kexin 9 (PCSK9) is the ninth member of the secretory serine protease family. It binds to low‐density lipoprotein receptor (LDLR) for endocytosis and lysosome degradation in the liver, resulting in an increasing in circulating LDL‐cholesterol (LDL‐c) level. Since a PCSK9 induced increase in plasma LDL‐c contributes to atherosclerosis, PCSK9 inhibition has become a new strategy in preventing and treating atherosclerosis. However, in addition to the effect of PCSK9 on elevating blood LDL‐c levels, accumulating evidence shows that PCSK9 plays an important role in inflammation, likely representing another major mechanism for PCSK9 to promote atherosclerosis. In this review, we discuss the association of PCSK9 and inflammation, and highlight the specific effects of PCSK9 on different vascular cellular components involved in the atherosclerotic inflammation. We also discuss the clinical evidence for the association between PCSK9 and inflammation in atherosclerotic cardiovascular disease. A better understanding of the direct association of PCSK9 with atherosclerotic inflammation might help establish a new role for PCSK9 in vascular biology and identify a novel molecular mechanism for PCSK9 therapy. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27254   open full text
• Angiotensin (1–7) inhibits arecoline‐induced migration and collagen synthesis in human oral myofibroblasts via inhibiting NLRP3 inflammasome activation.
  Yuehua You, Yun Huang, Dan Wang, Yang Li, Guozhen Wang, Siyi Jin, Xintao Zhu, Bin Wu, Xinya Du, Xu Li.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Arecoline induces oral submucous fibrosis (OSF) via promoting the reactive oxygen species (ROS). Angiotensin (1–7) (Ang‐(1–7)) protects against fibrosis by counteracting angiotensin II (Ang‐II) via the Mas receptor. However, the effects of Ang‐(1–7) on OSF remain unknown. NOD‐like receptors (NLRs) family pyrin domain containing 3 (NLRP3) inflammasome is identified as the novel mechanism of fibrosis. Whereas the effects of arecoline on NLRP3 inflammasome remain unclear. We aimed to explore the effect of Ang‐(1–7) on NLRP3 inflammasome in human oral myofibroblasts. In vivo, activation of NLRP3 inflammasomes with an increase of Ang‐II type 1 receptor (AT1R) protein level and ROS production in human oral fibrosis tissues. Ang‐(1–7) improved arecoline‐induced rats OSF, reduced protein levels of NADPH oxidase 4 (NOX4) and the NLRP3 inflammasome. In vitro, arecoline increased ROS along with upregulation of the angiotensin‐converting enzyme (ACE)/Ang‐II/AT1R axis and NLRP3 inflammasome/interleukin‐1β axis in human oral myofibroblasts, which were reduced by NOX4 inhibitor VAS2870, ROS scavenger N‐acetylcysteine, and NOX4 small interfering RNA (siRNA). Furthermore, arecoline induced collagen synthesis or migration via the Smad or RhoA‐ROCK pathway respectively, which could be inhibited by NLRP3 siRNA or caspase‐1 blocker VX‐765. Ang‐(1–7) shifted the balance of RAS toward the ACE2/Ang‐(1–7)/Mas axis, inhibited arecoline‐induced ROS and NLRP3 inflammasome activation, leading to attenuation of migration or collagen synthesis. In summary, Ang‐(1–7) attenuates arecoline‐induced migration and collagen synthesis via inhibiting NLRP3 inflammasome in human oral myofibroblasts. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27267   open full text
• The miR‐15a/16 gene cluster in human cancer: A systematic review.
  Ting Liu, Zhenru Xu, Daming Ou, Jing Liu, Ji Zhang.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract MicroRNAs (miRNAs) are an important class of endogenous small noncoding single‐stranded RNAs that suppress the expression of their target genes through messenger RNA (mRNA) degradation to inhibit transcription and translation. MiRNAs play a crucial regulatory role in many biological processes including proliferation, metabolism, and cellular malignancy. miR‐15a/16 is an important tumor suppressor gene cluster with a variety of factors that regulate its transcriptional activity. It has been discovered that a relative reduction of miR‐15a/16 expression in various cancers is closely related to the occurrence and progression of tumors. miR‐15a/16 takes part in a wide array of biological processes including tumor cell proliferation, apoptosis, invasion, and chemoresistance by binding to the 3′‐untranslated region of its target gene's mRNA. In this review, we will examine the complex regulatory network of miR‐15a/16 gene expression and its biological functions in human cancers to further elucidate the molecular mechanisms of its antitumor effects. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27342   open full text
• Potentiating apoptosis and modulation of p53, Bcl2, and Bax by a novel chrysin ruthenium complex for effective chemotherapeutic efficacy against breast cancer.
  Souvik Roy, Anweshan Sil, Tania Chakraborty.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Breast cancer is the most frequent cause of cancer in women. In the current study, transition metal ruthenium was complexed with flavonoid chrysin to evaluate the chemotherapeutic potential of this compound in Michigan Cancer Foundation‐7 (MCF‐7) human mammary cancer cell line and 7,12‐dimethylbenz(α)anthracene‐induced mammary cancer in female Sprague–Dawley rats. The characterizations of the complex were accomplished through UV–visible, NMR, IR, Mass spectra, and XRD techniques and antioxidant activity was assessed by DPPH, FRAP, and ABTS methods. In vitro studies included cell viability, cell cycle analysis, DNA fragmentation, and marker analysis by western blot analysis and found that complex treatment suppressed cell growth‐induced cell cycle arrest and enhanced the induction of apoptosis in cancer cells. Moreover, complex treatment modulated signaling pathways including mTOR, VEGF, and p53 in the MCF‐7 cells. Acute and subacute toxicity was performed in rats to determine the therapeutic doses. Breast cancer in rats was initiated by the administration of 7,12‐dimethylbenz(α)anthracene (0.5 mg/100 g body weight) via single tail vein injection. The histopathological analysis after 24 weeks of carcinogenesis study depicted substantial repair of hyperplastic lesions. Immunohistochemical analysis revealed upregulation of Bax and p53 and downregulation of Bcl2 proteins and TUNEL assay showed an increase in apoptotic index in ruthenium–chrysin‐treated groups as compared to the carcinogen control. Our findings from the in vitro and in vivo study support the continued investigation of ruthenium–chrysin complex possesses a potential chemotherapeutic activity against breast cancer and was efficient in reducing hyperplastic lesions in the mammary tissues of rats by inducing apoptosis. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27287   open full text
• 1,7‐Bis(4‐hydroxyphenyl)‐1,4‐heptadien‐3‐one induces lung cancer cell apoptosis via the PI3K/Akt and ERK1/2 pathways.
  Jiangjiang Fan, Mingsheng Wu, Jian Wang, Dongmei Ren, Jian Zhao, Guotao Yang.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract 1,7‐Bis(4‐hydroxyphenyl)‐1,4‐heptadien‐3‐one (EB30) is a diarylheptanoid‐like compound isolated from Viscum coloratum. This curcumin analog exhibits significant cytotoxic activity against HeLa, SGC‐7901, and MCF‐7 cells. However, little is known about the anticancer effects and mechanisms of EB30 in human lung cancer. The current study reports that EB30 significantly reduced the cell viability of A549 and NCI‐H292 human lung cancer cells. Further examination revealed that EB30 not only induced cell cycle arrest and promoted the generation of reactive oxygen species (ROS) but also induced cell apoptosis through the intrinsic and extrinsic signaling pathways. Furthermore, EB30 upregulated the expression levels of p‐ERK1/2 and p‐P90RSK, whereas downregulating the phosphorylation of Akt and P70RSK. Cell viability was further inhibited by the combination of EB30 with LY294002 (a specific PI3K inhibitor) or U0126 (a MEK inhibitor). The current study indicates that EB30 is a potential anticancer agent that induces cell apoptosis via suppression of the PI3K/Akt pathway and activation of the ERK1/2 pathway. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27364   open full text
• Tannic acid protects against experimental acute lung injury through downregulation of TLR4 and MAPK.
  Ayyanar Sivanantham, Dhamotharan Pattarayan, Ramalingam Bethunaickan, Amrita Kar, Santanu Kar Mahapatra, Rajesh K. Thimmulappa, Rajaguru Palanichamy, Subbiah Rajasekaran.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2 Abstract Acute lung injury (ALI) and its severe form acute respiratory distress syndrome (ARDS) remain a major cause of morbidity and mortality in critically ill patients, and no specific therapies are still available to control the mortality rate. Thus, we explored the preventive and therapeutic effects of tannic acid (TA), a natural polyphenol in the context of ALI. We used in vivo and in vitro models, respectively, using lipopolysaccharide (LPS) to induce ALI in mice and exposing J774 and BEAS‐2B cells to LPS. In both preventive and therapeutic approaches, TA attenuated LPS‐induced histopathological alterations, lipid peroxidation, lung permeability, infiltration of inflammatory cells, and the expression of proinflammatory mediators. In addition, in‐vitro study showed that TA treatment could reduce the expression of proinflammatory mediators. Further studies revealed that TA‐dampened inflammatory responses by downregulating the LPS‐induced toll‐like receptor 4 (TLR4) expression and inhibiting extracellular‐signal‐regulated kinase (ERK)1/2 and p38 mitogen‐activated protein kinase (MAPK) activation. Furthermore, cells treated with the inhibitors of ERK1/2 (PD98059) and p38 (SB203580) mitigated the expression of cytokines induced by LPS, thus suggesting that ERK1/2 and p38 activity are required for the inflammatory response. In conclusion, TA could attenuate LPS‐induced inflammation and may be a potential therapeutic agent for ALI‐associated inflammation in clinical settings. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27383   open full text
• Prognostic value of aberrantly expressed methylation gene profiles in lung squamous cell carcinoma: A study based on The Cancer Genome Atlas.
  Chundi Gao, Jing Zhuang, Chao Zhou, Ke Ma, Minzhang Zhao, Cun Liu, Lijuan Liu, Huayao Li, Fubin Feng, Changgang Sun.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Currently, research on genome‐scale epigenetic modifications for studying the pathogenesis of lung cancer is lacking. Aberrant DNA methylation, as the most common and important modification in epigenetics, is an important means of regulating genomic function and can be used as a biomarker for the diagnosis and prognosis of lung squamous cell carcinoma (LUSC). In this paper, methylation information and gene expression data from patients with LUSC were extracted from the TCGA database. Univariate and multivariate COX analyses were used to screen abnormally methylated genes related to the prognosis of LUSC. The relationship between key DNA methylation sites and the transcriptional expression of LUSC‐related genes was explored. A prognostic risk model constructed by four abnormally methylated genes (VAX1, CH25H, AdCyAP1, and Irx1) was used to predict the prognosis of LUSC patients. Also, the methylation levels of the key gene IRX1 are significantly correlated with the prognosis and correlated with the methylation of the site cg09232937 and cg10530883. This study is based on high‐throughput data mining and provides an effective bioinformatics basis for further understanding the pathogenesis and prognosis of LUSC, which has important theoretical significance for follow‐up studies on LUSC. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27389   open full text
• Dynamics changes in the transcription factors during early human embryonic development.
  Rasoul Godini, Hossein Fallahi.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Development of an embryo from a single cell, zygote, to multicellular morulae requires activation of hundreds of genes that were mostly inactivated before fertilization. Inevitably, transcription factors (TFs) would be involved in modulating the drastic changes in gene expression pattern observed at all preimplantation stages. Despite many ongoing efforts to uncover the role of TFs at the early stages of embryogenesis, still many unanswered questions remained that need to be explored. This could be done by studying the expression pattern of multiple genes obtained by high‐throughput techniques. In the current study, we have identified a set of TFs that are involved in the progression of the zygote to blastocyst. Global gene expression patterns of consecutive stages were compared and differences documented. Expectedly, at the early stages of development, only a few sets of TFs differentially expressed while at the later stages hundreds of TFs appear to be upregulated. Interestingly, the expression levels of many TFs show an oscillation pattern during development indicating a need for their precise expression. A significant shift in gene expression was observed during the transition from four‐ to eight‐cell stages, an indication of zygote genome activation. Additionally, we have found 11 TFs that were common in all stages including ATF3, EN1, IFI16, IKZF3, KLF3, NPAS3, NR2F2, RUNX1, SOX2, ZBTB20, and ZSCAN4. However, their expression patterns did not follow similar trends in the steps studied. Besides, our findings showed that both upregulation and active downregulation of the TFs expression is required for successful embryogenesis. Furthermore, our detailed network analysis identified the hub TFs for each transition. We found that HNF4A, FOXA2, and EP300 are the three most important elements for the first division of zygote. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27386   open full text
• USP49 inhibits ischemia–reperfusion‐induced cell viability suppression and apoptosis in human AC16 cardiomyocytes through DUSP1–JNK1/2 signaling.
  Wei Zhang, Yangyang Zhang, Hengbing Zhang, Qian Zhao, Zheng Liu, Yawei Xu.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Dual‐specificity protein phosphatases (DUSP) also known as mitogen‐activated protein kinase (MAPK) phosphatases (MKPs) can dephosphorylate MAPKs, including extracellular signal‐regulated kinase, c‐Jun N‐terminal kinase (JNK), and p38. DUSP1‐mediated JNK dephosphorylation has been found to play an antiapoptotic role against cardiac ischemia–reperfusion (I/R) injury. However, the regulation of DUSP1–JNK pathway remains unclear. In the current study, ubiquitin‐specific peptidase 49 (USP49) expression in human AC16 cardiomyocytes following I/R injury was measured by real‐time polymerase chain reaction and western blot analysis. Cell viability, apoptosis, the Bax, Bcl‐2, and DUSP1 expression, and the activity of MAPKs in AC16 cardiomyocytes following indicated treatment was measured by CCK‐8, flow cytometry, and western blot analysis. The direct interaction between USP49 and DUSP1 was measured by coimmunoprecipitation and ubiquitination analysis. The effect of USP49 on apoptosis and JNK activity in rat cardiomyocytes following I/R injury was also measured by TUNEL and western blot analysis. Here, we found that USP49 expression was time‐dependently increased in AC16 cardiomyocytes following I/R. I/R‐induced cell apoptosis and JNK1/2 activation both in in vivo and in vitro reversed by USP49 overexpression in AC16 cardiomyocytes. Inhibiting JNK1/2 activation significantly inhibited USP49 knockdown‐induced the cell viability inhibition, apoptosis and the JNK1/2 activation in AC16 cardiomyocytes. Moreover, USP49 positively regulated DUSP1 expression through deubiquitinating DUSP1. Overall, our findings establish USP49 as a novel regulator of DUSP1–JNK1/2 signaling pathway with a protective role in cardiac I/R injury. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27390   open full text
• Long noncoding RNA NEAT1 modulates cell proliferation and apoptosis by regulating miR‐23a‐3p/SMC1A in acute myeloid leukemia.
  Chen Zhao, Shanshan Wang, Yang Zhao, Feng Du, Weiyao Wang, Peng Lv, Ling Qi.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract The aim of this study was to determine the function of the NEAT1/miR‐23a‐3p/SMC1A axis in cell proliferation and apoptosis in acute myeloid leukemia (AML). Microarray analysis was used to screen differentially expressed lncRNAs/miRNAs/mRNAs in primary AML cells. The expression of nuclear paraspeckle assembly transcript 1 (NEAT1), miR‐23a‐3p, and structural maintenance of chromosome 1 alpha (SMC1A) in primary AML cells and THP‐1 cells were measured by quantitative real‐time polymerase chain reaction (qRT‐PCR). A Cell Counting Kit‐8 (CCK‐8) assay was used to analyze proliferation. Cell cycle progression and apoptosis were examined by flow cytometry. RNA immunoprecipitation (RIP) and dual‐luciferase assays were performed to determine the correlation between miR‐23a‐3p and NEAT1 or SMC1A. The qRT‐PCR illustrated that NEAT1 and SMC1A expression was decreased but that miR‐23a‐3p expression was increased in primary AML cells and THP‐1 cells compared with that in normal cells. The RIP assay and dual‐luciferase assay revealed the targeting relationship between miR‐23a‐3p and NEAT1 or SMC1A. The CCK‐8 assay showed that the overexpression of NEAT1 and SMC1A or repression of miR‐23a‐3p inhibited cell proliferation. Flow cytometry showed that the upregulation of NEAT1 and SMC1A or repression of miR‐23a‐3p promoted apoptosis and affected the cell cycle. NEAT1 repressed the expression of miR‐23a‐3p, and therefore promoted SMC1A, which in turn suppressed myeloid leukemia cell proliferation and enhanced apoptosis. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27393   open full text
• Basolateral presence of the proinflammatory cytokine tumor necrosis factor ‐α and secretions from adipocytes and macrophages reduce intestinal sugar transport.
  Rosa Castilla‐Madrigal, Eva Gil‐Iturbe, Neira Sáinz, María J. Moreno‐Aliaga, María Pilar Lostao.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- We have previously demonstrated in Caco‐2 cells that tumor necrosis factor‐α (TNF‐α) inhibits sugar uptake, acting from the apical membrane, by decreasing the expression of the Na+‐glucose cotransporter SGLT1 in the brush border membrane. The goal was to investigate the hypothesis that TNF‐α from abdominal adipose tissue (adipocytes and macrophages) would decrease sugar and amino acid transport acting from the basolateral membrane of the enterocytes. TNF‐α placed in the basal compartment of Caco‐2 cells decreased α‐methyl‐ d‐glucose (αMG) and glutamine uptake. The apical medium derived from these Caco‐2 cells apically placed in another set of cells, also reduced sugar and glutamine transport. Reverse‐transcription polymerase chain reaction analysis demonstrated upregulation of TNF‐α, IL‐1β, and MCP1 expression in Caco‐2 cells exposed to basal TNF‐α. Similarly, αMG uptake was inhibited after Caco‐2 cells were incubated, in the basal compartment, with medium from visceral human mesenchymal stem cells‐derived adipocytes of overweight individuals. The apical medium collected from those Caco‐2 cells, and placed in the upper side of other set of cells, also decreased sugar uptake. Basal presence of medium derived from lipopolysaccharide‐activated macrophages and nonactivated macrophages decreased αMG uptake as well. Diet‐induced obese mice showed an increase in the visceral adipose tissue surrounding the intestine. In this physiological condition, there was a reduction on αMG uptake in jejunal everted rings. Altogether, these results suggest that basolateral TNF‐α, which can be produced by adipocytes and macrophages during obesity, would be able to activate TNF‐α and other proinflammatory proteins expression in the small intestine and diminish intestinal sugar and amino acids transport. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27216   open full text
• Trastuzumab‐monomethyl auristatin E conjugate exhibits potent cytotoxic activity in vitro against HER2‐positive human breast cancer.
  Meghdad Abdollahpour‐Alitappeh, Majid Lotfinia, Nader Bagheri, Koushan Sineh Sepehr, Mahdi Habibi‐Anbouhi, Farzad Kobarfard, Saeed Balalaie, Alireza Foroumadi, Ghasem Abbaszadeh‐Goudarzi, Kazem Abbaszadeh‐Goudarzi, Mohsen Abolhassani.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2- Abstract Targeted therapy using specific monoclonal antibodies (mAbs) conjugated to chemotherapeutic agents or toxins has become one of the top priorities in cancer therapy. Antibody–drug conjugates (ADCs) are emerging as a promising strategy for cancer‐targeted therapy. In this study, trastuzumab, a humanized monoclonal anti‐HER2 antibody, was reduced by dithiothreitol and conjugated to the microtubule‐disrupting agent monomethyl auristatin E (MMAE) through a valine‐citrulline peptide linker (trastuzumab‐MC‐Val‐Cit‐PABC‐MMAE [trastuzumab‐vcMMAE]). After conjugation, ADCs were characterized by using UV–vis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE), and flow cytometry. The antitumor activity of the ADC was evaluated in breast cancer cells in vitro. In addition, ADCs were further characterized using purification by the protein A chromatography, followed by assessment using apoptosis and MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide) assays. Hydrophobic interaction chromatography was used to determine drug‐to‐antibody ratio species of ADCs produced. Our finding showed that approximately 5.12 drug molecules were conjugated to each mAb. H2L2, H2L, HL, H2, H, and L forms of ADCs were detected in nonreducing SDS‐PAGE. The binding of trastuzumab‐vcMMAE to HER2‐positive cells was comparable with that of the parental mAb. The MTT assay showed that our ADCs induced significant cell death in HER2‐positive cells, but not in HER2‐negative cells. The ADCs produced was a mixture of species, unconjugated trastuzumab (14.147%), as well as trastuzumab conjugated with two (44.868%), four (16.886%), six (13.238%), and eight (10.861%) molecules of MMAE. These results indicated that MMAE‐conjugated trastuzumab significantly increases the cytotoxic activity of trastuzumab, demonstrating high affinity, specificity, and antitumor activity in vitro. Trastuzumab‐vcMMAE is an effective and selective agent for the treatment of HER2‐positive breast tumors. - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27085   open full text
• Gene expression of TWIST1 and ZBTB16 is regulated by methylation modifications during the osteoblastic differentiation of mesenchymal stem cells.
  Faroogh Marofi, Ghasem Vahedi, Saeed Solali, Mohammadreza Alivand, Sadegh Salarinasab, Milad Zadi Heydarabad, Majid Farshdousti Hagh.
  Journal of Cellular Physiology. September 24, 2018
  --- - |2 Abstract Background Osteoblastic differentiation of mesenchymal stem cells (MSCs) is the principal stage during the restoration and regeneration of bone tissue. Epigenetic modifications such as DNA methylation play a key role in the differentiation process of stem cells. In this study, the methylation status of the promoter region of ZBTB16 and Twist1 genes and their role in controlling osteoblastic differentiation in MSCs was investigated during the osteoblastic differentiation of MSCs. Methods The MSCs were cultured under standard conditions and differentiated into the osteoblasts. We had three treatment groups including 5‐azacytidine (methylation inhibitor), metformin (Twist‐inhibitor), and procaine (Wnt/β‐catenin inhibitor) and a non‐treated group (control). Methylation level of DNA in the promoter regions was monitored by methylation specific‐quantitative polymerase chain reaction (PCR). Also, the mRNA levels of key genes in osteoblastic differentiation were measured using real‐time PCR. Results ZBTB16 gene expression was upregulated, and promoter methylation was decreased. For Twist1 messenger RNA (mRNA) level decreased and promoter methylation increased during osteoblastic differentiation of MSCs. 5‐Azacytidine caused a significant reduction in methylation and increased the mRNA expression of ZBTB16 and Twist1. Metformin repressed the Twist1 expression, and therefore osteoblastic differentiation was increased. On the opposite side, procaine could block the WNT/β‐catenin signaling pathway, as a consequence the gene expression of key genes involved in osteoblastic differentiation was declined. Conclusion We found that methylation of DNA in the promoter region of ZBTB16 and Twist1 genes might be one of the main mechanisms that controlling the gene expression during osteoblastic differentiation of MSCs. Also, we could find an association between regulation of Twist1 and ZBTB16 genes and osteoblastic differentiation in MSCs by showing the relation between their expression and some key genes involved in osteoblastic differentiation. In addition, we found a connection between the Twist1 expression level and osteoblastic differentiation by using a Twist‐inhibitor (metformin). - Journal of Cellular Physiology, EarlyView.
  

  September 24, 2018   doi: 10.1002/jcp.27352   open full text
• Cholesterol burden in the liver induces mitochondrial dynamic changes and resistance to apoptosis.
  Mayra Domínguez‐Pérez, Arturo Simoni‐Nieves, Patricia Rosales, Natalia Nuño‐Lámbarri, Mónica Rosas‐Lemus, Verónica Souza, Roxana U. Miranda, Leticia Bucio, Salvador Uribe Carvajal, Jens U. Marquardt, Daekwan Seo, Luis E. Gomez‐Quiroz, María Concepción Gutiérrez‐Ruiz.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Non‐alcoholic fatty liver disease (NAFLD) encompasses a broad spectrum of histopathological changes ranging from non‐inflammatory intracellular fat deposition to non‐alcoholic steatohepatitis (NASH), which may progress into hepatic fibrosis, cirrhosis, or hepatocellular carcinoma. Recent data suggest that impaired hepatic cholesterol homeostasis and its accumulation are relevant to the pathogenesis of NAFLD/NASH. Despite a vital physiological function of cholesterol, mitochondrial dysfunction is an important consequence of dietary‐induced hypercholesterolemia and was, subsequently, linked to many pathophysiological conditions. The aim in the current study was to evaluate the morphological and molecular changes of cholesterol overload in mouse liver and particularly, in mitochondria, induced by a high‐cholesterol (HC) diet for one month. Histopathological studies revealed microvesicular hepatic steatosis and significantly elevated levels of liver cholesterol and triglycerides leading to impaired liver synthesis. Further, high levels of oxidative stress could be determined in liver tissue as well as primary hepatocyte culture. Transcriptomic changes induced by the HC diet involved disruption in key pathways related to cell death and oxidative stress as well as upregulation of genes related to glutathione homeostasis. Impaired liver function could be associated with a decrease in mitochondrial membrane potential and ATP content and significant alterations in mitochondrial dynamics. We demonstrate that cholesterol overload in the liver leads to mitochondrial changes which may render damaged hepatocytes proliferative and resistant to cell death whereby perpetuating liver damage. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27474   open full text
• Upregulation of lncRNA DGCR5 correlates with better prognosis and inhibits bladder cancer progression via transcriptionally facilitating P21 expression.
  Chen Fang, Wei He, Tianyuan Xu, Jun Dai, Le Xu, Fukang Sun.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Mounting studies show that long noncoding RNAs (lncRNAs) could affect human cancer progression, including bladder cancer (BCa). LncRNA DiGeorge syndrome critical region gene 5 (DGCR5) has been proven to be involved in lung cancer, pancreatic ductal adenocarcinoma, and hepatocellular carcinoma. However, the function of DGCR5 in BCa remains largely unknown. Here, we found that DGCR5 expression was significantly downregulated in BCa tissues compared with adjacent normal tissues. Higher expression of DGCR5 predicted higher survival rate in BCa patients. Functional experiments indicated that DGCR5 overexpression markedly inhibited that proliferation, colony formation, and cell‐cycle progression in BCa cells. Furthermore, ectopic expression of DGCR5 led to decreased BCa cell migration, invasion, and epithelial–mesenchymal transition while promoting apoptosis. In vivo xenograft assay also illustrated that DGCR5 overexpression inhibited BCa growth. In the mechanism, we found that DGCR5 interacted with AT‐rich interaction domain 1A (ARID1A), a chromatin remodeling protein, to promote P21 transcription. Knockdown of P21 could significantly rescue the suppressed proliferation, migration, and invasion of BCa cells by DGCR5 overexpression. In summary, our study demonstrated that DGCR5 transcriptionally promotes P21 expression to suppress BCa progression. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27356   open full text
• Hypoxia induces ZEB2 in podocytes: Implications in the pathogenesis of proteinuria.
  Krishnamurthy Nakuluri, Dhanunjay Mukhi, Rajkishor Nishad, Moin A. Saleem, Sathish Kumar Mungamuri, Ram K. Menon, Anil Kumar Pasupulati.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract The glomerular filtration barrier (GFB) plays a critical role in ensuing protein free urine. The integrity of the GFB is compromised during hypoxia that prevails during extreme physiological conditions. However, the mechanism by which glomerular permselectivity is compromised during hypoxia remains enigmatic. Rats exposed to hypoxia showed a decreased glomerular filtration rate, podocyte foot‐processes effacement, and proteinuria. Accumulation of hypoxia‐inducible factor‐1α (HIF1α) in podocytes resulted in elevated expression of zinc finger E‐box binding homeobox 2 (ZEB2) and decreased expression of E‐ and P‐cadherin. We also demonstrated that HIF1α binds to hypoxia response element localized in the ZEB2 promoter. Furthermore, HIF1α also induced the expression of ZEB2‐natural antisense transcript, which is known to increase the efficiency of ZEB2 translation. Ectopic expression of ZEB2 induced loss of E‐ and P‐cadherin and is associated with enhanced motility of podocytes during hypoxic conditions. ZEB2 knockdown abrogated hypoxia‐induced decrease in podocyte permselectivity. This study suggests that hypoxia leads to activation of HIF1α–ZEB2 axis, resulting in podocyte injury and poor renal outcome. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27387   open full text
• Genetics and rheumatoid arthritis susceptibility in Iran.
  Shahla Korani, Mitra Korani, Alexandra E. Butler, Amirhossein Sahebkar.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Rheumatoid arthritis (RA) is an autoimmune disorder with a number of risk factors, including both genetic and environmental. A number of RA risk associated genomic loci has been identified. In this review, we summarize the association of genetic factors with RA reported in population studies in Iran. No significant association was found between the majority of genetic factors identified in other populations and risk for RA in the Iranian subjects. This conflicting result could be due to the ethnic differences and diversity that are present in Iran. We conclude that there is a need to investigate larger groups of Iranian subjects, encompassing different regions of Iran, to either prove or refute these initial findings. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27379   open full text
• Melatonin and cancer: From the promotion of genomic stability to use in cancer treatment.
  Bagher Farhood, Nasser Hashemi Goradel, Keywan Mortezaee, Neda Khanlarkhani, Masoud Najafi, Amirhossein Sahebkar.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Cancer remains among the most challenging human diseases. Several lines of evidence suggest that carcinogenesis is a complex process that is initiated by DNA damage. Exposure to clastogenic agents such as heavy metals, ionizing radiation (IR), and chemotherapy drugs may cause chronic mutations in the genomic material, leading to a phenomenon named genomic instability. Evidence suggests that genomic instability is responsible for cancer incidence after exposure to carcinogenic agents, and increases the risk of secondary cancers following treatment with radiotherapy or chemotherapy. Melatonin as the main product of the pineal gland is a promising hormone for preventing cancer and improving cancer treatment. Melatonin can directly neutralize toxic free radicals more efficiently compared with other classical antioxidants. In addition, melatonin is able to regulate the reduction/oxidation (redox) system in stress conditions. Through regulation of mitochondrial nction and inhibition of pro‐oxidant enzymes, melatonin suppresses chronic oxidative stress. Moreover, melatonin potently stimulates DNA damage responses that increase the tolerance of normal tissues to toxic effect of IR and may reduce the risk of genomic instability in patients who undergo radiotherapy. Through these mechanisms, melatonin attenuates several side effects of radiotherapy and chemotherapy. Interestingly, melatonin has shown some synergistic properties with IR and chemotherapy, which is distinct from classical antioxidants that are mainly used for the alleviation of adverse events of radiotherapy and chemotherapy. In this review, we describe the anticarcinogenic effects of melatonin and also its possible application in clinical oncology. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27391   open full text
• Identification of a novel cell cycle‐related gene signature predicting survival in patients with gastric cancer.
  Lan Zhao, Longyang Jiang, Linxiu He, Qian Wei, Jia Bi, Yan Wang, Lifeng Yu, Miao He, Lin Zhao, Minjie Wei.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Gastric cancer (GC) is one of the most fatal cancers in the world. Thousands of biomarkers have been explored that might be related to survival and prognosis via database mining. However, the prediction effect of single gene biomarkers is not specific enough. Increasing evidence suggests that gene signatures are emerging as a possible better alternative. We aimed to develop a novel gene signature to improve the prognosis prediction of GC. Using the messenger RNA (mRNA)‐mining approach, we performed mRNA expression profiling in a large GC cohort (n = 375) from The Cancer Genome Atlas (TCGA) database. Gene Set Enrichment Analysis (GSEA) was performed, and we recovered genes related to the G2/M checkpoint, which we identified with a Cox proportional regression model. We identified a set of five genes (MARCKS, CCNF, MAPK14, INCENP, and CHAF1A), which were significantly associated with overall survival (OS) in the test series. Based on this five‐gene signature, the test series patients could be classified into high‐risk or low‐risk subgroups. Multivariate Cox regression analysis indicated that the prognostic power of this five‐gene signature was independent of clinical features. In conclusion, we developed a five‐gene signature related to the cell cycle that can predict survival for GC. Our findings provide novel insight that is useful for understanding cell cycle mechanisms and for identifying patients with GC with poor prognoses. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27365   open full text
• A genetic variant in CDKN2A/2B locus was associated with poor prognosis in patients with esophageal squamous cell carcinoma.
  Niloofar Ghobadi, Mehrane Mehramiz, Soodabeh ShahidSales, Arezou Rezaei Brojerdi, Kazem Anvari, Majid Khazaei, Majid Rezayi, Mohammad Sadegh Khorrami, Mona Joudi‐Mashhad, Hassan Ramshini, Saeideh Ahmadi‐Simab, Ali Moradi, Seyed Mahdi Hassanian, Majid Ghayour‐Mobarhan, Mohammad Taher Boroushaki, Gordon A. Ferns, Amir Avan.
  Journal of Cellular Physiology. September 21, 2018
  --- - "\nAbstract\nEsophageal squamous cell carcinoma (ESCC) is among the leading causes of cancer related death. Despite of extensive efforts in identifying valid cancer prognostic biomarkers, only a very small number of markers have been identified. Several genetic variants in the 9p21 region have been identified that are associated with the risk of multiple cancers. Here, we explored the association of two genetic variants in the 9p21 region, CDKN2A/B, rs10811661, and rs1333049 for the first time in 273 subjects with, or without ESCC. We observed that the patients with ESCC had a higher frequency of a TT genotype for rs10811661 than individuals in the control group, and this polymorphism was also associated with tumor size. Moreover, a CC genotype for the rs1333049 polymorphism was associated with a reduced overall survival (OS) of patients with ESCC. In particular, patients with a CC (rs1333049) genotype had a significantly shorter OS (CC genotype: 34.5 ± 8.9 months vs. CG+GG: 47.7 ± 5.9 months; \np value = 0.03). We have also shown the association of a novel genetic variant in CDKN2B gene with clinical outcome of patients with ESCC. Further investigations are warranted in a larger population to explore the value of emerging markers as a risk stratification marker in ESCC." - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27310   open full text
• The genetic factors contributing to hypospadias and their clinical utility in its diagnosis.
  Marjan Joodi, Forouzan Amerizadeh, Seyed Mahdi Hassanian, Marjan Erfani, Majid Ghayour‐Mobarhan, Gordon A. Ferns, Majid Khazaei, Amir Avan.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Hypospadias is among the most common congenital malformations in male neonates. It results from abnormal penile and urethral development, but is a multifactorial disorder that is highly heterogeneous, with several genetic and environmental determinants. Monogenic and chromosomal abnormalities are present in approximately 30% of cases, although the genetic factors contributing to hypospadias remain unknown in 70% of cases. While defects in androgen synthesis can lead to this malformation, mutational analyses have shown several genes, such as sonic hedgehog, fibroblast growth factors, bone morphogenetic proteins, homeobox genes, and the Wnt family, are involved in the normal development of male external genitalia. Mutations in the genes of penile development (e.g., HOX, FGF, Shh) and testicular determination (e.g., WT1, SRY), luteinizing hormone receptor, and androgen receptor have also been proposed to be implicated in hypospadias. Here we review the recent advances in this field and discuss the potential genes that could determine the risk of hypospadias. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27350   open full text
• Curcumin: A naturally occurring autophagy modulator.
  Abolfazl Shakeri, Arrigo F. G. Cicero, Yunes Panahi, Mohammad Mohajeri, Amirhossein Sahebkar.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Autophagy is a self‐degradative process that plays a pivotal role in several medical conditions associated with infection, cancer, neurodegeneration, aging, and metabolic disorders. Its interplay with cancer development and treatment resistance is complicated and paramount for drug design since an autophagic response can lead to tumor suppression by enhancing cellular integrity and tumorigenesis by improving tumor cell survival. In addition, autophagy denotes the cellular ability of adapting to stress though it may end up in apoptosis activation when cells are exposed to a very powerful stress. Induction of autophagy is a therapeutic option in cancer and many anticancer drugs have been developed to this aim. Curcumin as a hydrophobic polyphenol compound extracted from the known spice turmeric has different pharmacological effects in both in vitro and in vivo models. Many reports exist reporting that curcumin is capable of triggering autophagy in several cancer cells. In this review, we will focus on how curcumin can target autophagy in different cellular settings that may extend our understanding of new pharmacological agents to overcome relevant diseases. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27404   open full text
• Iturin A‐like lipopeptides from Bacillus subtilis trigger apoptosis, paraptosis, and autophagy in Caco‐2 cells.
  Haobin Zhao, Xiaoguang Xu, Shuzhen Lei, Dongyan Shao, Chunmei Jiang, Junling Shi, Yawen Zhang, Li Liu, Shuzhen Lei, Hui Sun, Qingsheng Huang.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract This study revealed that iturin A‐like lipopeptides produced by Bacillus subtillis induced both paraptosis and apoptosis in heterogeneous human epithelial colorectal adenocarcinoma (Caco‐2) cells. Autophagy was simultaneously induced in Caco‐2 cells treated with iturin A‐like lipopeptides at the early stage and inhibited at the later stage. A western blot analysis showed that the lipopeptides induced apoptosis in Caco‐2 cells via a mitochondrial‐dependent pathway, as indicated by upregulated expression of the apoptotic genes bax and bad and downregulated expression of the antiapoptotic gene bcl‐2. The induction of paraptosis in Caco‐2 cells was indicated by the occurrence of many cytoplasmic vacuoles accompanied by endoplasmic reticulum (ER) dilatation and mitochondrial swelling and dysfunction. ER stress also occurred with significant increases in reactive oxygen species and Ca2+ levels in cells. Autophagy was detected by a transmission electron microscopy analysis and by upregulated expression of LC3‐II and downregulated expression of LC3‐I. The inhibition of autophagy at the later stage was shown by upregulated expression of p62. This study revealed the capability of iturin A‐like B. subtilis lipopeptides to simultaneously execute antitumor potential via multiple pathways. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27377   open full text
• Laminar shear stress‐provoked cytoskeletal changes are mediated by epigenetic reprogramming of TIMP1 in human primary smooth muscle cells.
  Rodrigo A. da Silva, Célio Jr da C. Fernandes, Geórgia da S. Feltran, Anderson M. Gomes, Amanda Fantini Andrade, Denise C. Andia, Maikel P. Peppelenbosch, Willian F. Zambuzzi.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2 Abstract Whereas endothelial responses to shear stress are well‐characterized, the cell physiological effects of shear stress in smooth muscle cells (SMCs) remain largely obscure. As SMCs are directly challenged by shear stress after endothelial denuding injury following procedures such as angioplasty or endarterectomy, characterization of these responses represents an important scientific question. Hence we decided to contrast cytoskeletal reorganization, epigenetic reprogramming, signaling transduction, and changes in miRNA (miRs) profiles in primary human aortic smooth muscle cells (AoSMCs) between unstressed cells and cells exposed to shear stress. We observed that shear stress‐provoked reorganization of the actin cytoskeleton in an apparently Cofilin‐dependent fashion and which related to altered integrin signaling, apparently caused by remodeling of the extracellular matrix. The latter appeared a downstream effect of increased expression of matrix metalloproteinases and downregulation of tissue metalloproteinase inhibitor 1 (TIMP1) protein levels. In turn, these effects related to shear stress‐provoked changes in expression and nuclear localization of the epigenetic regulators demethylases TET1, TET2, DNMT1, DNMT3A and DNMT3B, HDAC6, and SIRT1. Accordingly, TIMP1 promotor CpG hypomethylation was a prominent effect, and resulted in a significant increase in TIMP1 transcription, which may also have related increased expression of miRs involved in modulating TIMP1 translation. Thus epigenetic‐reprogramming of TIMP1 emerges as critical element in smooth muscle responses to mechanical signals and as epigenetic machinery is amendable to pharmacological manipulation, this pathway may have important clinical consequences. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27374   open full text
• Exercise rescues the immune response fine‐tuned impaired by peroxisome proliferator‐activated receptors γ deletion in macrophages.
  Loreana Sanches Silveira, Helena Angélica Pereira Batatinha, Angela Castoldi, Niels Olsen Saraiva Câmara, Willian T. Festuccia, Camila Oliveira Souza, José Cesar Rosa Neto, Fábio Santos Lira.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2 Abstract Background Exercise is a powerful tool for prevention and treatment of many conditions related to the cardiovascular system and also chronic low‐grade inflammation. Peroxisome proliferator‐activated receptors γ (PPARγ) exerts an import role on the regulation of metabolic profile and subsequent inflammatory response, especially in macrophages. Purpose To investigate the effects of 8‐week moderate‐exercise training on metabolic and inflammatory parameters in mice with PPARγ deficiency in myeloid cells. Methods Twelve‐week old mice bearing PPARγ deletion exclusively in myeloid cells (PPARγlox/lox Lys Cre −/+, knockout [KO]) and littermate controls (PPARγlox/lox Lys Cre −/−, wild type [WT]) were submitted to 8‐week exercise training (treadmill running at moderate intensity, 5 days/week). Animals were evaluated for food intake, glucose homeostasis, serum metabolites, adipose tissue and peritoneal macrophage inflammation, and basal and stimulated cytokine secretion. Results Exercise protocol did not improve glucose metabolism or adiponectin concentrations in serum of KO mice. Moreover, the absence of PPARγ in macrophages exacerbated the proinflammatory profile in sedentary mice. Peritoneal cultured cells had higher tumor necrosis factor‐α (TNF‐α) secretion in nonstimulated and lipopolysaccharide (LPS)‐stimulated conditions and higher Toll‐4 receptor (TLR4) gene expression under LPS stimulus. Trained mice showed reduced TNF‐α content in adipose tissue independently of the genotype. M2 polarization ability was impaired in KO peritoneal macrophages after exercise training, while adipose tissue‐associated macrophages did not present any effect by PPARγ ablation. Conclusion Overall, PPARγ seems necessary to maintain macrophages appropriate response to inflammatory stimulus and macrophage polarization, affecting also whole body lipid metabolism and adiponectin profile. Exercise training showed as an efficient mechanism to restore the immune response impaired by PPARγ deletion in macrophages. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27333   open full text
• Stromal vascular fraction cells plus sustained release VEGF/Ang‐1‐PLGA microspheres improve fat graft survival in mice.
  Yucang He, Xiaofang Yu, Zhuojie Chen, Liqun Li.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Autologous fat transplantation is increasingly applied in plastic and reconstructive surgery. Stromal vascular fraction cells (SVFs) combined with angiogenic factors, such as VEGF (vascular endothelial growth factor A) and Ang‐1 (angiogenin‐1), can improve angiogenesis, which is a critical factor for graft survival. However, direct transplant with such a mixture is insufficient owing to the short half‐life of angiogenic factors. In this study, we evaluated whether a double sustained release system of VEGF/ANG‐1‐PLGA (poly (lactic‐co‐glycolic acid)) microspheres plus SVFs can improve angiogenesis and graft survival after autologous fat transplantation. VEGF/ANG‐1‐PLGA‐sustained release microspheres were fabricated by a modified double emulsion–solvent evaporation technique. Human aspirated fat was mixed with SVF suspension plus VEGF/ANG‐1 sustained release microspheres (Group C), SVF suspension (Group B) alone, or Dulbecco’s modified Eagle’s medium as the control (Group A). Eighteen immunocompromised nude mice were injected with these three mixtures subcutaneously at random positions. After 8 weeks, the mean volume of grafts was greater in the SVFs plus VEGF/ANG‐1‐PLGA group than in the control and SVFs groups (1.08 ± 0.069 ml vs. 0.62 ± 0.036 ml, and 0.83 ± 0.059 ml, respectively). Histological assessments showed that lower fibrosis, but greater microvascular density in the SVFs plus VEGF/ANG‐1‐PLGA group than in the other groups, though the SVFs group also had an appropriate capillary density and reduced fibrosis. Our findings indicate that SVFs plus VEGF/ANG‐1‐PLGA‐sustained release microspheres can improve angiogenesis and graft survival after autologous fat transplantation. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27368   open full text
• Ephrin A1 promotes proliferation of bovine endometrial cells with abundant expression of proliferating cell nuclear antigen and cyclin D1 changing the cell population at each stage of the cell cycle.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Ephrin A1 has a role in a variety of biological events, including cell proliferation, differentiation, migration, and angiogenesis. Ephrin A1 expression is abundant in trophoblasts and endometrial cells during the implantation period; however, its intracellular activities have not yet been reported in bovine endometrial (BEND) epithelial cells. The aim of this study was to identify the functional role of ephrin A1 in BEND cells, which have served as a good model system for investigating the regulation of signal transduction following treatment with interferon‐τ (IFNT) in vitro. Supplementation of ephrin A1 to BEND cells increased cell proliferation and increased levels of proliferating cell nuclear antigen and cyclin D1 protein in BEND cell nuclei. To investigate intracellular mechanisms regulated by ephrin A1, we performed Western blot analysis focused on mitogen‐activated protein kinase (MAPK) and phosphoinositide 3‐kinase (PI3K) signaling, which are significantly involved in the successful maintenance of pregnancy. Ephrin A1 dose‐dependently increased phosphorylation of extracellular signal‐regulated kinases (ERK)1/2, c‐Jun N‐terminal kinases (JNK), P38, protein kinase B (AKT), P70S6K, S6, and cyclin D1, and the activated proteins were suppressed by pharmacological inhibitors including wortmannin (a PI3K inhibitor), U0126 (an ERK1/2 inhibitor), and SP600125 (a JNK inhibitor). Among ephrin A1 receptors, abundant expression of EPHA2 and EPHA4 messenger RNA was detected in BEND cells by reverse transcription polymerase chain reaction analysis. Furthermore, tunicamycin‐induced endoplasmic reticulum (ER) stress was inactivated by ephrin A1 treatment of BEND cells. Our findings suggest that ephrin A1 promotes the development of BEND cells and likely enhances uterine capacity and maintenance of pregnancy by activating MAPK and PI3K signaling cascades and by restoring ER stress. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27275   open full text
• Molecular mechanisms involved in the protective effect of pituitary adenylate cyclase‐activating polypeptide in an in vitro model of amyotrophic lateral sclerosis.
  Grazia Maugeri, Agata G. D’Amico, Daniela M. Rasà, Concetta Federico, Salvatore Saccone, Giovanna Morello, Valentina Cognata, Sebastiano Cavallaro, Velia D’Agata.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the loss of upper and lower motor neurons. Based on transcriptional profiles of motor cortex samples, in a previous work, we were able to classify two subgroups of sporadic ALS (SALS) patients, named SALS1 and SALS2. A further meta‐analysis study has revealed sixteen drug targets commonly deregulated in SALS2 and superoxide dismutase 1 (SOD1) G93A mice. The identified candidate drug targets included pituitary adenylate cyclase‐activating polypeptide (PACAP), epidermal growth factor receptor (EGFR) and matrix metallopeptidase‐2 (MMP‐2). By using a motor neuron‐like hybrid cell line (NSC‐34) expressing human SOD1 G93A as an in vitro model of ALS, here we investigated the functional correlation among these three genes. Our results have shown that PACAP increases cell viability following serum deprivation. This effect is induced through EGFR transactivation mediated by protein kinase A stimulation. Furthermore, EGFR phosphorylation activates mitogen‐activated protein kinases/extracellular signal‐regulated kinases 1 and 2 survival signaling pathway and increases MMP‐2 expression, significantly reduced by serum starvation. These results suggest that a deeper characterization of mechanisms involved in PACAP/EGFR/MMP‐2 axis activation in G93A SOD1 mutated neurons may allow identifying new targets for ALS therapy. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27328   open full text
• Teeth‐derived stem cells: A source for cell therapy.
  Ghazaleh Baniebrahimi, Razieh Khanmohammadi, Fatemeh Mir.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2 Abstract Cell therapy is one of the important therapeutic approaches in the treatment of many diseases such as cancer, degenerative diseases, and cardiovascular diseases. Among various cell types, which could be used as cell therapies, stem cell therapy has emerged as powerful tools in the treatment of several diseases. Multipotent stem cells are one of the main classes of stem cells that could originate from different parts of the body such as bone marrow, adipose, placenta, and tooth. Among several types of multipotent stem cells, tooth‐derived stem cells (TDSCs) are associated with special properties such as accessible, easy isolation, and low invasive, which have introduced them as a good source for using in the treatment of several diseases such as neural injuries, liver fibrosis, and Cohrn’s disease. Here, we provided an overview of TDSCs particular stem cells from human exfoliated deciduous teeth and clinical application of them. Moreover, we highlighted molecular mechanisms involved in the regulation of dental stem cells fate. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27270   open full text
• Positive feedback loop of lncRNA LINC01296/miR‐598/Twist1 promotes non‐small cell lung cancer tumorigenesis.
  Lijuan Xu, Bin Wei, Hongxia Hui, Yuan Sun, Yangqing Liu, Xiaojuan Yu, Jian Dai.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Emerging evidence has illustrated the vital roles of long noncoding RNAs (lncRNAs) in human cancers. However, the role of lncRNAs in non‐small cell lung cancer (NSCLC) is still elusive and poorly understood. In the current study, our team conducted extensive experiments to identify the role of long intergenic nonprotein coding (LINC01296) on NSCLC tumorigenesis. The results illustrated that the elevated LINC01296 expression in NSCLC tissue specimens and cell lines were closely correlated with the poor prognosis of patients with NSCLC. Functional studies revealed that LINC01296 knockdown silenced by small interfering RNAs inhibited proliferation, accelerated apoptosis in vitro, and impaired tumor growth in vivo. Mechanical studies showed that INC01296 harbored miR‐598, acting as a microRNA “sponge.” Besides, miR‐598 targeted the 3′‐UTR of Twist1. Interestingly, transcription factor Twist1 could bind with the promoter of INC01296 and activate its transcriptional level. In summary, we conclude that INC01296/miR‐598/Twist1 constitutes a positive feedback loop to promote the tumorigenesis of NSCLC, providing a novel insight and a valuable therapeutic strategy. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27235   open full text
• Glucocorticoids reduce chemotherapeutic effectiveness on OSCC cells via glucose‐dependent mechanisms.
  Antonio Celentano, Michael McCullough, Nicola Cirillo.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Synthetic corticosteroids are routinely administered during the treatment of several diseases, including malignancies. However, recent evidence suggests that corticosteroids may have tumor‐promoting effects, particularly in epithelial neoplasms. Our aim was to assess the role of the recently characterized cancer‐associated glucocorticoid (GC) system in the resistance to chemotherapy of oral malignant keratinocytes. Human malignant oral keratinocyte cell lines H314/H357/H400/BICR16/BICR56 were tested with: two chemotherapeutic agents, doxorubicin (DOXO) and 5‐fluorouracil (5‐FU), as well as hydrocortisone (HC), adrenocorticotropic hormone (ACTH), 5‐pregnen‐3‐beta‐ol‐20‐one‐16‐alfa‐carbonitrile (PCN), and two glucose uptake inhibitors, Fasentin and WZB. Both DOXO and 5‐FU induced apoptosis in a dose‐dependent and time‐dependent manner. HC administration (100 nM) reduced the effectiveness of both chemotherapeutic agents to a variable extent in all 5 oral squamous cell carcinoma cell lines. ACTH also reduced the effectiveness of DOXO on 2 cell lines tested (H357 and BICR56). The glucose uptake inhibitors Fasentin and WZB were able to partially block the increased resistance to the cytotoxic drugs induced by HC. In summary, we have demonstrated, for the first time, the importance of cortisol on oral cancer cells ability to proliferate and combat the effectiveness of chemotherapeutic agents. This effect appears to be glucose dependent. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27227   open full text
• In vivo microscopic and optical coherence tomography classification of neurotrophic keratopathy.
  Leonardo Mastropasqua, Mario Nubile, Manuela Lanzini, Roberta Calienno, Harminder S. Dua.
  Journal of Cellular Physiology. September 21, 2018
  --- - |2- Abstract Neurotrophic keratopathy (NK) is a rare degenerative corneal disorder characterized by instability of epithelial integrity with consequent epithelial defects that can worsen up to persistent epithelial defects with stromal melting and ulceration. The pathogenesis of NK springs from a variable degree of damage to the trigeminal nerve plexus, leading to a reduction or total loss of corneal sensitivity. Mackie classification (1995) distinguishes three stages of NK, based on the severity of clinical presentation. The technological innovations in corneal diagnostic imaging allow clinicians to accurately study the morphometry and morphology of corneal structure with microscopic resolution. In this study, 45 patients affected by NK at different stages underwent in vivo confocal microscopy (IVCM) and anterior segment optical coherence tomography (AS‐OCT) with particular attention to analyze subbasal nerve plexus fibers and the stromal structure. At the light of IVCM and AS‐OCT observations, we propose a different staging of NK with respect to the Mackie's classification that takes into account the severity of subbasal nerve fibers damage and the extension in depth of stromal ulceration; this classification better defines, at the time of diagnosis, the cellular and structural alterations in the affected corneas, with possible prognostic and therapeutic values in the management of NK. - Journal of Cellular Physiology, EarlyView.
  

  September 21, 2018   doi: 10.1002/jcp.27345   open full text
• Pyruvate dehydrogenase kinase 1 contributes to cisplatin resistance of ovarian cancer through EGFR activation.
  Meng Zhang, Qing Cong, Xiao‐Yan Zhang, Ming‐Xing Zhang, Ying‐Ying Lu, Cong‐Jian Xu.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract Patients with ovarian cancer frequently develop acquired drug resistance after the long‐term chemotherapy, leading to disease progression. Enhanced epithelial–mesenchymal transition (EMT) has been implicated in chemoresistance of ovarian cancer cells; however, the molecular mechanisms involved are largely undefined. Pyruvate dehydrogenase kinase 1 (PDK1), a key regulatory enzyme in glucose metabolism, has been recognized as a gatekeeper of the Warburg effect, a hallmark of cancer. In this study, the function of PDK1 in cisplatin resistance of ovarian cancer in terms of growth and EMT was investigated. PDK1 was upregulated in cisplatin‐resistant ovarian cancer cells. PDK1 knockdown in resistant cells led to increased sensitivity to cisplatin‐induced cell death and apoptosis. PDK1 downregulation also reversed the EMT and cell motility in cisplatin‐resistant cells. In a mouse xenograft model, tumors derived from PDK1‐silenced ovarian cancer cells exhibited decreased tumor growth and EMT compared with control after the cisplatin treatment. Mechanistically, PDK1 overexpression led to increased phosphorylation of EGFR, and blocking EGFR kinase activity by erlotinib reversed cisplatin resistance induced by PDK1 overexpression. Furthermore, in patients with ovarian cancer, higher PDK1 and p‐EGFR levels were associated with chemoresistance. These results supported that PDK1 contributes to chemoresistance of ovarian cancer by activating EGFR. Therefore, PDK1 may serve as a promising target to combat chemoresistance of ovarian cancer. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27369   open full text
• Association of the genetic polymorphisms in immunoinflammatory microRNAs with risk of ischemic stroke and subtypes in an Iranian population.
  Hassan Darabi, Arash Salmaninejad, Mohamad Ehsan Jaripour, Mahmoud reza Azarpazhooh, Majid Mojarrad, Ariane Sadr‐Nabavi.
  Journal of Cellular Physiology. September 19, 2018
  --- - "\nAbstract\nStroke is one of the most common type of cerebrovascular disease threatening human health and life with high mortality, disability, and morbidity. Ischemic stroke (IS) is determined to be a complex disease containing a group of heterogeneous disorders with various environmental and genetic risk factors. This study evaluated the polymorphisms of microRNAs involved in inflammatory routes leading to stroke in an Iranian population. This study evaluated the associations of hsa‐mir‐608 C/G rs4919510, hsa‐mir‐499 A/G rs3746444, and hsa‐mir‐145 C/T rs190323149 polymorphisms in precursor miRNAs with the risk of IS. These microRNA polymorphisms were analyzed in 470 patients with IS and 489 control subjects. The TOAST criteria was applied for IS subtypes classification. The frequency of the allele G of hsa‐mir‐499/rs3746444 A/G revealed significant association with IS in comparison with controls (\np < 0.0001, OR = 1.838, 95% CI = 1.406–2.401). Increased IS risks were associated with hsa‐mir‐499/ rs3746444 A/G genotypes in diverse genetic model (homozygote comparison: \np = 0.004, OR = 2.136, 95% CI = 1.269–3.597; heterozygote comparison: \np = 0.029, OR = 1.373, 95% CI = 1.033–1.825). Statistical analysis in IS subtypes showed that cardio‐embolic patients compared with other subtypes (large artery atherosclerosis and lacunar) had higher frequency of G allele (LAA vs. CEI, \np = 0.017; LAC vs. CEI, \np = 0.009), AG genotype (LAA vs. CEI, \np = 0.016; LAC vs. CEI, \np = 0.013). Nevertheless, this study did not find any association between the alleles and genotypes of mir‐608 C/G rs4919510 SNP and IS, respectively (\np > 0.05). The current investigation provided verification that hsa‐mir‐499 rs3746444 A/G polymorphism may be associated with a significantly increased risk of IS in an Iranian population.\n" - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27159   open full text
• Melatonin and pancreatic cancer: Current knowledge and future perspectives.
  Omid Reza Tamtaji, Naghmeh Mirhosseini, Russel J. Reiter, Morteza Behnamfar, Zatollah Asemi.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2 Abstract Pancreatic cancer has a high mortality rate due to the absence of early symptoms and subsequent late diagnosis; additionally, pancreatic cancer has a high resistance to radio‐ and chemotherapy. Multiple inflammatory pathways are involved in the pathophysiology of pancreatic cancer. Melatonin an indoleamine produced in the pineal gland mediated and receptor‐independent action is the pancreas and other where has both receptors. Melatonin is a potent antioxidant and tissue protector against inflammation and oxidative stress. In vivo and in vitro studies have shown that melatonin supplementation is an appropriate therapeutic approach for pancreatic cancer. Melatonin may be an effective apoptosis inducer in cancer cells through regulation of a large number of molecular pathways including oxidative stress, heat shock proteins, and vascular endothelial growth factor. Limited clinical studies, however, have evaluated the role of melatonin in pancreatic cancer. This review summarizes what is known regarding the effects of melatonin on pancreatic cancer and the mechanisms involved. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27372   open full text
• New advances of lncRNAs in liver fibrosis, with specific focus on lncRNA–miRNA interactions.
  Er‐Bao Bian, Zhi‐Gang Xiong, Jun Li.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract Noncoding RNAs (ncRNAs) were initially thought to be transcriptional byproducts. However, recent advances of ncRNAs research have increased our understanding of the importance of ncRNA in gene regulation and disease pathogenesis. Consistent with these developments, liver fibrosis research is also experiencing rapid growth in the investigation of links between ncRNAs and the pathology of this disease. The initial focus was on studying the function and regulation mechanisms of microRNAs (miRNAs). However, recently, elucidation of the mechanisms of long noncoding RNAs (lncRNAs) and lncRNA‐mediated liver fibrosis has just commenced. In this review, we emphasize on abnormal expression of lncRNAs in liver fibrosis. Furthermore, we also discuss that the interaction of lncRNAs with miRNAs is involved in the regulation of the expression of protein‐coding genes in liver fibrosis. Recent advances in understanding dysregulated lncRNAs expression and the lncRNAs–miRNAs interaction in liver fibrosis will help for developing new therapeutic targets and biomarkers of liver fibrosis. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27069   open full text
• MicroRNA‐363 inhibits angiogenesis, proliferation, invasion, and migration of renal cell carcinoma via inactivation of the Janus tyrosine kinases 2–signal transducers and activators of transcription 3 axis by suppressing growth hormone receptor gene.
  Jie Zhu, Da‐Qing Zhu, Yu Zhang, Qi‐Ming Liu, Peng‐Chao Wang, Hong‐Zhao Li, Xin Ma, Xu Zhang.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract Renal cell carcinoma (RCC) is the most common malignancy involving the kidneys and a major cause of cancer mortality. The involvement of microRNA (miRNA) expression in the tumorigenesis and progression of RCC has been previously highlighted. Therefore, we conducted this study to investigate whether microRNA‐363 (miR‐363) affects the development of RCC via the Janus tyrosine kinases (JAK2)–signal transducers and activators of transcription (STAT) axis by targeting the growth hormone receptor (GHR), by observing the changes that occurred in the RCC and the normal adjacent tissues of patients with RCC. RCC cells were transfected with a series of miR‐363 mimic, miR‐363 inhibitor, or small interfering RNA against GHR to determine the influence of miR‐363 on the expression of GHR and JAK2–STAT3 axis‐related genes with the use of reverse transcription quantitative polymerase chain reaction and Western blot analysis. The angiogenesis, viability, invasion, and migration of cells were evaluated by means of in vitro angiogenesis, 3‐(4,5)‐dimethylthiahiazo (‐z‐y1)‐3,5‐di‐phenytetrazoliumromide (MTT), wound‐healing, and Transwell assays. The results revealed reduced miR‐363 expression and elevated GHR expression in RCC. It was also found that miR‐363 altered the activation of the JAK2–STAT3 axis through the inhibition of GHR. Cells treated with the miR‐363 inhibitor presented with increased capillary vessels, cell viability, invasion, and migration, whereas it was on the contrary in the RCC cells with overexpressed miR‐363. These results implicated that the overexpression of miR‐363 could specifically bind to GHR to downregulate the expression of GHR, which, in turn, inactivates the JAK2–STAT3 axis, thereby influencing the angiogenesis, cell invasion, and migration abilities in RCC. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27020   open full text
• MiR‐499 regulates myoblast proliferation and differentiation by targeting transforming growth factor β receptor 1.
  Jiyao Wu, Binglin Yue, Xianyong Lan, Yanhuan Wang, Xingtang Fang, Yun Ma, Yueyu Bai, Xingshan Qi, Chunlei Zhang, Hong Chen.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract MicroRNAs (miRNAs or miRs) are small noncoding RNAs that play critical roles in muscle cell proliferation and differentiation via post‐transcriptional regulation of gene expression. Here, based on our previous high‐throughput sequencing results, we evaluated miRNA‐499 (miR‐499) functions during myoblast proliferation and differentiation. In addition, we analyzed miR‐499 expression profiles and characterized the associated functional roles. MiR‐499 is known to be a skeletal muscle fiber‐type‐associated miRNA. However, its roles in skeletal myoblast proliferation and differentiation are poorly understood. MiR‐499 overexpression promoted C2C12 cell proliferation and significantly attenuated C2C12 cell myogenic differentiation. Furthermore, miR‐499 inhibition enhanced C2C12 cell proliferation and suppressed C2C12 cell differentiation. Using dual‐luciferase reporter assays and western blot analysis, we confirmed that miR‐499 targeted transforming growth factor β receptor 1 (TGFβR1), a known regulator of skeletal myoblast development. Additionally, our RNA interference analysis, in which TGFβR1 was downregulated, showed that TGFβR1 significantly promoted the differentiation of C2C12 cells and inhibited their proliferation. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.26903   open full text
• Chloride channel‐3 mediates multidrug resistance of cancer by upregulating P‐glycoprotein expression.
  Qi Chen, Xueqiang Liu, Zhesi Luo, Shisi Wang, Jialin Lin, Zheng Xie, Mengge Li, Chunmei Li, Hua Cao, Qingsong Huang, Jianwen Mao, Bin Xu.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract Chloride channel‐3 (ClC‐3), a member of the ClC family of voltage‐gated Cl− channels, is involved in the resistance of tumor cells to chemotherapeutic drugs. Here, we report a new mechanism for ClC‐3 in mediating multidrug resistance (MDR). ClC‐3 was highly expressed in the P‐glycoprotein (P‐gp)‐dependent human lung adenocarcinoma cell line (A549)/paclitaxel (PTX) and the human breast carcinoma cell line (MCF‐7)/doxorubicin (DOX) resistant cells. Changes in the ClC‐3 expression resulted in the development of drug resistance in formerly drug‐sensitive A549 or MCF‐7 cells, and drug sensitivity in formerly drug‐resistant A549/Taxol and MCF‐7/DOX cells. Double transgenic MMTV‐PyMT/CLCN3 mice with spontaneous mammary cancer and ClC‐3 overexpression demonstrated drug resistance to PTX and DOX. ClC‐3 expression upregulated the expression of MDR1 messenger RNA and P‐gp by activating the nuclear factor‐κB (NF‐κB)‐signaling pathway. These data suggest that ClC‐3 expression in cancer cells induces MDR by upregulating NF‐κB‐signaling‐dependent P‐gp expression involving another new mechanism for ClC‐3 in the development of drug resistance of cancers. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27402   open full text
• Slug mediates myofibroblastic differentiation to promote fibrogenesis in buccal mucosa.
  Chih‐Yuan Fang, Shih‐Min Hsia, Pei‐Ling Hsieh, Yi‐Wen Liao, Chih‐Yu Peng, Ching‐Zong Wu, Kuan‐Chou Lin, Lo‐Lin Tsai, Cheng‐Chia Yu.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract Epithelial–mesenchymal transition (EMT) has been implicated in fibrogenesis and carcinogenesis; however, the exact role of EMT‐inducer Slug in the progression of precancerous oral submucous fibrosis (OSF) has not been investigated. In the current study, we showed that the expression of Slug was upregulated in OSF tissues and associated with various myofibroblast markers. After silence of Slug in fibrotic buccal mucosal fibroblasts (fBMFs), the elevated myofibroblast activities and fibrosis markers were all downregulated. Our data revealed that arecoline, an areca nut alkaloid, increased the expression of Slug in normal BMFs, and inhibition of Slug successfully prevented the arecoline‐induced myofibroblast activation. Additionally, overexpression of Slug in BMFs stimulated the activities of myofibroblasts, indicating that upregulation of Slug by arecoline contributes to the myofibroblast transdifferentiation. Most importantly, Slug was able to bind to the E‐box of type I collagen, leading to increased expression of type I collagen. Altogether, this study demonstrated the abnormal elevation of Slug in OSF and its significance in arecoline‐induced fibrogenesis. Moreover, downregulation of Slug could be a potential target for OSF remedy via suppression of myofibroblast activities and type I collagen. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27418   open full text
• Efficient penetration of Scp01‐b and its DNA transfer abilities into cells.
  Ming Zhang, Xueli Zhao, Jingping Geng, Huiting Liu, Fanhui Zeng, Yanyan Qin, Jason Li, Changbai Liu, Hu Wang.
  Journal of Cellular Physiology. September 19, 2018
  --- - |2- Abstract The in vivo application potential of viral‐based gene delivery approaches is hindered by a risk of insertional oncogenesis. Of the many delivery methods, cell‐penetrating peptides (CPP)‐based delivery has good biocompatibility and biodegradability. However, low efficiency is still the disadvantage of CPPs‐based nucleic acid transfection, and delivery efficiency may vary from different CPPs. Here, we describe Scp01‐b, as a new CPP, which can enter cultured cell lines and primary cultured cells examined by fluorescence microscopy and quantitative assay, the internalization process is a concentration, temperature, and incubation time‐dependent manner. Scp01‐b does not insert into the membrane directly and its uptake is mediated through endocytosis pathway. Moreover, Scp01‐b could mediate the uptake of plasmid DNA into the Caski and HSC‐T6 cells, and we noted that Scp01‐b‐mediated transfection efficiency was nearly the same with traditional liposome (TurboFectin)‐mediated transfection. These findings suggest that Scp01‐b can act as a useful tool for non‐viral‐based delivery in further application such as reprogramming and gene editing. - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27392   open full text
• LncRNA CRNDE promotes hepatocellular carcinoma cell proliferation, invasion, and migration through regulating miR‐203/ BCAT1 axis.
  Degang Ji, Chengwei Jiang, Lirong Zhang, Na Liang, Tiechao Jiang, Bin Yang, Haiying Liang.
  Journal of Cellular Physiology. September 19, 2018
  --- - "\nAbstract\n\nObjective\nTo investigate the impact of long noncodingRNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) on hepatocellular cancer (HCC) cell propagation, invasion, and migration by mediating miR‐203/\nBCAT1 axis.\n\n\nMethods\nMicroarray analysis was based on 25 pairs of HCC cancerous tissues and adjacent tissues. The expression levels of CRNDE, miR‐203, and BCAT1 in HCC tissues were analyzed by quantitative real‐time polymerase chain reaction (qRT‐PCR). The liver cell line L‐02 and HCC cell lines HepG2 and Huh‐7 were utilized to assess the regulatory effects of CRNDE and miR‐203 on HCC progression in vitro. Western blot was used to qualify BCAT1 protein expression level. Cell proliferation and apoptosis were evaluated using CCK‐8 and flow cytometry analysis, whereas cell invasion and migration assay were performed by the Transwell assay. The relationship among CRNDE, miR‐203, and \nBCAT1 was validated by dual luciferase assay. Tumor Xenograft study was established to verify the pathological effect of CRNDE on HCC development in vivo.\n\n\nResults\nThe expression levels of the CRNDE and BCAT1 were upregulated in HCC tissues and cells, whereas miR‐203 was downregulated in HCC. Knockdown of CRNDE or miR‐203 overexpression would inhibit HCC cell propagation and metastasis, and induced cell apoptosis. Moreover, miR‐203 was negatively correlated with CRNDE, the same as miR‐203 with \nBCAT1. Dual luciferase assay showed that miR‐203 was an inhibitory target of CRNDE, and \nBCAT1 was directly targeted by miR‐203 as well.\n\n\nConclusion\nLncRNA CRNDE could enhance HCC tumorgenesis by sponging miR‐203 and mediating BCAT1. LncRNA CRNDE might facilitate HCC cell propagation, invasiveness, and migration through regulating miR‐203/\nBCAT1 axis.\n" - Journal of Cellular Physiology, EarlyView.
  

  September 19, 2018   doi: 10.1002/jcp.27396   open full text
• The metabolic and molecular mechanisms of hyperammonaemia‐ and hyperethanolaemia‐induced protein catabolism in skeletal muscle cells.
  Hannah Crossland, Kenneth Smith, Philip J. Atherton, Daniel J. Wilkinson.
  Journal of Cellular Physiology. August 24, 2018
  --- - |2- Hyperammonaemia and hyperethanolaemia are thought to be driving factors behind skeletal muscle myopathy in liver disease, that is, cirrhosis. Despite this, the singular and combined impacts of ethanol‐ and ammonia‐induced protein catabolism are poorly defined. As such, we aimed to dissect out the effects of ammonia and ethanol on muscle catabolism. Murine C2C12 myotubes were treated with ammonium acetate (10 mM) and ethanol (100 mM) either alone or in combination for 4 hr and/or 24 hr. Myotube diameter, muscle protein synthesis and anabolic and catabolic signalling pathways were assessed. In separate experiments, cells were cotreated with selected inhibitors of protein breakdown to assess the importance of proteolytic pathways in protein loss with ammonia and ethanol. Ammonia and ethanol in combination resulted in a reduction in myotube width and total protein content, which was greater than the reduction observed with ammonia alone. Both ammonia and ethanol caused reductions in protein synthesis, as assessed by puromycin incorporation. There was also evidence of impairments in regulation of protein translation, and increased protein expression of markers of muscle protein breakdown. Myotube protein loss with ammonia plus ethanol was not affected by autophagy inhibition, but was completely prevented by proteasome inhibition. Thus, combined ammonia and ethanol incubation of C2C12 myotubes exacerbated myotube atrophy and dysregulation of anabolic and catabolic signalling pathways associated with either component individually. Ubiquitin proteasome‐mediated protein breakdown appears to play an important role in myotube protein loss with ethanol and ammonia. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9663-9673, December 2018.
  

  August 24, 2018   doi: 10.1002/jcp.26881   open full text
• MicroRNA‐16 functions as a tumor‐suppressor gene in oral squamous cell carcinoma by targeting AKT3 and BCL2L2.
  Xi Wang, Guang‐hui Li.
  Journal of Cellular Physiology. August 22, 2018
  --- - |2 Aberrant expressions of microRNAs have been reported to be strongly associated with the progression and prognosis of various tumors, including oral squamous cell carcinoma (OSCC). Recent studies on miRNA expression profiling have suggested that microRNA‐16 (miR‐16) may be dysregulated in OSCC. However, the tumorigenic roles and mechanisms of miR‐16 in OSCC are still largely unknown. In this study, we demonstrated that miR‐16 was specifically downregulated in both OSCC patients and cancer cell lines. In addition, functional roles of miR‐16 in vitro suggested that the miR‐16 mimic inhibited cell proliferation and induced apoptosis, whereas miR‐16 inhibitor displayed the opposite effects. Luciferase reporter assay and correlation analysis showed that AKT3 and BCL2L2 were directly targeted by miR‐16 and were inversely expressed with miR‐16 in OSCC. Moreover, restoration of AKT3 and BCL2L2 expression could partially reverse the cell proliferation inhibition and apoptosis induction caused by miR‐16. In xenograft nude mice, miR‐16 mimics decreased the expression of AKT3 and BCL2L2 and reduced the tumors volumes and weights, whereas the miR‐16 inhibitor exhibited adverse effects in the derived xenografts. In conclusion, the findings suggested that miR‐16 functions as a tumor suppressor miRNA to inhibit cell proliferation and induce apoptosis in OSCC through decreasing the oncogenes AKT3 and BCL2L2 and that miR‐16 could be a potential therapeutic target for OSCC. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9447-9457, December 2018.
  

  August 22, 2018   doi: 10.1002/jcp.26833   open full text
• Peroxidase expression is decreased by palmitate in cultured podocytes but increased in podocytes of advanced diabetic nephropathy.
  Eugene Lee, Hyun Soon Lee.
  Journal of Cellular Physiology. August 21, 2018
  --- - |2- High levels of serum free fatty acids (FFAs) are associated with lipotoxicity and type 2 diabetes. Palmitic acid (PA) is the predominant circulating saturated FFA. PA induces mitochondrial superoxide and hydrogen peroxide (H2O 2) generation in cultured podocytes. To elucidate the role of PA in antioxidant defense systems in diabetic nephropathy (DN), cultured podocytes were exposed to 250 μM PA for 1–24 hr, and protein expressions of catalase, peroxiredoxins (Prxs), and glutathione peroxidase (GPx) were examined by western blot analysis. PA induced an early transient increase in the Prx1, Prx2, and GPx1 levels in podocytes, but not catalase. Long‐term exposure of PA to podocytes significantly decreased the protein levels of Prx1, Prx2, GPx1, and catalase. Coincubation of PA‐treated cells with oleic acid, however, restored the expression of these proteins. In advanced human diabetic glomeruli, H2O2 generation was elevated as shown by increased fluorescence of dichlorofluorescein. Strong immunostaining for Prx1, Prx2, GPx1, and catalase was observed in the podocytes of advanced human DN, wherein transforming growth factor‐β1 staining was also positive. These results suggest that podocytes are susceptible to PA‐induced oxidative damage with impaired peroxidase activity and that peroxidases have futile antioxidant effects in the podocytes in the late stages of DN. Given this, PA‐induced podocyte injury via inadequate peroxidase response to H2O2 appears to play an important role in the pathogenesis of DN. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9060-9069, December 2018.
  

  August 21, 2018   doi: 10.1002/jcp.26875   open full text
• Methylation‐independent ITGA2 overexpression is associated with poor prognosis in de novo acute myeloid leukemia.
  Xin‐Yue Lian, Wei Zhang, De‐Hong Wu, Ji‐Chun Ma, Jing‐Dong Zhou, Zhi‐Hui Zhang, Xiang‐Mei Wen, Zi‐Jun Xu, Jiang Lin, Jun Qian.
  Journal of Cellular Physiology. August 21, 2018
  --- - |2- Previous studies have been indicated that integrin α2 (ITGA2) may be important in cell migration, invasion, survival, and angiogenesis. However, the correlation between ITGA2 expression and acute myeloid leukemia (AML) is still unclear. Real‐time quantitative polymerase chain reaction was carried out to analyze ITGA2 messenger RNA level. Methylation‐specific polymerase chain reaction (PCR) and bisulfite sequencing PCR were performed to detect the methylation of ITGA2 promoter. ITGA2 expression was significantly upregulated in 134 de novo AML patients compared with 33 controls (p = 0.007). ITGA2high group had markedly lower complete remission (CR) rate than ITGA2low group (p = 0.011). Furthermore, the overall survival in ITGA2high patients was significantly shorter than ITGA2low patients throughout AML cohort, non–acute promyelocytic leukemia (APL) and cytogenetic normal‐AML (p = 0.001, 0.002, and 0.044, respectively). Multivariate analysis confirmed that ITGA2 overexpression served as an independent prognostic factor in both whole‐cohort AML patients (p = 0.018) and non‐APL AML patients (p = 0.021). Besides, ITGA2 expression level was significantly decreased in AML patients after CR (p = 0.011), and was returned at the time of relapse phase (p = 0.021). Moreover, unmethylated ITGA2 promoter was identified in normal controls, leukemia cell lines, and primary leukemia cells with low or high ITGA2 expression. In conclusions, methylation‐independent ITGA2 overexpression is associated with poor prognosis in AML. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9584-9593, December 2018.
  

  August 21, 2018   doi: 10.1002/jcp.26866   open full text
• Peptidyl‐prolyl cis–trans isomerase NIMA interacting 1 regulates skeletal muscle fusion through structural modification of Smad3 in the linker region.
  Rabia Islam, Heein Yoon, Hye‐Rim Shin, Han‐Sol Bae, Bong‐Soo Kim, Won‐Joon Yoon, Kyung‐Mi Woo, Jeong‐Hwa Baek, Yun‐Sil Lee, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. August 21, 2018
  --- - |2- Myoblast fusion is critical for muscle growth, regeneration, and repair. We previously reported that the enzyme peptidyl‐prolyl cis–trans isomerase NIMA interacting 1 (Pin1) is involved in osteoclast fusion. The objective of this study was to investigate the possibility that Pin1 also inhibits myoblast fusion. Here, we show the increased number of nuclei in the Pin1+/− mice muscle fiber compared to that in wild‐type mice. Moreover, we show that low dose of the Pin1 inhibitor dipentamethylene thiuram monosulfide treatment caused enhanced fusion in C2C12 cells. The R‐Smads are well‐known mediators of muscle hypertrophy and hyperplasia as well as being substrates of Pin1. We found that Pin1 is crucial for maintaining the stability of Smad3 (homologues of the Drosophila protein, mothers against decapentaplegic (Mad) and the Caenorhabditis elegans protein Sma). Our results show that serine 204 within Smad3 is the key Pin1‐binding site during inhibition of myoblast fusion and that both the transforming growth factor‐β receptor and extracellular signal‐regulated kinase (ERK)‐mediated phosphorylation are required for the interaction of Pin1 with Smad3. These findings suggest that a precise level of Pin1 activity is essential for regulating myoblast fusion during myogenesis and muscle regeneration. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9390-9403, December 2018.
  

  August 21, 2018   doi: 10.1002/jcp.26774   open full text
• Inhibition of smoothened in breast cancer cells reduces CAXII expression and cell migration.
  Giuditta Guerrini, Mattia Criscuoli, Irene Filippi, Antonella Naldini, Fabio Carraro.
  Journal of Cellular Physiology. August 21, 2018
  --- - |2 Breast cancer (BC) relapse and metastasis are the leading cause of death and, together with drug resistance, keep mortality still high. The Hedgehog (Hh) pathway is expressed during embryogenesis, organogenesis and in adult tissue homeostasis and its aberrant activation is often associated with cancer. Carbonic anhydrase (CA) enzymes are important during development; they play a key role in controlling several cellular mechanisms, such as pH regulation, survival, and migration, and they are aberrantly expressed in cancer. The goal of this study was to investigate the interplay between the Hh pathway and CAXII in terms of BC cell migration. We here demonstrated that smoothened (SMO) silencing resulted in a reduction of CAXII expression at mRNA and protein level. This led to a decrease in cell migration, which was restored when cells were treated with an SMO agonist, Sag dihydrochloride (SAG), but not when cells were cotreated with SAG and the CAs inhibitor Acetazolamide. This suggested that the ability of SAG to promote cell migration was impaired when CAXII was inhibited. The reduction was also confirmed within hypoxic and inflammatory microenvironment, typical of BC, indicating a key role of the Hh pathway in controlling CAXII expression. Our results may contribute to further understand the physiology of BC cells and indicate that the Hh pathway controls BC cell migration and cell invasion also through CAXII, with important implications in identifying novel therapeutic targets. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9799-9811, December 2018.
  

  August 21, 2018   doi: 10.1002/jcp.26947   open full text
• Cancer metastasis versus stem cell homing: Role of platelets.
  Hojjat Naderi‐Meshkin, Naghmeh Ahmadiankia.
  Journal of Cellular Physiology. August 13, 2018
  --- - |2- Abstract One of the major obstacles in achieving a successful stem cell therapy is insufficient homing of transplanted cells. To overcome this obstacle, understanding the underlying mechanisms of stem cell homing is of obvious importance. Central to this review is the concept that cancer metastasis can be viewed as a role model to build up a comprehensive concept of stem cell homing. In this novel perspective, the prosurvival choices of the cancerous cells in the bloodstream, their arrest, extravasation, and proliferation at the secondary site can be exploited in favor of targeted stem cell homing. To date, tumor cells have been found to employ a wide variety of strategies to promote metastasis. One of these strategies is through their ability to activate platelets and subsequently activated platelets serve cancer cell survival and metastasis. Accordingly, in the first part of this review the roles of platelets in cancer metastasis as well as stem cell homing are discussed. Next, we provide some lessons learned from cancer metastasis in favor of developing strategies for improvement of stem cell homing with emphasis on the role of platelets. Based on direct or indirect evidence from metastasis, strategies such as manipulation of stem cells to enhance interaction with platelets, preconditioning–pretreatment of stem cells with platelets in vitro, and coinjection of both stem cells and platelets are proposed to improve stem cell homing. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9167-9178, December 2018.
  

  August 13, 2018   doi: 10.1002/jcp.26937   open full text
• Therapeutic potential of microRNAs in osteoporosis function by regulating the biology of cells related to bone homeostasis.
  Wenhua Zhao, Gengyang Shen, Hui Ren, De Liang, Xiang Yu, Zhida Zhang, Jinjing Huang, Ting Qiu, Jingjing Tang, Qi Shang, Peiyuan Yu, Zixian Wu, Xiaobing Jiang.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2- MicroRNAs (miRNAs) are novel regulatory factors that play important roles in numerous cellular processes through the posttranscriptional regulation of gene expression. Recently, deregulation of the miRNA‐mediated mechanism has emerged as an important pathological factor in osteoporosis. However, a detailed molecular mechanism between miRNAs and osteoporosis is still not available. In this review, the roles of miRNAs in the regulation of cells related to bone homeostasis as well as miRNAs that deregulate in human or animal are discussed. Moreover, the miRNAs that act as clusters in the biology of cells in the bone microenvironment and the difference of some important miRNAs for bone homeostasis between bone and other organs are mentioned. Overall, miRNAs that contribute to the pathogenesis of osteoporosis and their therapeutic potential are considered. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9191-9208, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26939   open full text
• Tumor‐associated macrophages and epithelial–mesenchymal transition in cancer: Nanotechnology comes into view.
  Roghayyeh Vakili‐Ghartavol, Reza Mombeiny, Arash Salmaninejad, Seyed Mahdi Rezayat Sorkhabadi, Reza Faridi‐Majidi, Mahmoud Reza Jaafari, Hamed Mirzaei.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2 Tumor‐associated macrophages (TAMs) are an important component of the leukocytic infiltrate of the tumor microenvironment. There is persuasive preclinical and clinical evidence that TAMs induce cancer inanition and malignant progression of primary tumors toward a metastatic state through a highly conserved and fundamental process known as epithelial–mesenchymal transition (EMT). Tumor cells undergoing EMT are distinguished by increased motility and invasiveness, which enable them to spread to distant sites and form metastases. In addition, besides becoming resistant to apoptosis and antitumor drugs, they also contribute to immunosuppression and get a cancer stem‐cell like phenotype. Here, we will focus on selected molecular pathways underlying EMT—in particular, the role of TAMs in the induction and maintenance of EMT—and further discuss how the targeting of TAMs through the application of nanotechnology tools allows the development of a whole new range of therapeutics. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9223-9236, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.27027   open full text
• The MicroRNA‐326: Autoimmune diseases, diagnostic biomarker, and therapeutic target.
  Golamreza Jadideslam, Khalil Ansarin, Ebrahim Sakhinia, Shahriar Alipour, Farhad Pouremamali, Alireza Khabbazi.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2- MicroRNAs (miRNAs) are uniquely regulated in healthy, inflamed, activated, cancerous, or other cells and tissues of a pathological state. Many studies confirm that immune dysregulation and autoimmune diseases with inflammation are correlated with various miRNA expression changes in targeted tissues and cells in innate or adaptive immunity. In this review, we will explain the history and classification of epigenetic changes. Next, we will describe the role of miRNAs changes, especially mir‐326 in autoimmunity, autoinflammatory, and other pathological conditions. A systematic search of MEDLINE, Embase, and Cochrane Library was presented for all related studies from 1899 to 2017 with restrictions in the English language. In recent years, researchers have concentrated on mostly those roles of miRNA that are correlated with the inflammatory and anti‐inflammatory process. Latest studies have proposed a fundamental pathogenic role in cancers and autoinflammatory diseases. Studies have described the role of microRNAs in autoimmunity and autoinflammatory diseases, cancers, and so on. The miRNA‐326 expression plays a significant role in autoimmune and other types of diseases. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9209-9222, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26949   open full text
• Cellular senescence: Molecular mechanisms and pathogenicity.
  Wenqiang Wei, Shaoping Ji.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2- Cellular senescence is the arrest of normal cell division. Oncogenic genes and oxidative stress, which cause genomic DNA damage and generation of reactive oxygen species, lead to cellular senescence. The senescence‐associated secretory phenotype is a distinct feature of senescence. Senescence is normally involved in the embryonic development. Senescent cells can communicate with immune cells to invoke an immune response. Senescence emerges during the aging process in several tissues and organs. In fact, increasing evidence shows that cellular senescence is implicated in aging‐related diseases, such as nonalcoholic fatty liver disease, obesity and diabetes, pulmonary hypertension, and tumorigenesis. Cellular senescence can also be induced by microbial infection. During cellular senescence, several signaling pathways, including those of p53, nuclear factor‐κB (NF‐κB), mammalian target of rapamycin, and transforming growth factor‐beta, play important roles. Accumulation of senescent cells can trigger chronic inflammation, which may contribute to the pathological changes in the elderly. Given the variety of deleterious effects caused by cellular senescence in humans, strategies have been proposed to control senescence. In this review, we will focus on recent studies to provide a brief introduction to cellular senescence, including associated signaling pathways and pathology. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9121-9135, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26956   open full text
• Hyaluronidase2 (Hyal2) modulates low shear stress‐induced glycocalyx impairment via the LKB1/AMPK/NADPH oxidase‐dependent pathway.
  Hongfeng Yang, Linlin Zhu, Yuelin Chao, Yue Gu, Xiangquan Kong, Mingxing Chen, Peng Ye, Jie Luo, Shaoliang Chen.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2- The endothelium glycocalyx layer (ECL), presents on the apical surface of endothelial cells, creates a barrier between circulating blood and the vessel wall. Low shear stress (LSS) may accelerate the degradation of the glycocalyx via hyaluronidase2 (Hyal2) and then alter the cell polarity. Yet the liver kinase B1 (LKB1) signaling pathway plays an important role in regulating cell polarity. However, the relationship between LKB1 and glycocalyx during LSS is not clear. In the current study, we demonstrate that LSS attenuates LKB1 and AMP‐activated protein kinase activation as well as activated nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (p47phox) and Hyal2 in the human umbilical vein endothelial cell (HUVEC). Pretreatment with 5‐Aminoimidazole‐4‐carboxamide1‐β‐D‐ribofuranoside (AICAR), or diphenyleneiodonium (DPI chloride) and transfection with LKB1 overexpression vector and p47phox small interfering RNA downregulated LSS‐induced Hyal2 activation. By coimmunoprecipitation, we discovered the existence of p47phox/Hyal2 complex. LSS induced the dissociation of p47phox/Hyal2 complex, which was inhibited by LKB1 overexpression and AICAR. Furthermore, knockdown of Hyal2 performed a positive feedback on LKB1 activity. In addition, we also show that LSS enhanced LKB1 translocation from the cytosol to the nucleus. Taken together, these data indicate that Hyal2 regulates LSS‐induced injury of the glycocalyx via LKB1/AMPK/NADPH oxidase signaling cascades. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9701-9715, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26944   open full text
• Identification of key genes and pathways associated with osteogenic differentiation of adipose stem cells.
  Xinyuan Zhao, Minlu Liang, Xiaona Li, Xiaoling Qiu, Li Cui.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2 Adipose stem cells (ASCs) are considered a great alternative source of mesenchymal stem cells (MSCs) and have shown great promise on tissue engineering and regenerative medicine applications, including bone repair. However, the underlying mechanisms regulating the osteogenic differentiation of ASCs remain poorly known. Gene expression profiles of GSE63754 and GSE37329 were downloaded from gene expression omnibus database. R software and Bioconductor packages were used to compare and identify the differentially expressed genes (DEGs) before and after ASC osteogenic differentiation. The common significant DEGs between GSE63754 and GSE37329 were then subjected to gene ontology (GO) enrichment analysis, ingenuity pathway analysis (IPA), and protein–protein interactions (PPIs) networks analysis. One of the central node genes FOXO1 was selected for further investigation. A total of 142 up‐ and 69 downregulated genes were aberrantly expressed in both GSE63754 and GSE37329. GO analysis revealed that these DEGs were associated with extracellular matrix organization, proteinaceous extracellular matrix, and Wnt‐protein binding. IPA analysis showed that canonical pathways, such as FXR/RXR activation, adipogenesis pathway, and LXR/RXR activation, were involved in regulating osteogenic differentiation of ASCs. A total of three subnetworks and 39 nodes were identified with PPI network and MCODE plugin. Moreover, suppression of one central node gene FOXO1 inhibited the osteogenic differentiation of ASCs. Our study provides a registry of genes and pathways that play important roles in regulating osteogenic differentiation of ASCs, which might have potential therapeutic applications in bone regeneration and bone tissue engineering. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9777-9785, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26943   open full text
• Establishment and characterization of a telomerase‐immortalized porcine bronchial epithelial cell line.
  Xing Xie, Yuan Gan, Maoda Pang, Guoqing Shao, Lei Zhang, Beibei Liu, Qi Xu, Haiyan Wang, Yanyan Feng, Yanfei Yu, Rong Chen, Meng Wu, Zhenzhen Zhang, Lizhong Hua, Qiyan Xiong, Maojun Liu, Zhixin Feng.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2 Primary porcine bronchial epithelial cells (PBECs) are an ideal model to study the molecular and pathogenic mechanisms of various porcine respiratory pathogens. However, the short lifespan of primary PBECs greatly limit their application. Here, we isolated and cultured primary PBECs and established immortalized PBECs by transfecting primary PBECs with the pEGFP‐hTERT recombinant plasmid containing human telomerase reverse transcriptase (hTERT). Immortalized PBECs (hTERT‐PBECs) retained the morphological and functional features of primary PBECs as indicated by cytokeratin 18 expression, telomerase activity assay, proliferation assays, karyotype analysis, and quantitative reverse‐transcriptase polymerase chain reaction. Compared to primary PBECs, hTERT‐PBECs had higher telomerase activity, extended replicative lifespan, and displayed enhanced proliferative activity. Moreover, this cell line is not transformed in vitro and does not exhibit a malignant phenotype in vivo, suggesting that it can be safely used in further studies. Besides, hTERT‐PBECs were susceptible to swine influenza virus of H3N2 subtype and porcine circovirus type 2. In conclusion, the immortalized hTERT‐PBECs represent a valuable in vitro model, which can be widely used in the study of porcine respiratory pathogenic infections. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9763-9776, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26942   open full text
• Exogenous hydrogen sulfide attenuates the development of diabetic cardiomyopathy via the FoxO1 pathway.
  Peng Ye, Yue Gu, Yan‐Rong Zhu, Yue‐Lin Chao, Xiang‐Quan Kong, Jie Luo, Xiao‐Min Ren, Guang‐Feng Zuo, Dai‐Min Zhang, Shao‐Liang Chen.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2+ Background Previous studies have suggested that exogenous hydrogen sulfide can alleviate the development of diabetic cardiomyopathy (DCM) by inhibiting oxidative stress, inflammation, and apoptosis. However, the underlying mechanism is not fully understood. Nuclear expression and function of the transcription factor Forkhead box protein O (FoxO1) have been associated with cardiovascular diseases, and thus, the importance of FoxO1 in DCM has gained increasing attention. This study was designed to investigate the interactions between hydrogen sulfide (H2S) and nuclear FoxO1 in DCM. Methods Diabetes was induced in adult male C57BL/6J mice by intraperitoneal injection of streptozotocin and was treated with H2S donor sodium hydrosulfide for 12 weeks. The H9C2 cardiomyoblast cell line and neonatal rat cardiomyocytes (NRCMs) were treated with the slow‐releasing H2S donor GYY4137 before high‐glucose (HG) exposure with or without pretreatment with the Akt inhibitor MK‐2206 2HCl. Changes in FoxO1 protein phosphorylation and subcellular localization were determined in H9C2 cells, NRCMs, and cardiac tissues from normal and diabetic mice. Cardiac structure and function in the diabetic mice were evaluated by echocardiography and histological analysis and compared with those in control animals. Results The echocardiographic and histopathological data indicated that exogenous H2S improved cardiac function and attenuated cardiac hypertrophy and myocardial fibrosis in diabetic mice. H2S also improved HG‐induced oxidative stress and apoptosis in cardiac tissue and NRCMs. In addition, H2S induced FoxO1 phosphorylation and nuclear exclusion in vitro and in vivo, and this function was not inhibited by MK‐2206 2HCl. Alanine substitution mutation of three sites in FoxO1‐enhanced FoxO1 transcriptional activity, and subsequent treatment with exogenous H2S could not prevent HG‐induced nuclear retention. Conclusions Our data indicate that H2S is a novel regulator of FoxO1 in cardiac cells and provide evidence supporting the potential of H2S in inhibiting the progression of DCM. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9786-9798, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26946   open full text
• Berberine attenuates pulmonary arterial hypertension via protein phosphatase 2A signaling pathway both in vivo and in vitro.
  Jie Luo, Yue Gu, Pengfei Liu, Xiaomin Jiang, Wande Yu, Peng Ye, Yuelin Chao, Hongfeng Yang, Linlin Zhu, Ling Zhou, Shaoliang Chen.
  Journal of Cellular Physiology. August 05, 2018
  --- - |2- Excessive proliferation, migration, and antiapoptosis of pulmonary artery (PA) smooth muscle cells (PASMCs) underlies the development of pulmonary vascular remodeling. The innervation of the PA is predominantly sympathetic, and increased levels of circulating catecholamines have been detected in pulmonary arterial hypertension (PAH), suggesting that neurotransmitters released by sympathetic overactivation may play an essential role in PAH. However, the responsible mechanism remains unclear. Here, to investigate the effects of norepinephrine (NE) on PASMCs and the related mechanism, we used 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide, the proliferating cell nuclear antigen and the cell counting kit‐8 assay to evaluate the proliferation of PASMCs, Boyden chamber migration, and wound‐healing assays to assess migration and western blot analysis to investigate protein expression. We demonstrated that the phosphorylation level of the protein phosphatase 2A (PP2A) catalytic subunit (Y307) was higher in PAH patients and PAH models than in controls, both in vivo and in vitro. In addition, NE induced the proliferation and migration of PASMCs, which was attenuated by berberine (BBR), a Chinese herbal medicine, and/or PP2A overexpression. PP2A inhibition worsened NE‐induced PAH and could not be reversed by BBR. Thus, PP2A is critical in driving PAH, and BBR may alleviate PAH via PP2A signaling pathways, thereby offering a potential therapeutic option for PAH. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9750-9762, December 2018.
  

  August 05, 2018   doi: 10.1002/jcp.26940   open full text
• Infection‐associated epigenetic alterations in gastric cancer: New insight in cancer therapy.
  Sadegh Fattahi, Mohadeseh Kosari‐Monfared, Elham Ghadami, Monireh Golpour, Parastoo Khodadadi, Mohammad Ghasemiyan, Haleh Akhavan‐Niaki.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- Gastric cancer risk is higher for malignancies motivated by bacterial and viral infections. Epigenetic abnormalities including DNA methylation, histone modifications, and noncoding RNAs are important regulatory key players in gastric cancer development in infected patients. Epigenetic memory restoration is an extremely interesting phenomenon which should be considered in therapeutic approaches. In vitro and in vivo antiviral treatments in combination with epigenetic therapeutic strategies along with standard chemotherapy revealed promising outcomes in gastric cancer prevention and treatment. This review summarizes our current understanding of the gastric cancer infections and epigenetic alterations caused by these agents. We focus on studies highlighting recent advances in epigenetic restoration by target specific drugs and present also a comprehensive overview of effective antiviral drug treatments against gastric cancer. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9261-9270, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.27030   open full text
• Ferritinophagy/ferroptosis: Iron‐related newcomers in human diseases.
  Mingzhu Tang, Zhe Chen, Di Wu, Linxi Chen.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- Nuclear receptor coactivator 4 mediated ferritinophagy is an autophagic phenomenon that specifically involves ferritin to release intracellular free iron. Ferritinophagy is implicated in maintaining efficient erythropoiesis. Notably, ferritinophagy also plays a central role in driving some pathological processes, including Parkinson’s disease (PD) and urinary tract infections. Some evidence has demonstrated that ferritinophagy is critical to induce ferroptosis. Ferroptosis is a newly nonapoptotic form of cell death, characterized by the accumulation of iron‐based lipid reactive oxygen species. Ferroptosis plays an important role in inhibiting some types of cancers, such as hepatocellular carcinoma, pancreatic carcinoma, prostate cancer, and breast cancer. Conversely, the activation of ferroptosis accelerates neurodegeneration diseases, including PD and Alzheimer’s disease. Therefore, in this review, we summarize the regulatory mechanisms related to ferritinophagy and ferroptosis. Moreover, the distinctive effects of ferritinophagy in human erythropoiesis and some pathologies, coupled with the promotive or inhibitory role of tumorous and neurodegenerative diseases mediated by ferroptosis, are elucidated. Obviously, activating or inhibiting ferroptosis could be exploited to achieve desirable therapeutic effects on diverse cancers and neurodegeneration diseases. Interrupting ferritinophagy to control iron level might provide a potentially therapeutic avenue to suppress urinary tract infections. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9179-9190, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.26954   open full text
• Demethoxycurcumin: A naturally occurring curcumin analogue with antitumor properties.
  Mahdi Hatamipour, Mahin Ramezani, Sayyed Abolghasem Sajadi Tabassi, Thomas P. Johnston, Mahnaz Ramezani, Amirhosein Sahebkar.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- The eradication of cancer in a patient remains an elusive challenge despite advances in early detection and diagnosis, chemo‐ and immunotherapy, pinpoint radiation treatments, and expert surgical intervention. Although significant gains have been made in our understanding of cancer cell biology, a definite cure for most cancers does not exist at present. Thus, it is not surprising that the research and medical communities continue to explore the importance and therapeutic potential of natural products in their multimodality cancer treatment approach. Curcuminoids found in turmeric are one such class of natural products that have been extensively investigated for their potential to halt the progression of cancer cell proliferation and, more important, to stop metastasis from occurring. In this review, we examine one curcuminoid (demethoxycurcumin [DMC]) largely because of its increased stability and better aqueous solubility at physiological pH, unlike the more well‐known curcuminoid (curcumin), which is largely unabsorbed after oral ingestion. The present review will focus on the signaling pathways that DMC utilizes to modulate the growth, invasion, and metastasis of cancer cells in an effort to provide enhanced mechanistic insight into DMC’s action as it pertains to brain, ovarian, breast, lung, skin, and prostate cancer. Additionally, this review will attempt to provide an overview of DMC’s mechanism of action by modulating apoptosis, cell cycle, angiogenesis, metastasis, and chemosensitivity. Lastly, it is hoped that increased understanding will be gained concerning DMC’s interactive role with microRNA‐551a, 5′ adenosine monophosphate‐activated protein kinase, nuclear factor‐κB, Wnt inhibitory factor‐1, and heat shock protein 70 to affect the progression of cancer. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9247-9260, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.27029   open full text
• Monocyte‐to‐HDL‐cholesterol ratio as a prognostic marker in cardiovascular diseases.
  Shiva Ganjali, Antonio M. Gotto, Massimiliano Ruscica, Stephen L. Atkin, Alexandra E. Butler, Maciej Banach, Amirhossein Sahebkar.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- Inflammation and lipid accumulation are two basic hallmarks of atherosclerosis as a chronic disease. Inflammation not only is a local response but can also be considered as a systemic process followed by an elevation of inflammatory mediators. Monocytes are a major source of proinflammatory species during atherogenesis. In atherosclerosis, modified low‐density lipoproteins (LDLs) are removed by macrophages; these are recruited in the vessel wall, inducing the release of inflammatory cytokines in inflamed tissue. Hence, inflammatory cholesterol ester‐loaded plaque is generated. High‐density lipoprotein‐cholesterol (HDL‐C) exhibits antiatherosclerotic effects by neutralizing the proinflammatory and pro‐oxidant effects of monocytes via inhibiting the migration of macrophages and LDL oxidation in addition to the efflux of cholesterol from these cells. Furthermore, HDL plays a role in suppressing the activation of monocytes and proliferation–differentiation of monocyte progenitor cells. Thus, accumulation of monocytes and reduction of HDL‐C may participate in atherosclerosis and cardiovascular diseases (CVD). Given that the relationship between the high number of monocytes and low HDL‐C levels has been reported in inflammatory disorders, this review focused on understanding whether the monocyte‐to‐HDL ratio could be a convenient marker to predict atherosclerosis development and progression, hallmarks of CV events, instead of the individual monocyte count or HDL‐C level. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9237-9246, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.27028   open full text
• The role of ErbB3 binding protein 1 in cancer: Friend or foe?
  Dang Quan Nguyen, Dinh Hoa Hoang, Thanh Thao Nguyen Vo, Vu Huynh, Lucy Ghoda, Guido Marcucci, Le Xuan Truong Nguyen.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- ErbB3, a member of the epidermal growth factor receptor family, reportedly plays an essential role in the regulation of cancer progression and therapeutic resistance. Numerous studies have indicated that ErbB3 binding protein 1 (Ebp1), a binding partner for ErbB3, plays an important regulatory role in the expression and function of ErbB3, but there is no agreement as to whether Ebp1 also has an ErbB3‐independent function in cancer and how it might contribute to tumorigenesis. In this review, we will discuss the different functions of the two Ebp1 isoforms, p48 and p42, that may be responsible for the potentially dual role of Ebp1 in cancer growth. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9110-9120, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.26951   open full text
• Analysis of pro‐ and anti‐inflammatory cytokine gene variants and serum cytokine levels as prognostic markers in breast cancer.
  Raman Preet Kaur, Kanika Vasudeva, Heena Singla, Raja Paramjeet Singh Benipal, Preeti Khetarpal, Anjana Munshi.
  Journal of Cellular Physiology. August 04, 2018
  --- - |2- The aim of current study was to evaluate the genetic variation in all the genes encoding pro‐ and anti‐inflammatory cytokines in association with breast cancer development in patients from Malwa region of Punjab. The importance of the levels of interleukin (IL)‐17, tumor necrosis factor, interferon γ, IL‐10, IL‐6, IL‐4, and IL‐2 with respect to clinicopathological data, prognosis, and disease‐free survival was also determined in these patients. Two hundred and fifty female breast cancer patients and 250 age‐matched controls were screened for variations in cytokine‐encoding genes using global screening array microchip and PCR‐RFLP. The level of cytokines was estimated in 150 patients and 60 age‐matched controls using BD™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 cytokine kit by BD Accuri flow cytometer. The difference in cytokine levels was evaluated by Mann–Whitney test. No significant variation in the genes encoding various cytokines was found between patients and controls. Out of the seven cytokines evaluated, the levels of IL‐6 and IL‐17a were found to be significantly high in patients in comparison with controls ( p = 0.001 and 0.02, respectively). The elevated levels of these cytokines are also associated significantly with poor outcome. We did not find any specific variation in the genes encoding various cytokines between patients and controls. However, there was a significant difference in the serum levels of IL‐6 and IL‐17a between patients and controls, and the elevated levels of these two cytokines associated significantly with poor outcome in breast cancer patients and, therefore, can be used as prognostic markers. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9716-9723, December 2018.
  

  August 04, 2018   doi: 10.1002/jcp.26901   open full text
• STAC3 incorporation into skeletal muscle triads occurs independent of the dihydropyridine receptor.
  Marta Campiglio, Mehmet M. Kaplan, Bernhard E. Flucher.
  Journal of Cellular Physiology. August 02, 2018
  --- - |2- Excitation‐contraction (EC) coupling in skeletal muscles operates through a physical interaction between the dihydropyridine receptor (DHPR), acting as a voltage sensor, and the ryanodine receptor (RyR1), acting as a calcium release channel. Recently, the adaptor protein SH3 and cysteine‐rich containing protein 3 (STAC3) has been identified as a myopathy disease gene and as an additional essential EC coupling component. STAC3 interacts with DHPR sequences including the critical EC coupling domain and has been proposed to function in linking the DHPR and RyR1. However, we and others demonstrated that incorporation of recombinant STAC3 into skeletal muscle triads critically depends only on the DHPR but not the RyR1. On the contrary, here, we provide evidence that endogenous STAC3 incorporates into triads in the absence of the DHPR in myotubes and muscle fibers of dysgenic mice. This finding demonstrates that STAC3 interacts with additional triad proteins and is consistent with its proposed role in directly or indirectly linking the DHPR with the RyR1. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9045-9051, December 2018.
  

  August 02, 2018   doi: 10.1002/jcp.26767   open full text
• Exercise activates the hypothalamic S1PR1–STAT3 axis through the central action of interleukin 6 in mice.
  Vagner R. R. Silva, Thayana O. Micheletti, Carlos K. Katashima, Luciene Lenhare, Joseane Morari, Alexandre Moura‐Assis, José C. Lima‐Júnior, Juliana A. Camargo, Gabriela R. Passos, Rodrigo S. Gaspar, Licio A. Velloso, Mario J. Saad, Adelino S. R. da Silva, Leandro P. Moura, Dennys E. Cintra, José R. Pauli, Eduardo R. Ropelle.
  Journal of Cellular Physiology. July 31, 2018
  --- - |2 Hypothalamic sphingosine‐1‐phosphate receptor 1 (S1PR1), the G protein–coupled receptor 1 of sphingosine‐1‐phosphate, has been described as a modulator in the control of energy homeostasis in rodents. However, this mechanism is still unclear. Here, we evaluate the role of interleukin 6 (IL‐6) associated with acute physical exercise in the control of the hypothalamic S1PR1–signal transducer and activator of transcription 3 (STAT3) axis. Acute exercise session and an intracerebroventricular IL‐6 injection increased S1PR1 protein content and STAT3 phosphorylation in the hypothalamus of lean and obese mice accompanied by a reduction in food consumption. Transcriptome analysis indicated a strong positive correlation between Il‐6 and S1pr1 messenger RNA in several tissues of genetically diverse BXD mice strains and humans, including in the hypothalamus. Interestingly, exercise failed to stimulate the S1PR1–STAT3 axis in IL‐6 knockout mice and the disruption of hypothalamic‐specific IL‐6 action blocked the anorexigenic effects of exercise. Taken together, our results indicate that physical exercise modulates the S1PR1 protein content in the hypothalamus, through the central action of IL‐6. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9426-9436, December 2018.
  

  July 31, 2018   doi: 10.1002/jcp.26818   open full text
• Small molecule inhibitor RepSox prevented ovariectomy‐induced osteoporosis by suppressing osteoclast differentiation and bone resorption.
  Liangwei Mei, Wenhua Sang, Zhenzhong Chen, Lin Zheng, Kangtao Jin, Chao Lou, Wenjun Huang, Dengwei He.
  Journal of Cellular Physiology. July 30, 2018
  --- - |2- Osteoporosis (OP) is a serious metabolic disease that, due to the increased number or function of osteoclasts, results in increased bone brittleness and, therefore, fragile fracture. Some recent studies report the importance of the transforming growth factor β (TGFβ) pathway in bone homeostasis. RepSox is a small molecule inhibitor of TGFβRI that has a wide range of potential application in clinical medicine, except OP. The aim of our study is to evaluate the effects of RepSox on the differentiation and bone resorption of osteoclasts in vitro and in vivo in an ovariectomy (OVX)‐induced OP model. An initial analysis showed TGFβRI messenger RNA expression in both bone samples and bone cells. In the in vitro study, RepSox inhibited the receptor activator of nuclear factor κB ligand (RANKL)‐induced osteoclast differentiation and bone resorption activity. Real‐time polymerase chain reaction (PCR) analysis showed that RepSox suppressed osteoclastic marker gene expression in both dose‐dependent and time‐dependent manners. In addition, RepSox did not affect osteoblast differentiation, migration or osteoblastic‐specific gene expression in vitro. Furthermore, western blot analysis indicated the underlying mechanisms of the RepSox suppression of osteoclastogenesis via the Smad3 and c‐Jun N‐terminal kinase/activator protein‐1 (JNK/AP‐1) signaling pathways. Finally, our animal experiments revealed that RepSox prevented OVX‐induced bone loss in vivo. Together, our data suggest that RepSox regulates osteoclast differentiation, bone resorption, and OVX‐induced OP via the suppression of the Smad3 and JNK/AP‐1 pathways. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9724-9738, December 2018.
  

  July 30, 2018   doi: 10.1002/jcp.26914   open full text
• Deficiency of fibroblast growth factor 2 (FGF‐2) leads to abnormal spermatogenesis and altered sperm physiology.
  Lucía Saucedo, Regina Rumpel, Cristian Sobarzo, Dietmar Schreiner, Gudrun Brandes, Livia Lustig, Mónica Hebe Vazquez‐Levin, Claudia Grothe, Clara Marín‐Briggiler.
  Journal of Cellular Physiology. July 27, 2018
  --- - |2- In previous studies, we described the presence of fibroblast growth factor 2 (FGF‐2) and its receptors (FGFRs) in human testis and sperm, which are involved in spermatogenesis and in motility regulation. The aim of the present study was to analyze the role of FGF‐2 in the maintenance of sperm physiology using FGF‐2 knockout (KO) mice. Our results showed that in wild‐type (WT) animals, FGF‐2 is expressed in germ cells of the seminiferous epithelium, in epithelial cells of the epididymis, and in the flagellum and acrosomal region of epididymal sperm. In the FGF‐2 KO mice, we found alterations in spermatogenesis kinetics, higher numbers of spermatids per testis, and enhanced daily sperm production compared with the WT males. No difference in the percentage of sperm motility was detected, but a significant increase in sperm concentration and in sperm head abnormalities was observed in FGF‐2 KO animals. Sperm from KO mice depicted reduced phosphorylation on tyrosine residues (a phenomenon that was associated with sperm capacitation) and increased acrosomal loss after incubation under capacitating conditions. However, the FGF‐2 KO males displayed no apparent fertility defects, since their mating with WT females showed no differences in the time to delivery, litter size, and pup weight in comparison with WT males. Overall, our findings suggest that FGF‐2 exerts a role in mammalian spermatogenesis and that the lack of FGF‐2 leads to dysregulated sperm production and altered sperm morphology and function. FGF‐2‐deficient mice constitute a model for the study of the complex mechanisms underlying mammalian spermatogenesis. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9640-9651, December 2018.
  

  July 27, 2018   doi: 10.1002/jcp.26876   open full text
• CerS6 regulates cisplatin resistance in oral squamous cell carcinoma by altering mitochondrial fission and autophagy.
  Shengjiao Li, Yangou Wu, Yunpeng Ding, Miao Yu, Zexin Ai.
  Journal of Cellular Physiology. July 27, 2018
  --- - |2 Chemoresistance remains a challenge in the effective treatment of solid tumors, including oral squamous cell carcinoma (OSCC). Mitochondrial dynamics and autophagy have recently been implicated in the chemoresistance of cancer cells. The neutralization of ceramide is also associated with multidrug resistance, and ceramide synthase 6 (CerS6) is known to induce apoptosis. However, whether CerS6 regulates chemoresistance in OSCC is not clearly understood. Therefore, we investigated the role of CerS6 in the susceptibility of OSCC cells to cisplatin. In this study, we observed that cisplatin‐resistant OSCC cells process lower levels of fission‐state mitochondria and cell apoptosis than cisplatin‐sensitive cells, and autophagy was activated in cisplatin‐resistant OSCC cells. We found lower CerS6 expression in cisplatin‐resistant OSCC cells. Overexpression of CerS6 with lentivirus‐encoded CerS6 complementary DNA in cisplatin‐resistant OSCC cells increased cisplatin sensitivity. Overexpression of CerS6 enhanced mitochondrial fission and apoptosis and attenuated cisplatin‐induced autophagy in cisplatin‐resistant OSCC cells. Further investigation indicated that CerS6 might function through altering calpain expression to enhance cisplatin sensitivity. Cisplatin‐resistant OSCC cells xenografted onto a nude mouse model confirmed that CerS6 enhanced cisplatin chemotherapy sensitivity to reduce tumor volume. These data indicate that CerS6 could mediate an effective response to cisplatin in chemoresistant OSCC. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9416-9425, December 2018.
  

  July 27, 2018   doi: 10.1002/jcp.26815   open full text
• Effect of microRNA‐186 on oxidative stress injury of neuron by targeting interleukin 2 through the janus kinase‐signal transducer and activator of transcription pathway in a rat model of Alzheimer’s disease.
  Dong‐Mei Wu, Xin Wen, Yong‐Jian Wang, Xin‐Rui Han, Shan Wang, Min Shen, Shao‐Hua Fan, Juan Zhuang, Zi‐Feng Zhang, Qun Shan, Meng‐Qiu Li, Bin Hu, Chun‐Hui Sun, Jun Lu, Gui‐Quan Chen, Yuan‐Lin Zheng.
  Journal of Cellular Physiology. July 11, 2018
  --- - |2 Recent studies have proposed that microRNAs (miR) function as novel diagnostic and prognostic biomarkers and therapeutic targets in Alzheimer’s disease (AD), a common disease among the elderly. In the current study, we aim to explore the effect of miR‐186 on oxidative stress injury of neuron in rat models of AD with the involvement of the interleukin‐2 (IL2) and the Janus kinase/signal transducers and activators of transcription (JAK–STAT) pathways. AD rat models were established, and dual‐luciferase reporter assay and online software were used to confirm the targeting relationship between miR‐186 and IL2. Immunohistochemistry was used evaluating the positive rate of IL2. Afterward, to define the role of miR‐186 in AD, miR‐186, IL2, and JAK–STAT related protein (JAK2, STAT3) expressions were quantified. Cell proliferation was measured by 3‐(4,5‐dimethylthiazol‐2‐yl)2,5‐diphenyl tetrazolium bromide, and cell apoptosis was detected by flow cytometry. We observed downregulated miR‐186 and IL2 and upregulated JAK–STAT signaling pathway related genes in AD. The overexpression of miR‐186 was shown to significantly promote cell proliferation while suppressing cell apoptosis along with the expression of the IL2 and JAK–STAT signaling pathway related protein. Collectively, the key findings obtained from the current study define the potential role of miR‐186 as an inhibitor of AD development by downregulation of IL2 through suppression of the JAK–STAT signaling pathway. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9488-9502, December 2018.
  

  July 11, 2018   doi: 10.1002/jcp.26843   open full text
• Differentiated adipose‐derived stem cell cocultures for bone regeneration in RADA16‐I in vitro.
  Huifang Yang, Nanrui Hong, Hsiaowei Liu, Jieda Wang, Yan Li, Shuyi Wu.
  Journal of Cellular Physiology. July 11, 2018
  --- - |2- Craniofacial defects can cause morbidness. Adipose‐derived stem cells (ADSCs) have shown great promise for osteogeneration and vascularization; therefore cocultures of differentiated ADSCs are explored to increase bone and vessel formation. In this study, ADSCs were induced into osteogenic ADSCs (os‐ADSCs) and endothelial ADSCs (endo‐ADSCs) cells, which were then cocultured in variable proportions (os‐ADSCs/endo‐ADSCs = 2:1, 1:1, 1:2). The os‐ADSCs in a ratio of 1:1 expressed more ALP, RUNX2 and COL‐I, whereas VEGF, vWF and CD31 were upregulated in the endo‐ADSCs of this group. Next generation RNA sequencing (RNA‐seq) was performed to evaluate the molecular mechanisms of cocultured ADSCs. The os‐ADSCs and endo‐ADSCs interacted with each other during osteogenic and angiogenic differentiation, especially at the ratio of 1:1, and were regulated by vascular‐related genes, cell‐mediated genes, bone‐related genes and the transforming growth factor β signaling pathway (TGF‐β), mitogen‐activated protein kinase signaling pathway (MAPK) and wnt signaling pathway (Wnt). Angptl4, apoe, mmp3, bmp6, mmp13 and fgf18 were detected to be up‐regulated, and cxcl12 and wnt5a were down‐regulated. The results showed that the gene expression levels were consistent with that in RNA‐seq. The cells were then seeded into self‐assembling peptide RADA16‐I scaffolds as cocultures (1:1) and monocultures (ADSCs, os‐ADSCs, endo‐ADSCs). The results showed that the cells of all groups grew and proliferated well on the scaffolds, and the cocultured group exhibited better osteogeneration and vascularization. In conclusion, cocultured os‐ADSCs and endo‐ADSCs at the ratio of 1:1 showed strong osteogenic and angiogenic differentiation. There is a great potential for osteogenesis and vascularization by 3D culturing cells in a 1:1 ratio in self‐assembling peptide RADA16‐I scaffolds, which requires evaluation for bone regeneration in vivo. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9458-9472, December 2018.
  

  July 11, 2018   doi: 10.1002/jcp.26838   open full text
• Stable overexpression of p130/E2F4 affects the multipotential abilities of bone‐marrow‐derived mesenchymal stem cells.
  Xiwen Zhang, Jianxiao Chen, Airan Liu, Xiuping Xu, Ming Xue, Jingyuan Xu, Yi Yang, Haibo Qiu, Fengmei Guo.
  Journal of Cellular Physiology. July 10, 2018
  --- - |2- Bone‐marrow‐derived mesenchymal stem cells (MSCs) have great potential in transplantation medicine due to their multiple advantages. However, the controlled differentiation of MSCs is one of the key aspects of effective clinical transplantation. Growing evidence suggests that the cell cycle plays an important role in regulating differentiation, while p130 and E2F4 are key to cell cycle checkpoints. The aim of the study is to evaluate the effects and mechanism of p130/E2F4 on the multidifferentiation of MSCs. Our data showed that the transduction efficiencies of p130 or E2F4 mediated by lentiviral vectors were 80.3%–84.4%. p130 and E2F4 mRNA expression was significantly higher in MSC‐p130 and MSC‐E2F4 cells than in MSC normal control (NC) cells. Similar results were also observed for p130 and E2F4 protein expression. After osteogenic or adipogenic differentiation, the G1 phase was significantly delayed in the MSC‐p130 and MSC‐E2F4 groups compared with that in the MSC‐NC group. However, the G1 phase in the MSC‐p130 and MSC‐E2F4 groups did the opposite after chondrogenic differentiation. Moreover, overexpressing p130 or E2F4 significantly improved osteogenic differentiation while inhibiting adipogenic and chondrogenic differentiation of mouse MSCs (mMSCs). Moreover, overexpressing p130 or E2F4 significantly improved migration but not proliferation of mMSCs. Our data suggest that cell cycle regulation may be involved in p130/E2F4‐mediated changes in the multipotential abilities of bone‐marrow‐derived mMSCs. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9739-9749, December 2018.
  

  July 10, 2018   doi: 10.1002/jcp.26926   open full text
• Epithelial‐to‐mesenchymal transition and cancer stem cells contribute to breast cancer heterogeneity.
  Deli Hong, Andrew J. Fritz, Sayyed K. Zaidi, Andre J. Wijnen, Jeffrey A. Nickerson, Anthony N. Imbalzano, Jane B. Lian, Janet L. Stein, Gary S. Stein.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 Breast cancer is the most common cancer in women, and accounts for ~30% of new cancer cases and 15% of cancer‐related deaths. Tumor relapse and metastasis are primary factors contributing to breast cancer‐related deaths. Therefore, the challenge for breast cancer treatment is to sustain remission. A driving force behind tumor relapse is breast cancer heterogeneity (both intertumor, between different patients, and intratumor, within the same tumor). Understanding breast cancer heterogeneity is necessary to develop preventive interventions and targeted therapies. A recently emerging concept is that intratumor heterogeneity is driven by cancer stem cells (CSCs) that are capable of giving rise to a multitude of different cells within a tumor. Studies have highlighted linkage of CSC formation with epithelial‐to‐mesenchymal transition (EMT). In this review, we summarize the current understanding of breast cancer heterogeneity, links between EMT and CSCs, regulation of EMT by Runx transcription factors, and potential therapeutic strategies targeting these processes. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9136-9144, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26847   open full text
• Endometrial pinopode biomarkers: Molecules and microRNAs.
  Fahimeh Zamani Rarani, Fatemeh Borhani, Bahman Rashidi.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 Ultrastructural changes on the apical surface of the luminal epithelium of the uterus are known as pinopodes. Their morphology in species and in special species is associated with different results about size, duration, and percentage of surface area covered by pinopodes. The content of pinopodes is different in rodents and humans. In mice and rats pinopodes have many vacuoles and no organelle that extends to the actin stalk above the microvilli. Human pinopodes do not have a large vacuole and contain the golgi complex, a rough endoplasmic reticulum, secretory vesicles, and mitochondria that extend from the entire cell surface. It has been suggested that pinopodes are good markers of endometrial receptivity and implantation window. There are several molecular markers related to the presence of pinopodes, including integrins, leukemia inhibiting factor (LIF), l‐selectin, HOXA10, glutaredoxin, glycodelinA, heparin‐binding epidermal growth factor, mucins, and microRNAs (miRNAs). Multiple lines of evidence have indicated that miRNAs could affect the expression of LIF and pinopodes in the endometrium and these molecules play key roles in implantation window processes. Here, we have summarized the morphology and function of pinopodes. Moreover, we have highlighted several molecules in relation to pinopodes that could be used as biomarkers. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9145-9158, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26852   open full text
• High prevalence of serum IgG antibodies reacting to specific mimotopes of BK polyomavirus, a human oncogenic polyomavirus, in patients affected by uveal melanoma.
  Ilaria Bononi, Elisa Mazzoni, Silvia Pietrobon, Elena Torreggiani, Marika Rossini, Sara Violanti, Paolo Perri, Mauro Tognon, Fernanda Martini.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 The uveal melanoma (UM) is the most common human intraocular tumor. The BK polyomavirus (BKPyV) is a small DNA tumor virus whose footprints have been detected in different human cancers. BKPyV has oncogenic potential. Indeed, BKPyV, when inoculated into experimental animals, induces tumors of different histotypes, whereas in vitro, it transforms mammalian cells, including human cells from distinct tissues. In this investigation, the association between UM and BKPyV was studied employing indirect enzyme‐linked immunosorbent assays (ELISAs) using synthetic peptides that mimic BKPyV viral capsid 1 (VP1) antigens. Indirect ELISAs were used to detect serum IgG antibodies against this polyomavirus with oncogenic potential in samples from patients with UM and controls, represented by healthy subjects (HS). It was found that serum samples from patients with UM had a higher prevalence of BKPyV antibodies, 85% (51/60), compared with that detected in HS1, 62% (54/87), and HS2, 57% (68/120). The different prevalence of BKPyV antibodies detected in UM versus the two control groups, HS1 and HS2, is statistically significant (p < 0.005). Our immunologic data suggest a significantly higher prevalence of antibodies against BKPyV VP1 epitopes in serum samples from patients with UM compared with HS. These results indicate an association between UM and BKPyV, suggesting that this small DNA tumor virus may be a cofactor in the UM onset or progression. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9052-9059, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26771   open full text
• Study on the mechanism behind lncRNA MEG3 affecting clear cell renal cell carcinoma by regulating miR‐7/RASL11B signaling.
  Hongchao He, Jun Dai, Ran Zhuo, Juping Zhao, Haofei Wang, Fukang Sun, Yu Zhu, Danfeng Xu.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 The goal of this research was to study the relationships between maternally expressed gene 3 (MEG3), microRNA‐7 (miR‐7), and RASL11B, and explore their influence on the progression of clear cell renal cell carcinoma (CCRCC). Microarray analysis was conducted using the data provided by The Cancer Genome Atlas. The expression levels of MEG3 and miR‐7 in CCRCC and adjacent tissue samples were ascertained by quantitative real‐time polymerase chain reaction (qRT‐PCR). The cell proliferation activity was unmasked by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay, and cell apoptosis and cell cycle were investigated by flow cytometry. A dual luciferase reporter assay was used to verify target relationships. Wound healing assay and transwell assay were used to detect cell migration and invasion ability. Decreased MEG3 expression was observed in CCRCC tissues and cells. Overexpression of MEG3 accelerated apoptosis; inhibited cell proliferation, migration and invasion; and induced G0/G1 phase cell cycle arrest in CCRCC. MiR‐7, directly binding to MEG3, was overexpressed in the CCRCC tissues and could inhibit the apoptosis and promote the migration and invasion of CCRCC cells. RASL11B, lowly expressed in CCRCC, was a target of miR‐7. After the overexpression of RASL11B, G0/G1 phase cell cycle arrest was induced; cell apoptosis was promoted; and the proliferation, invasion, and migration of CCRCC cells were inhibited. MEG3 could up‐regulate RASL11B to inhibit the cell proliferation, invasion, and migration; induce G0/G1 cell cycle arrest; and promote cell apoptosis by suppressing miR‐7 in CCRCC. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9503-9515, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26849   open full text
• The role of hypoxia‐inducible factor‐2 alpha in angiogenesis.
  Christina Befani, Panagiotis Liakos.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2- Angiogenesis is a key enabling feature of mammalian embryonic development and tumor progression, which provides oxygen and nutrients that are required for vessel growth and tumor cell growth, respectively. Hypoxia is a driver of this phenomenon and is considered to be one of the most potent initiators of angiogenesis both in vitro and in vivo through stabilization of the transcription factors, hypoxia‐inducible factor‐1 and ‐2 (HIF‐1 and HIF‐2). Although these proteins are highly homologous, emerging evidence suggests that they have unique transcriptional targets and differential impact on angiogenesis. Although HIF‐1α is the best known and widely described isoform, recent studies suggest that HIF‐2α is a critical regulator of physiological and pathophysiological angiogenesis and, at least, the similiarly important as HIF‐1α. Indeed, HIF‐2α has been shown to regulate multiple aspects of angiogenesis, including cell proliferation, migration, maturation of blood vessels, and metastasis. In this review, we focus on recent insights into HIF‐2α expression, activation, and function under hypoxic and nonhypoxic conditions. We also summarize the current knowledge on the crosstalk between HIF‐2 and angiogenesis, describing reported phenotypical changes of HIF‐2α genetic models and HIF‐2 target genes implicated in angiogenesis. Finally, we provide a survey of recent pharmacologic strategies to specifically target HIF‐2 activity. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9087-9098, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26805   open full text
• Nucleolin protects against doxorubicin‐induced cardiotoxicity via upregulating microRNA‐21.
  Hui Sun, Zhongyi Tong, Yeqing Fang, Bimei Jiang, Pengfei Liang, Yuting Tang, Yuanbin Li, Yanyang Wu, Xianzhong Xiao.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 Nucleolin is a multifunctional protein and participates in many important biological processes. Our previous study found that nucleolin protects the heart against myocardial ischemia–reperfusion injury. In this study, we aimed to investigate the role of nucleolin in doxorubicin (DOX)‐induced cardiotoxicity. The expression pattern of nucleolin in hearts subjected to DOX injury was investigated, and we found that administration of DOX induced nucleolin expression significantly in vivo and in vitro. Gene transfection and RNA interference approaches were used in cardiomyocytes to investigate the function of nucleolin. Nucleolin overexpression protects cardiomyocytes against DOX‐induced injury. Nucleolin‐ablated cardiomyocytes become susceptible to the injury induced by DOX. The hearts of cardiac‐myocyte‐specific nucleolin transgenic mice are more resistant to DOX injury. Furthermore, nucleolin upregulates microRNA(miRNA)‐21 expression in vivo and in vitro, and the miRNA‐21 inhibitor negates the protective effect of nucleolin against injury induced by DOX. These results have demonstrated that nucleolin is involved in the regulation of DOX‐induced cardiac injury and dysfunction via the regulation of miRNA‐21 expression, and may be a novel therapeutic target for DOX‐induced cardiotoxicity. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9516-9525, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26854   open full text
• C6 ceramide motivates the anticancer sensibility induced by PKC412 in preclinical head and neck squamous cell carcinoma models.
  Yanyan Zhu, Chaojie Wang, Yun Zhou, Ning Ma, Jianwei Zhou.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2 The purpose of this study was to evaluate the anti–head and neck squamous cell carcinoma (anti‐HNSCC) cell activity by C6 ceramide and multikinase inhibitor PKC412. Experiments were performed on HNSCC cell lines (SQ20B and SCC‐9) and primary human oral carcinoma cells. Results showed that PKC412 inhibited HNSCC cell proliferation without provoking apoptosis activation. Cotreatment of C6 ceramide significantly augmented PKC412‐induced lethality in HNSCC cells. PKC412 decreased Akt–mammalian target of rapamycin (mTOR) activation in HNSCC cells, facilitated with cotreatment of C6 ceramide. In contrast, exogenous expression of a constitutively active Akt restored Akt–mTOR activation and attenuated lethality by the cotreatment. We propose that Mcl‐1 is a primary resistance factor of PKC412. The cytotoxicity of PKC412 in HNSCC cells was potentiated with Mcl‐1 short hairpin RNA knockdown, but was attenuated with Mcl‐1 overexpression. Intriguingly, C6 ceramide downregulated Mcl‐1 in HNSCC cells. In vivo, PKC412 oral administration inhibited SQ20B xenograft tumor growth in severe combined immunodeficient mice. The antitumor activity of PKC412 was further sensitized with coadministration of liposomal C6 ceramide. Together, we suggest that PKC412 could be further studied as a promising anti‐HNSCC strategy, alone or in combination with C6 ceramide. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9437-9446, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26831   open full text
• Cistanche deserticola polysaccharide attenuates osteoclastogenesis and bone resorption via inhibiting RANKL signaling and reactive oxygen species production.
  Dezhi Song, Zhen Cao, Zaibing Liu, Jennifer Tickner, Heng Qiu, Chao Wang, Kai Chen, Ziyi Wang, Shiwu Dong, Jiake Xu.
  Journal of Cellular Physiology. July 03, 2018
  --- - |2- Osteoporosis is a metabolic disease characterized by osteopenia and bone microstructural deterioration. Osteoclasts are the primary effector cells that degrade bone matrix and their abnormal function leads to the development of osteoporosis. Reactive oxygen species (ROS) accumulation during cellular metabolism promotes osteoclast proliferation and differentiation, therefore, playing an important role in osteoporosis. Cistanche deserticola polysaccharide (CDP) possesses antitumor, anti‐inflammatory, and antioxidant activity. However, the impact of CDP on osteoclasts is unclear. In this study, tartrate‐resistant acid phosphatase staining, immunofluorescence, reverse transcription‐polymerase chain reaction, and western blot analysis were utilized to demonstrate that CDP inhibited osteoclastogenesis and hydroxyapatite resorption. In addition, CDP also inhibited the expression of osteoclast maker genes including Ctsk, Mmp9, and Acp5 and had no effect on receptor activator of nuclear factor κB (RANK) expression. Mechanistic analyses revealed that CDP increases the expression of antioxidant enzymes to attenuate RANKL‐mediated ROS production in osteoclasts and inhibits nuclear factor of activated T cells and mitogen‐activated protein kinase activation. These results suggest that CDP may represent a candidate drug for the treatment of osteoporosis caused by excessive osteoclast activity. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9674-9684, December 2018.
  

  July 03, 2018   doi: 10.1002/jcp.26882   open full text
• Only a subpopulation of mouse sperm displays a rapid increase in intracellular calcium during capacitation.
  Guillermina M. Luque, Tomas Dalotto‐Moreno, David Martín‐Hidalgo, Carla Ritagliati, Lis C. Puga Molina, Ana Romarowski, Paula A. Balestrini, Liza J. Schiavi‐Ehrenhaus, Nicolas Gilio, Dario Krapf, Pablo E. Visconti, Mariano G. Buffone.
  Journal of Cellular Physiology. June 28, 2018
  --- - |2- Mammalian sperm must undergo a functionally defined process called capacitation to be able to fertilize oocytes. They become capacitated in vivo by interacting with the female reproductive tract or in vitro in a defined capacitation medium that contains bovine serum albumin, calcium (Ca2+), and bicarbonate (HCO3−). In this work, sperm were double stained with propidium iodide and the Ca2+ dye Fluo‐4 AM and analyzed by flow cytometry to determine changes in intracellular Ca2+ concentration ([Ca2+]i) in individual live sperm. An increase in [Ca2+]i was observed in a subpopulation of capacitated live sperm when compared with noncapacitated ones. Sperm exposed to the capacitating medium displayed a rapid increase in [Ca2+]i within 1 min of incubation, which remained sustained for 90 min. These rise in [Ca2+]i after 90 min of incubation in the capacitating medium was evidenced by an increase in the normalized median fluorescence intensity. This increase was dependent on the presence of extracellular Ca2+ and, at least in part, reflected the contribution of a new subpopulation of sperm with higher [Ca2+]i. In addition, it was determined that the capacitation‐associated [Ca2+]i increase was dependent of CatSper channels, as sperm derived from CatSper knockout (CatSper KO) or incubated in the presence of CatSper inhibitors failed to increase [Ca2+]i. Surprisingly, a minimum increase in [Ca2+]i was also observed in CatSper KO sperm suggesting the existence of other Ca2+ transport systems. Altogether, these results indicate that a subpopulation of sperm increases [Ca2+]i very rapidly during capacitation mainly due to a CatSper‐mediated influx of extracellular Ca2+. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9685-9700, December 2018.
  

  June 28, 2018   doi: 10.1002/jcp.26883   open full text
• RUMI is a novel negative prognostic marker and therapeutic target in non–small‐cell lung cancer.
  May Chammaa, Agnes Malysa, Carlos Redondo, Hyejeong Jang, Wei Chen, Gerold Bepler, Rodrigo Fernandez‐Valdivia.
  Journal of Cellular Physiology. June 28, 2018
  --- - |2- Recent comprehensive next‐generation genome and transcriptome analyses in lung cancer patients, several clinical observations, and compelling evidence from mouse models of lung cancer have uncovered a critical role for Notch signaling in the initiation and progression of non–small‐cell lung cancer (NSCLC). Notably, Rumi is a “protein O‐glucosyltransferase” that regulates Notch signaling through O‐glucosylation of Notch receptors, and is the only enzymatic regulator whose activity is required for both ligand‐dependent and ligand‐independent activation of Notch. We have conducted a detailed study on RUMI’s involvement in NSCLC development and progression, and have further explored the therapeutic potential of its targeting in NSCLC. We have determined that Rumi is highly expressed in the alveolar and bronchiolar epithelia, including club cells and alveolar type II cells. Remarkably, RUMI maps to the region of chromosome 3q that corresponds to the major signature of neoplastic transformation in NSCLC, and is markedly amplified and overexpressed in NSCLC tumors. Notably, RUMI expression levels are predictive of poor prognosis and survival in NSCLC patients. Our data indicates that RUMI modulates Notch activity in NSCLC cells, and that its silencing dramatically decreases cell proliferation, migration, and survival. RUMI downregulation causes severe cell cycle S‐phase arrest, increases genome instability, and induces late apoptotic–nonapoptotic cell death. Our studies demonstrate that RUMI is a novel negative prognostic factor with significant therapeutic potential in NSCLC, which embodies particular relevance especially when considering that, while current Notch inhibitory strategies target only ligand‐dependent Notch activation, a large number of NSCLCs are driven by ligand‐independent Notch activity. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9548-9562, December 2018.
  

  June 28, 2018   doi: 10.1002/jcp.26858   open full text
• Aerobic exercise, but not metformin, prevents reduction of muscular performance by AMPk activation in mice on doxorubicin chemotherapy.
  Edson A. Lima, Luís G. O. Sousa, Alexandre Abilio Teixeira, Andrea G. Marshall, Nelo E. Zanchi, José C. Rosa Neto.
  Journal of Cellular Physiology. June 28, 2018
  --- - |2- Doxorubicin (DOX) is a chemotherapy agent widely used in clinical practice, and it is very efficient in tumor suppression, but the use of DOX is limited by a strong association with the development of severe muscle atrophy and cardiotoxicity effects. Reversion or neutralization of the muscular atrophy can lead to a better prognosis. Recent studies have proposed that the negative effect of DOX on skeletal muscle is linked to its inhibition of AMP‐activated protein kinase (AMPk), a key mediator of cellular metabolism. On the basis of this, our goal was to evaluate if aerobic exercise or metformin treatment, activators of AMPk, would be able to attenuate the deleterious effects on skeletal muscle induced by the DOX treatment. C57BL6 mice received either saline (control) or DOX (2.5 mg/kg body weight) intraperitoneally, twice a week. The animals on DOX were further divided into groups that received adjuvant treatment in the form of moderate aerobic physical exercise (DOX+T) or metformin gavage (300 mg/body weight/day). Body weight, metabolism, distance run, muscle fiber cross‐sectional area (CSA), and protein synthesis and degradation were assessed. We demonstrated that aerobic training, but not metformin, associated with DOX increased the maximal aerobic capacity without changing muscle mass or fiber CSA, rescuing the muscle fatigue observed with DOX treatment alone. This improvement was associated with AMPk activation, thus surpassing the negative effects of DOX on muscle performance and bioenergetics. In conclusion, aerobic exercise increases AMPk activation and improved the skeletal muscle function, reducing the side effects of DOX. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9652-9662, December 2018.
  

  June 28, 2018   doi: 10.1002/jcp.26880   open full text
• HOXA11‐AS promotes the growth and invasion of renal cancer by sponging miR‐146b‐5p to upregulate MMP16 expression.
  Feng‐Qiang Yang, Jian‐Qiu Zhang, Jiang‐Jiang Jin, Chong‐Yi Yang, Wei‐Jie Zhang, Hai‐Ming Zhang, Jun‐Hua Zheng, Ze‐Ming Weng.
  Journal of Cellular Physiology. June 28, 2018
  --- - |2- Recently, increasing studies showed that long noncoding RNAs (lncRNAs) play critical roles in tumor progression. However, the function and underlying mechanism of HOMEOBOX A11 antisense RNA (HOXA11‐AS) on renal cancer remain unclear. In the current study, our data showed that the expression of HOXA11‐AS was significantly upregulated in clear cell renal cell carcinoma (ccRCC) tissues and cell lines. High HOXA11‐AS expression was associated with the advanced clinical stage, tumor stage, and lymph node metastasis. Function assays showed that HOXA11‐AS inhibition significantly suppressed renal cancer cells growth, invasion, and ETM phenotype. In addition, underlying mechanism revealed that HOXA11‐AS could act as a competing endogenous RNA (ceRNA) that repressed miR‐146b‐5p expression, which regulated its downstream target MMP16 in renal cancer. Taken together, our findings suggested that HOXA11‐AS could promote renal cancer cells growth and invasion by modulating miR‐146b‐5p–MMP16 axis. Thus, our findings suggested that HOXA11‐AS could serve as potential therapeutic target for the treatment of renal cancer. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9611-9619, December 2018.
  

  June 28, 2018   doi: 10.1002/jcp.26864   open full text
• NLRP3 inflammasome mediates chronic intermittent hypoxia‐induced renal injury implication of the microRNA‐155/FOXO3a signaling pathway.
  Xu Wu, Su Chi Chang, Jifu Jin, Wenyu Gu, Shanqun Li.
  Journal of Cellular Physiology. June 28, 2018
  --- - |2- Chronic intermittent hypoxia (CIH), as the foremost pathophysiological change of obstructive sleep apnea (OSA), contributes to continued deterioration in renal function. Nucleotide‐binding domain like receptor protein 3 (NLRP3) inflammasome is a multiprotein complex that triggers innate immune responses to infection and cell stress through activation of caspase‐1 and maturation of inflammatory pro‐interleukin‐1β cytokine. Emerging evidence indicates that inhibition of the NLRP3 inflammasome ameliorates renal injury. Nevertheless, it is uncertain whether NLRP3 inflammasome participates in CIH‐induced renal injury. The molecular mechanisms modulating NLRP3 inflammasome activation remain to be elucidated. Compared with wild‐type mice, NLRP3 knockout mice dramatically protected them from kidney injury, as indicated by the restoration of creatinine levels, lessened histopathological alterations, and the suppression of macrophages infiltration stained with F4/80. NLRP3 deficiency notably reversed CIH‐induced oxidative stress (malondialdehyde and superoxide dismutase), concomitantly with the abrogated apoptosis‐related proteins and proinflammatory signaling pathway. Consistently, NLRP3‐deficient tubular cells remarkably inhibited reactive oxygen species generation and NLRP3 inflammasome activation. Furthermore, our study revealed that microRNA‐155 (miR‐155) was augmented in the renal tissue and HK‐2 cells exposed to CIH. In addition, we investigated the role of miR‐155 in the regulation of NLRP3 inflammasome. Inhibition of miR‐155 suppressed the CIH‐induced NLRP3 inflammasome activation in renal tubular cells, whereas overexpression of miR‐155 promoted oxidation and enhanced NLRP3 pathway. Collectively, we demonstrated that miR‐155 might be a positive‐regulator of NLRP3 pathway by inhibiting the targeted FOXO3a gene. These results established a link between the miR‐155/FOXO3a pathway and the NLRP3 inflammasome, suggesting pharmacological blockage of NLRP3 as a potential therapeutic strategy for OSA‐associated chronic kidney disease. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9404-9415, December 2018.
  

  June 28, 2018   doi: 10.1002/jcp.26784   open full text
• Fibroblasts and mesenchymal stem cells: Two sides of the same coin?
  Mathangi Soundararajan, Suresh Kannan.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2 Mesenchymal stem/stromal cells (MSCs) have gained considerable popularity owing to the vast possibilities and lack of ethical constraints and risks normally associated with other stem cells, such as embryonic stem cells. However, they are morphologically indistinguishable from fibroblasts. This review aims to assess the similarities and differences between the two cell types, and the possible relationship between them. We found that the two cells seem almost identical with respect to their surface immunophenotype, proliferation, and differentiation capacities and even, to an extent, their gene expression profiles and immunomodulatory capacities. There are some differences in capability between the two cells, with MSCs being more efficient than fibroblasts. Even so, the similarities are so striking, that, if we were to follow the current criteria provided by the International Society for Cellular Therapy, fibroblasts ought to be named as MSCs. One promising marker is their DNA methylation profiles. Nonetheless, without any other marker to differentiate between the cells in the first place, it would be difficult to find a definitive marker. Interestingly, the differences observed between the two cells have also been observed between young and old MSCs. This also seems to be true of certain cell surface markers. Therefore, it is possible that fibroblasts are in fact aged MSCs and that the two cells are the same. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9099-9109, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26860   open full text
• Adipokines, adiposity, and bone marrow adipocytes: Dangerous accomplices in multiple myeloma.
  Emma V. Morris, Claire M. Edwards.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- Obesity has become a global epidemic influencing the establishment and progression of a wide range of diseases, such as diabetes, cardiovascular disease, and cancer. In 2016, International Agency for Research on Cancer reported that obesity is now associated with 13 different cancers, one of which is multiple myeloma (MM), a destructive cancer of plasma cells that predominantly reside in the bone marrow. Obesity is the accumulation of excess body fat, which causes metabolic, endocrine, immunologic, and inflammatory‐like changes. Obesity is usually associated with an increase in visceral and/or subcutaneous fat; however, an additional fat depot that also responds to diet‐induced changes is bone marrow adipose tissue (BMAT). There have been several studies over the past few decades that have identified BMAT as a key driver in MM progression. Adipocytes secrete numerous adipokines, such as leptin, adiponectin, resistin, adipsin, and visfatin, which when secreted at normal controlled levels have protective properties. However, in obesity these levels of secretion change, coupled with an increase in adipocyte number and size causing a profound and lasting effect on the bone microenvironment, contributing to MM cell growth, survival, and migration as well as potentially fueling bone destruction. Obesity is a modifiable risk factor making it an attractive option for targeted therapy. This review discusses the link between obesity, monoclonal gammopathy of undetermined significance (a benign condition that precedes MM), and myeloma, and the contribution of key adipokines to disease establishment and progression. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9159-9166, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26884   open full text
• Enhancing developmental rate and quality of mouse single blastomeres into blastocysts using a microplatform.
  Farshid Yekani, Mohammad Fazel‐Tabar, Reza Kowsari‐Esfahan, Philippe Renaud, Hanieh Kavand, Fereshteh Esfandiari, Mahnaz Azarnia, Leila Montazeri, Hossein Baharvand.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- The present work reports the beneficial effects of using a microplatform on the development of mouse single blastomeres (SBs) to the blastocyst stage. Development of blastocysts from SBs separated from two‐ and four‐cell stage embryos (two‐ and four‐cell SBs) can provide a valuable supply both for couples who use fertility‐assisted techniques and farm animals. As a step forward, we introduce three chips that provide the possibility of culturing SBs separately, in groups, and in the vicinity of the intact embryo (co‐culture), while each well of the chips is assigned to an isolated SB. Two‐ and four‐cell SBs co‐cultured with intact embryos showed 97.1% and 76.6% developmental rates and up to 34.1% and 49.1% growth relative to the microdroplet method (control). We examined the quality of developed blastocysts by assessing the total cell number, the number of inner cell mass (ICM) according to the octamer‐binding transcription factor 4 marker (OCT4), and trophectoderm (TE). Co‐culture of SBs with an intact embryo in a chip with nanoscale culture medium volume also increased the cell population of the developed embryo. The ICM:TE ratio, which is the most important blastocyst quality parameter, also indicated that developed two‐cell SBs have a higher degree of similarity to intact embryos despite fewer numbers of total cells. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9070-9076, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26879   open full text
• Role of IL‐6‐mediated expression of NS5ATP9 in autophagy of liver cancer cells.
  Hongping Lu, Ming Han, Xiaoxue Yuan, Kelbinur Tursun, Yu Zhang, Yaru Li, Zhongshu Li, Shenghu Feng, Li Zhou, Zhipeng Pan, Qi Wang, Kai Han, Shunai Liu, Jun Cheng.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2+ This study aimed to investigate the relationship between interleukin‐6 (IL‐6) and NS5ATP9 in autophagy of liver cancer cells. Autophagy is one of the important regulators of the replication of hepatitis C virus and the survival of tumors. IL‐6 is a multifunctional cytokine that plays an important role in autophagy and development of many kinds of tumors. However, the role of IL‐6 in autophagy has not been fully explored. A previous study had shown that a novel gene, NS5ATP9, could modulate autophagy. The present study demonstrated that human IL‐6 recombinant protein induced autophagy of HepG2 cells. Conversely, autophagy decreased after IL‐6 was silenced or neutralized with monoclonal antibody against human IL‐6. In addition, NS5ATP9 was upregulated by IL‐6 via nuclear factor–kappaB activation, as detected by Western blot. Further studies indicated that the induction of autophagy by IL‐6 could be attenuated by silencing NS5ATP9. Interestingly, the expression of NS5ATP9, in turn, resulted in the upregulation of IL‐6. In conclusion, IL‐6 could induce autophagy by expressing NS5ATP9, while NS5ATP9 upregulated IL‐6 levels in turn, which further induced autophagy. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9312-9319, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26343   open full text
• PMLIV overexpression promotes TGF‐β‐associated epithelial–mesenchymal transition and migration in MCF‐7 cancer cells.
  Yu Liu, Jia‐Xin Wang, Di Huang, Bing Wang, Liang‐Liang Li, Xiu‐Xian Li, Ping Ni, Xing‐Li Dong, Wei Xia, Chun‐Xiao Yu, Wan‐Lu Xu, Wen‐Feng Chu, Dan Zhao.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- The epithelial–mesenchymal transition (EMT) is a key event associated with metastasis and dissemination in breast tumor pathogenesis. Promyelocytic leukemia (PML) gene produces several isoforms due to alternative splicing; however, the biological function of each specific isoform has yet to be identified. In this study, we report a previously unknown role for PMLIV, the most intensely studied nuclear isoform, in transforming growth factor‐β (TGF‐β) signaling‐associated EMT and migration in breast cancer. This study demonstrates that PMLIV overexpression promotes a more aggressive mesenchymal phenotype and increases the migration of MCF‐7 cancer cells. This event is associated with activation of the TGF‐β canonical signaling pathway through the induction of Smad2/3 phosphorylation and the translocation of phospho‐Smad2/3 to the nucleus. In this study, we report a previously unknown role for PMLIV in TGF‐β signaling‐induced regulation of breast cancer‐associated EMT and migration. Targeting this pathway may be therapeutically beneficial. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9575-9583, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26862   open full text
• Cleavage of caspase‐12 at Asp94, mediated by endoplasmic reticulum stress (ERS), contributes to stretch‐induced apoptosis of myoblasts.
  Jing Song, Qiang Zhang, Shuai Wang, Fang Yang, Zhenggang Chen, Quanjiang Dong, Qiuxia Ji, Xiao Yuan, Dapeng Ren.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- Mechanical overloading can lead to skeletal muscle damage instead of remodeling. This is attributed to the excessive apoptosis of myoblasts, mechanism of which remains to be elucidated. The present study aimed to investigate the involvement of endoplasmic reticulum stress (ERS) and caspase‐12 in mediating the stretch‐induced apoptosis of myoblasts. Myoblast apoptosis was evaluated by Hoechst staining, DNA fragmentation assay, Annexin V binding, and propidium iodide staining, as well as caspase‐3 and poly‐ADP‐ribose polymerase 1 cleavage. First, our results showed that apoptosis was elevated in a time‐dependent manner when myoblasts were subjected to cyclic mechanical stretch (CMS) for 12, 24, and 36 hr. Concomitantly, CMS triggered the ERS and caspase‐12 cleavage; ERS inhibitor GSK 2606414 suppressed the CMS‐induced cleavage of caspase‐12 and myoblast apoptosis. Silencing caspase‐12 attenuated the apoptosis of myoblasts under CMS. Furthermore, CMS‐induced myoblast apoptosis was partially recovered by overexpressing wild‐type caspase‐12 in caspase‐12‐silenced myoblasts. In contrast, overexpressing mutant caspase‐12 (D94N), which cannot be cleaved into the active caspase‐12 fragments, failed to accomplish the same effect. Finally, C2C12 overexpressing truncated caspase‐12 segment (TC‐casp12‐D94), which starts from Asp94 and ends at Asn419, underwent apoptosis under both static and stretched conditions. Interestingly, C2C12 myoblasts seemed to be resistant to stretch‐induced apoptosis upon low‐serum‐induced differentiation. In conclusion, our study provided evidence that caspase‐12 cleavage at Asp94, induced by ERS under mechanical stimuli, is the key molecule in initiating the stretch‐triggered apoptosis of myoblasts. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9473-9487, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26840   open full text
• ΔFosB regulates rosiglitazone‐induced milk fat synthesis and cell survival.
  Xuefeng Wei, Hui Li, Guangwei Zhao, Jiameng Yang, Lihui Li, Yongzhen Huang, Xianyong Lan, Yun Ma, Linyong Hu, Huiling Zheng, Hong Chen.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2+ Rosiglitazone induces adipogenesis in adipocyte and regulates cell survival and differentiation in number of cell types. However, whether PPARγ regulates the synthesis of milk fat and cell survival in goat mammary gland remains unknown. Rosiglitazone strongly enhanced cellular triacylglycerol content and accumulation of lipid droplet in goat mammary epithelial cells (GMEC). Furthermore, ΔFosB decreased the expression of PPARγ at both mRNA and protein levels, and rosiglitazone‐induced milk fat synthesis was abolished by ΔFosB overexpression. ΔFosB reduced milk fat synthesis and enhanced saturated fatty acid concentration. Rosiglitazone increased the number of GMEC in G0/G1 phase and inhibited cell proliferation, and these effects were improved by overexpression of ΔFosB. ΔFosB was found to promote the expression of Bcl‐2 and suppress the expression of Bax, and protected GMEC from apoptosis induced by rosiglitazone. Intracellular calcium trafficking assay revealed that rosiglitazone markedly increased intracellular calcium concentration. ΔFosB protected GMEC from apoptosis induced by intracellular Ca2+ overload. ΔFosB increased MMP‐9 gelatinolytic activity. SB‐3CT, an MMP‐9 inhibitor, suppressed the expression of Bcl‐2, and increased intracellular calcium levels, and this effect was abolished by ΔFosB overexpression. SB‐3CT induced GMEC apoptosis and this effect was inhibited by ΔFosB overexpression. These findings suggest that ΔFosB regulates rosiglitazone‐induced milk fat synthesis and cell survival. Therefore, ΔFosB may be an important checkpoint to control milk fat synthesis and cell apoptosis. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9284-9298, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26218   open full text
• Oxidative stress regulates autophagy in cultured muscle cells of patients with chronic obstructive pulmonary disease.
  Fares Gouzi, Marine Blaquière, Matthias Catteau, François Bughin, Jonathan Maury, Emilie Passerieux, Bronia Ayoub, Jacques Mercier, Maurice Hayot, Pascal Pomiès.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- The proteolytic autophagy pathway is enhanced in the lower limb muscles of patients with chronic obstructive pulmonary disease (COPD). Reactive oxygen species (ROS) have been shown to regulate autophagy in the skeletal muscles, but the role of oxidative stress in the muscle autophagy of patients with COPD is unknown. We used cultured myoblasts and myotubes from the quadriceps of eight healthy subjects and twelve patients with COPD (FEV1% predicted: 102.0% and 32.0%, respectively; p < 0.0001). We compared the autophagosome formation, the expression of autophagy markers, and the autophagic flux in healthy subjects and the patients with COPD, and we evaluated the effects of the 3‐methyladenine (3‐MA) autophagy inhibitor on the atrophy of COPD myotubes. Autophagy was also assessed in COPD myotubes treated with an antioxidant molecule, ascorbic acid. Autophagosome formation was increased in COPD myoblasts and myotubes (p = 0.011; p < 0.001), and the LC3 2/LC3 1 ratio (p = 0.002), SQSTM1 mRNA and protein expression (p = 0.023; p = 0.007), BNIP3 expression (p = 0.031), and autophagic flux (p = 0.002) were higher in COPD myoblasts. Inhibition of autophagy with 3‐MA increased the COPD myotube diameter (p < 0.001) to a level similar to the diameter of healthy subject myotubes. Treatment of COPD myotubes with ascorbic acid decreased ROS concentration (p < 0.001), ROS‐induced protein carbonylation (p = 0.019), the LC3 2/LC3 1 ratio (p = 0.037), the expression of SQSTM1 (p < 0.001) and BNIP3 (p < 0.001), and increased the COPD myotube diameter (p < 0.001). Thus, autophagy signaling is enhanced in cultured COPD muscle cells. Furthermore, the oxidative stress level contributes to the regulation of autophagy, which is involved in the atrophy of COPD myotubes in vitro. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9629-9639, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26868   open full text
• P2Y12 shRNA treatment decreases SGC activation to relieve diabetic neuropathic pain in type 2 diabetes mellitus rats.
  Shouyu Wang, Zilin Wang, Lin Li, Lifang Zou, Yingxin Gong, Tianyu Jia, Shanhong Zhao, Huilong Yuan, Liran Shi, Shuangmei Liu, Bing Wu, Zhihua Yi, Hui Liu, Yun Gao, Guilin Li, Jan M. Deussing, Man Li, Chunping Zhang, Shangdong Liang.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- Diabetic neuropathic pain is a common complication of type 2 diabetes mellitus (DM). Activation of satellite glial cells (SGCs) in the dorsal root ganglia (DRG) plays a crucial role in neuropathic pain through the release of proinflammatory cytokines. The P2Y12 receptor is expressed in SGCs of the DRG. In this study, our aim was to investigate the role of the P2Y12 receptor on the pathological changes in diabetic neuropathic pain. The present study showed that diabetic neuropathic pain increased mechanical and thermal hyperalgesia in type 2 DM model rats. The results showed that the expression levels of P2Y12 messenger RNA (mRNA) and protein in DRG SGCs were increased in DM model rats compared with control rats. Glial fibrillary acidic protein (GFAP) and interleukin‐1β (IL‐1β) expression levels in the DRG were increased in DM rats. Upregulation of GFAP is a marker of SGC activation. Targeting the P2Y12 receptor by short hairpin RNA (shRNA) decreased the upregulated expression of P2Y12 mRNA and protein, coexpression of P2Y12 and GFAP, the expression of GFAP, IL‐1β, and tumor necrosis factor‐receptor 1 in the DRG of DM rats, and relieved mechanical and thermal hyperalgesia in DM rats. After treatment with the P2Y12 receptor shRNA, the enhancing integrated OPTICAL density (IOD) ratios of p‐P38 MAPK to P38 mitogen activated protein kinase (MAPK) in the DM rats treated with P2Y12 shRNA were significantly lower than that in the untreated DM rats. Therefore, P2Y12 shRNA treatment decreased SGC activation to relieve mechanical and thermal hyperalgesia in DM rats. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9620-9628, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26867   open full text
• A new copper ionophore DPMQ protects cells against ultraviolet B irradiation by inhibiting the TRPV1 channel.
  Kuo‐Feng Huang, Kuo‐Hsing Ma, Yu‐Chien Hung, Liang‐Chuan Lo, Kuo‐Chen Lin, Pei‐Shan Liu, Ming‐Kuan Hu, Sheau‐Huei Chueh.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- Copper is more likely than iron to generate reactive oxygen species (ROS) in a redox reaction due to its higher electrochemical reactivity. This study examined the effect of a newly synthesized Cu2+ binding compound, (E)‐2‐(4‐(dimethylamino)phenylimino)methyl)quinolin‐8‐ol (DPMQ), on ultraviolet B (UVB) irradiation–induced cytotoxicity in human dermal fibroblasts. DPMQ induced Cu2+ influx as effectively as disulfiram, a Cu2+ ionophore anticancer drug. However, disulfiram induced ROS generation, mitochondrial dysfunction, and apoptosis in fibroblasts in a Cu2+‐dependent manner, whereas DPMQ was not only nontoxic, but protected cells against UVB irradiation–induced apoptosis in a Cu2+‐independent manner. UVB irradiation induced a Ca2+‐dependent increase in ROS generation, a decrease in Nrf2 levels, and activation of the mitochondrial apoptotic pathway, and these effects were prevented by DPMQ, which also increased Nrf2 nuclear translocation in a Cu2+‐independent manner. UVB irradiation activated 12‐lipoxygenase and 12‐hydroxyeicosatetraenoic acid (12‐HETE), a product of 12‐lipoxygenase, activated the TRPV1 channel. DMPQ did not act as a Ca2+ chelator, but inhibited the cytosolic Ca2+ increase induced by 12‐HETE or capsaicin, but not that induced by bradykinin or ATP. Blockade of Ca2+ influx by pharmacological inhibition or silencing of the TRPV1 channel or chelation of cytosolic Ca2+ inhibited the UVB irradiation–induced Nrf2 reduction, ROS generation, mitochondrial dysfunction, and apoptosis. Taken together, our results suggest that Ca2+ influx via the TRPV1 channel is responsible for UVB irradiation–induced cytotoxicity and that DPMQ protects cells against UVB irradiation by inhibiting the TRPV1 channel and stabilizing Nrf2, and could thus be a potentially useful compound for the treatment of free radical‐induced diseases. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9594-9610, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26861   open full text
• Zerumbone inhibits epithelial‐mesenchymal transition and cancer stem cells properties by inhibiting the β‐catenin pathway through miR‐200c.
  Fatemeh Karimi Dermani, Razieh Amini, Massoud Saidijam, Mona Pourjafar, Sahar Saki, Rezvan Najafi.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- Colorectal cancer (CRC) is one of the most lethal and rampant human malignancies in the world. Zerumbone, a sesquiterpene isolated from subtropical ginger, has been found to exhibit an antitumor effect in various cancer types. However, the effect of Zerumbone on the biological properties of CRC, including epithelial‐mesenchymal transition (EMT) and cancer stem cells (CSCs) has not been fully elucidated. Here, we investigated the inhibitory action of Zerumbone on the EMT process, CSC markers, and the β‐catenin signaling pathway in the presence or absence of miR‐200c. The effect of Zerumbone on HCT‐116 and SW‐48 cells viability was examined by 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyltetrazolium bromide assay. The effects of Zerumbone on EMT‐related genes, CSCs markers, cell migration, invasion, sphere‐forming, and β‐catenin signaling pathway were explored. To evaluate the role of miR‐200c in anticancer effects by Zerumbone, miR‐200c was downregulated by LNA‐anti‐miR‐200c. Zerumbone significantly inhibited cell viability, migration, invasion, and sphere‐forming potential in HCT‐116 and SW‐48 cell lines. Zerumbone significantly suppressed the EMT and CSC properties as well as downregulated the β‐catenin. Silencing of miR200c reduced the inhibitory effects of Zerumbone on EMT and CSCs in CRC cells. These data indicated that Zerumbone may be a promising candidate for reducing the risk of CRC progression by suppressing the β‐catenin pathway via miR‐200c. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9538-9547, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26874   open full text
• Influence of all‐trans retinoic acid on sperm metabolism and oxidative stress: Its involvement in the physiopathology of varicocele‐associated male infertility.
  Rocco Malivindi, Vittoria Rago, Daniela De Rose, Maria Clelia Gervasi, Erika Cione, Giampiero Russo, Marta Santoro, Saveria Aquila.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2 The mechanisms by which varicocele affects fertility remain undetermined. Vitamin A (all‐trans retinoic acid [ATRA]) is required for fertility and normal spermatogenesis; however, the mechanisms driving its action are not defined yet. Previously, we demonstrated in varicocele sperm a reduced RARα expression and that ATRA influence sperm performance. To further define vitamin A significance in male gamete and in the physiopathology of varicocele, we tested for the first time ATRA action on human sperm metabolism and antioxidant defense systems. Evaluating triglycerides content and lipase activity, in normal sperm ATRA had a lipid lowering effect, which was not observed in varicocele sperm. The modulation of the glucose‐6‐phosphate dehydrogenase activity, concomitantly with a reduction of the glucose content, highlight an ATRA role on glucose metabolism. ATRA induced the superoxide dismutase (SOD) and glutathione transferase activities, while it reduced the malondialdehyde and reactive oxygen species (ROS) production both in healthy and varicocele sperm. Interestingly, SOD1 and SOD2 have been localized in the acrosome and midpiece, glutathione‐ S‐transferase omega 2 (GSTO2) in the acrosome, equatorial, and subacrosomial regions. SOD1, SOD2, and GSTO2 levels were significantly lower in varicocele with respect to healthy sperm. Herein, we discovered that ATRA treatment was able to reprogram sperm metabolism toward that of the capacitation status. The retinol protected human sperm from ROS damage enhancing the antioxidant enzymes activity, providing evidence toward the efficacy of vitamin A as therapeutic tool in improving sperm quality. These novel findings further confirm the importance of vitamin A in male fertility adding new insights into the retinoids complex biological framework. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9526-9537, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26872   open full text
• BMAL1 and CLOCK proteins in regulating UVB‐induced apoptosis and DNA damage responses in human keratinocytes.
  Yang Sun, Peiling Wang, Hongyu Li, Jun Dai.
  Journal of Cellular Physiology. June 26, 2018
  --- - |2- A diverse array of biological processes are under circadian controls. In mouse skin, ultraviolet ray (UVR)‐induced apoptosis and DNA damage responses are time‐of‐day dependent, which are controlled by core clock proteins. This study investigates the roles of clock proteins in regulating UVB responses in human keratinocytes (HKCs). We found that the messenger RNA expression of brain and muscle ARNT‐like 1 (BMAL1) and circadian locomotor output cycles kaput (CLOCK) genes is altered by low doses (5 mJ/cm2) of UVB in the immortalized HaCat HKCs cell line. Although depletion of BMAL1 or CLOCK has no effect on the activation of Rad3‐related protein kinases–checkpoint kinase 1–p53 mediated DNA damage checkpoints, it leads to suppression of UVB‐stimulated apoptotic responses, and downregulation of UVB‐elevated expression of DNA damage marker γ‐H2AX and cell cycle inhibitor p21. Diminished apoptotic responses are also observed in primary HKCs depleted of BMAL1 or CLOCK after UVB irradiation. While CLOCK depletion shows a suppressive effect on UVB‐induced p53 protein accumulation, depletion of either clock gene triggers early keratinocyte differentiation of HKCs at their steady state. These results suggest that UVB‐induced apoptosis and DNA damage responses are controlled by clock proteins, but via different mechanisms in the immortalized human adult low calcium temperature and primary HKCs. Given the implication of UVB in photoaging and photocarcinogenesis, mechanistic elucidation of circadian controls on UVB effects in human skin will be critical and beneficial for prevention and treatment of skin cancers and other skin‐related diseases. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9563-9574, December 2018.
  

  June 26, 2018   doi: 10.1002/jcp.26859   open full text
• Gut microbiota‐derived endotoxin enhanced the incidence of cardia bifida during cardiogenesis.
  Jing Zhang, Guang Wang, Jia Liu, Lin‐rui Gao, Meng Liu, Chao‐jie Wang, Manli Chuai, Yongping Bao, Ge Li, Rui‐man Li, Yu Zhang, Xuesong Yang.
  Journal of Cellular Physiology. June 22, 2018
  --- - |2+ Cytotoxicity and inflammation‐associated toxic responses could be induced by bacterial lipopolysaccharides (LPS) in vitro and in vivo, respectively. However, the mechanism involved in LPS‐induced cardiac malformation in prenatal fetus is still unknown. In this study, we demonstrated that LPS was induced in gut microbiota imbalance mice, and next, LPS exposure during gastrulation in the chick embryo increased the incidence of cardia bifida. Gene transfection and tissue transplantation trajectory indicated that LPS exposure restricted the cell migration of cardiac progenitors to primary heart field in gastrula chick embryos. In vitro explant allograft of GFP‐labeled anterior primitive streak demonstrated that LPS treatments could inhibit cell migration. A similar observation was also obtained from the cell migration assay of scratch wounds using primary culture of cardiomyocytes or H9c2 cells. In the embryos exposed to LPS, expressions of Nkx2.5 and GATA5 were disturbed. These genes are associated with cardiomyocyte differentiation when heart tube fusion occurs. Furthermore, pHIS3, C‐caspase3 immunohistological staining indicated that cell proliferation decreased, cell apoptosis increased in the heart tube of chick embryo. Meanwhile, in vivo, pHIS3 immunohistological staining and Hochest/PI staining also draw the similar conclusions. The LPS exposure also caused the production of excess ROS, which might damage the cardiac precursor cells of developing embryos. At last, we showed that LPS‐induced cardia bifida could be partially rescued through the addition of antioxidants. Together, these results reveal that excess ROS generation is involved in the LPS‐induced defects in heart tube during chick embryo development. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9271-9283, December 2018.
  

  June 22, 2018   doi: 10.1002/jcp.26175   open full text
• Serum C‐reactive protein in the prediction of cardiovascular diseases: Overview of the latest clinical studies and public health practice.
  Amir Avan, Seyedeh Belin Tavakoly Sany, Majid Ghayour‐Mobarhan, Hamid Reza Rahimi, Mohammad Tajfard, Gordon Ferns.
  Journal of Cellular Physiology. June 22, 2018
  --- - |2- Cardiovascular disease is the most common cause of morbidity and mortality globally. Epidemiological studies using high‐sensitivity assays for serum C‐reactive protein have shown a consistent association between cardiovascular disease risk and serum C‐reactive protein concentrations. C‐reactive protein is a biomarker for inflammation, and has been established in clinical practice as an independent risk factor for cardiovascular disease events. There is evidence that serum C‐reactive protein is an excellent biomarker of cardiovascular disease and is also an independent and strong predictor of adverse cardiovascular events. Further characterization of the impact and influence of lifestyle exposures and genetic variation on the C‐reactive protein response to cardiovascular disease events may have implications for the therapeutic approaches to reduce cardiovascular disease events. This review summarizes the studies that have examined the association between serum C‐reactive protein and the risk of cardiovascular disease. We also discuss the impact of independent factors and C‐reactive protein genetic polymorphisms on baseline plasma C‐reactive protein levels. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8508-8525, November 2018.
  

  June 22, 2018   doi: 10.1002/jcp.26791   open full text
• Potential role of microRNAs in the regulation of adipocytes liposecretion and adipose tissue physiology.
  Giulia Maurizi, Lucia Babini, Lucio Della Guardia.
  Journal of Cellular Physiology. June 22, 2018
  --- - |2+ Adipose tissue is a dynamic endocrine organ playing a pivotal role in metabolism modulation. Adipocytes differentiation requires a highly orchestrated series of changes of gene expression in precursor cells. At the same time, white mature adipocytes are plastic cells able to reversibly transdifferentiate toward fibroblast‐like cells via the liposecretion process, returning back to a non‐committed status of the cells. In particular, adipose tissue microenvironment along with external signaling molecules such as adipokines, cytokines and growth factors can regulate adipocytes physiology through complex molecular networks. MicroRNAs (miRNAs), a type of non‐coding RNA, acting as fine regulators of biological processes and their expression is sensible to the environment and cellular status changes. MiRNAs are thought to play a pivotal role in regulating the physiology of adipose tissue as well as in the development of obesity and associated metabolic disturbances, although the underlying mechanisms have not been identified so far. Elucidating the molecular mechanisms orchestrating adipose tissue biology is required to better characterize obesity and its associated diseases. In this respect, the review aims to analyze the microRNAs potentially involved in adipogenesis highlighting their role in the process of liposecretion, adipocyte proliferation, and adipokines secretion. The role of microRNAs in the development of obesity and obesity‐associated disorders is also discussed. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9077-9086, December 2018.
  

  June 22, 2018   doi: 10.1002/jcp.26523   open full text
• Network analysis of hippocampal neurons by microelectrode array in the presence of HIV‐1 Tat and cocaine.
  Taha Mohseni Ahooyi, Masoud Shekarabi, Emilie A. Decoppet, Dianne Langford, Kamel Khalili, Jennifer Gordon.
  Journal of Cellular Physiology. June 22, 2018
  --- - |2+ HIV‐associated neurocognitive disorders affecting greater than 30% of patients are caused by HIV‐1 infection of the CNS, and in part, include neurotoxic effects of the viral transactivator of transcription, Tat protein. In addition to increasing the risk for becoming HIV infected, cocaine abuse enhances the neuropathogenic impacts of HIV‐1. To investigate the outcome of Tat and cocaine interference in the hippocampal neuronal network, cross‐rank‐corrlation was employed to develop a systematic framework to assess hippocampal neurons behavior cultured on multielectrode arrays. Tat and cocaine differentially disturbed neuronal spiking rates, amplitude, synchronous activity, and oscillations within the hippocampal neuronal network via potentiation of inhibitory neurotransmission. The Tat‐mediated impairment of neuronal spiking was reversible by removal of Tat, which restored neuronal activity. The presence of astrocytes co‐cultured with neuronal networks diminished the effects of Tat and cocaine on neuron function suggesting a role for astrocytes in stabilizing neuronal behavior and increasing neuronal spontaneous activities such as bursting amplitude, frequency, and wave propagation rate. Taken together, our studies indicate that the HIV protein Tat and cocaine impair hippocampal neuronal network functioning and that the presence of astrocytes alleviates network dysfunction pointing to a newly discovered pathway through which ionic homeostasis is maintained by neuron‐glial crosstalk in the CNS. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9299-9311, December 2018.
  

  June 22, 2018   doi: 10.1002/jcp.26322   open full text
• EGFL7: Master regulator of cancer pathogenesis, angiogenesis and an emerging mediator of bone homeostasis.
  Guoju Hong, Vincent Kuek, Jiaxi Shi, Lin Zhou, Xiaorui Han, Wei He, Jennifer Tickner, Heng Qiu, Qiushi Wei, Jiake Xu.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2 Epidermal growth factor‐like domain‐containing protein 7 (EGFL7), a member of the epidermal growth factor (EGF)‐like protein family, is a potent angiogenic factor expressed in many different cell types. EGFL7 plays a vital role in controlling vascular angiogenesis during embryogenesis, organogenesis, and maintaining skeletal homeostasis. It regulates cellular functions by mediating the main signaling pathways (Notch, integrin) and EGF receptor cascades. Accumulating evidence suggests that Egfl7 plays a crucial role in cancer biology by modulating tumor angiogenesis, metastasis, and invasion. Dysregulation of Egfl7 has been frequently found in several types of cancers, such as malignant glioma, colorectal carcinoma, oral and oesophageal cancers, gastric cancer, hepatocellular carcinoma, pancreatic cancer, breast cancer, lung cancer, osteosarcoma, and acute myeloid leukemia. In addition, altered expression of miR‐126, a microRNA associated with Egfl7, was found to play an important role in oncogenesis. More recently, our study has shown that EGFL7 is expressed in both the osteoclast and osteoblast lineages and promotes endothelial cell activities via extracellular signal‐regulated kinase (ERK), signal transducer and activator of transcription 3 (STAT3), and integrin signaling cascades, indicative of its angiogenic regulation in the bone microenvironment. Thus, understanding the role of EGFL7 may provide novel insights into the development of improved diagnostics and therapeutic treatment for cancers and skeletal pathological disorders, such as ischemic osteonecrosis and bone fracture healing. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8526-8537, November 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26792   open full text
• Circulating microRNAs as diagnostic and therapeutic biomarkers in gastric and esophageal cancers.
  Leila Jamali, Roghayeh Tofigh, Sara Tutunchi, Ghodratollah Panahi, Fatemeh Borhani, Saeedeh Akhavan, Parisa Nourmohammadi, Sayyed M.H. Ghaderian, Milad Rasouli, Hamed Mirzaei.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2 Gastric and esophageal cancers are as main cancers of the gastrointestinal (GI) tract, which are associated with poor diagnosis and survival. Several efforts were made in the past few decades to finding effective therapeutic approaches, but these approaches had several problems. Finding new biomarkers is a critical step in finding new approaches for the treatment of these cancers. Finding new biomarkers that cover various aspects of the diseases could provide a choice of suitable therapies and better monitoring of patients with these cancers. Among several biomarkers tissue specific and circulating microRNAs (miRNAs) have emerged as powerful candidates in the diagnosis of gastric and esophageal cancers. MiRNAs are small noncoding single‐stranded RNA molecules that are found in the blood and regulate gene expression. These have numerous characteristics that make them suitable for being used as ideal biomarkers in cancer diagnosis. Research has indicated that the level and profile of miRNA in serum and plasma are very high. They are potentially noninvasive and sensitive enough to detect tumors in their primary stages of infection. Multiple lines of evidence indicate that the presence, absence, or deregulation of several circulating miRNAs (i.e., let‐7a, miR‐21, miR‐93, miR‐192a, miR‐18a, and miR‐10b for gastric cancer, and miR‐21, miR‐375, miR‐25‐3p, miR‐151a‐3p, and miR‐100‐3p for esophageal cancer) are associated with initiation and progression of gastric and esophageal cancers. The aim of this review is to highlight the recent advances in the roles of miRNAs in diagnosis and treatment of gastric and esophageal cancers. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8538-8550, November 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26850   open full text
• Transdifferentiation of human gingival mesenchymal stem cells into functional keratinocytes by Acalypha indica in three‐dimensional microenvironment.
  Dinesh Murugan Girija, Mangathayaru Kalachaveedu, Suresh Ranga Rao, Rajasekaran Subbarayan.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2 Gingival tissue is reportedly a promising, easily accessible, abundant resource of mesenchymal stem cells (MSC) for use in various tissue engineering strategies. Human gingival MSC (HGMSCs) were successfully isolated from gingival tissue and characterized. To analyze in a two‐dimensional form, HGMSCs were cultured with basal medium and induced with 25 µg/ml of Acalypha indica. Quantitative real‐time polymerase chain reaction (qPCR) and western blot analysis showed the presence of keratinocyte‐specific markers, including cytokeratin‐5 and involucrin. To further assess its capability for stratification akin to human keratinocytes, HGMSCs were encapsulated in a HyStem®‐HP Cell Culture Scaffold Kit and cultured in the presence of A. indica. Calcein AM staining indicated that the HyStem®‐HP Scaffold Kit has excellent biocompatibility. Immunofluorescence and qPCR analysis revealed the presence of keratinocyte‐specific markers. The study concluded that the three‐dimensional microenvironment is a novel method for inducing epidermal differentiation of HGMSCs to engineer epidermal substitutes with the help of A. indica, which provides an alternative strategy for skin tissue engineering. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8450-8457, November 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26807   open full text
• Promotion of the prehierarchical follicle growth by postovulatory follicles involving PGE2–EP2 signaling in chickens.
  Xin Lin, Xingting Liu, Changquan Guo, Mengqi Liu, Yuling Mi, Caiqiao Zhang.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2 The postovulatory follicle (POF) in birds is an enigmatic structure, the function of which remains largely unknown. Previous studies on chickens have shown that removal of POFs leads to the postponement of oviposition and the disturbance of broody behavior. One suggestion is that POFs may secrete some crucial hormones or cytokines to act on reproductive organs. However, such secretions and their specific target organs remain to be identified. Here, we investigate the putative functions of POFs in promoting the development of prehierarchical follicles in chickens and explore the possible signaling mechanisms controlling these processes. Results show that POFs express steroidogenic acute regulatory protein (STAR), cholesterol side‐chain cleavage enzyme (CYP11A1), cyclooxygenase 1 (COX1), and COX2 in granulosa cells (GCs), and, most notably, that POF1 produces more prostaglandin E2 (PGE2) or prostaglandin F2α than do the F1 follicle or the other POFs. Using coculture systems, we also found that POF1 or GCs from POF1 (POF1‐GCs) significantly promote the proliferation of theca externa cells of small white follicles (SWFs, one phase of the prehierarchical follicle). Treatment with PGE2 significantly facilitates theca externa cell proliferation in SWFs. This POF‐stimulating effect on SWF growth was prevented by treatment with indomethacin (COX inhibitor) or TG6‐10‐1 (PGE2 type 2 receptor [EP2] antagonist). Therefore, POF1 may secrete PGE2 to stimulate the progression of SWF by PGE2–EP2 signaling. These results indicate that POF1 may serve as a transient supplementary endocrine gland in the chicken ovary that stimulates the development of the prehierarchical follicles through PGE2–EP2 signaling. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8984-8995, November 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26844   open full text
• Establishment of an in vitro organoid model of dermal papilla of human hair follicle.
  Abhishak C. Gupta, Shikha Chawla, Ashok Hegde, Divya Singh, Balaji Bandyopadhyay, Chandrasekharan C. Lakshmanan, Gurpreet Kalsi, Sourabh Ghosh.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2 Human hair dermal papilla (DP) cells are specialized mesenchymal cells that play a pivotal role in hair regeneration and hair cycle activation. The current study aimed to first develop three‐dimensional (3D) DP spheroids (DPS) with or without a silk–gelatin (SG) microenvironment, which showed enhanced DP‐specific gene expression, resulting in enhanced extracellular matrix (ECM) production compared with a monolayer culture. We tested the feasibility of using this DPS model for drug screening by using minoxidil, which is a standard drug for androgenic alopecia. Minoxidil‐treated DPS showed enhanced expression of growth factors and ECM proteins. Further, an attempt has been made to establish an in vitro 3D organoid model consisting of DPS encapsulated by SG hydrogel and hair follicle (HF) keratinocytes and stem cells. This HF organoid model showed the importance of structural features, cell–cell interaction, and hypoxia akin to in vivo HF. The study helped to elucidate the molecular mechanisms to stimulate cell proliferation, cell viability, and elevated expression of HF markers as well as epithelial–mesenchymal crosstalks, demonstrating high relevance to human HF biology. This simple in vitro DP organoid model system has the potential to provide significant insights into the underlying mechanisms of HF morphogenesis, distinct molecular signals relevant to different stages of the hair cycle, and hence can be used for controlled evaluation of the efficacy of new drug molecules. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 9015-9030, November 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26853   open full text
• Extracellular vesicles derived from human embryonic stem cell‐MSCs ameliorate cirrhosis in thioacetamide‐induced chronic liver injury.
  Soura Mardpour, Seyedeh‐Nafiseh Hassani, Saeid Mardpour, Forough Sayahpour, Massoud Vosough, Jafar Ai, Nasser Aghdami, Amir Ali Hamidieh, Hossein Baharvand.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ Various somatic tissue‐derived mesenchymal stromal cells (MSCs) have been considered as an attractive therapeutic tool for treatment of liver diseases in which the secretion of soluble factors or extracellular vesicles (EVs) is the most probable mechanism. The experimental application of human embryonic stem cell‐derived MSC (ES‐MSC) increased rapidly and showed promising results, in vitro and in vivo. However, possible therapeutic effects of human ES‐MSC and their EVs on Thioacetamide (TAA)‐induced chronic liver injury have not been evaluated yet. Our data indicated that human ES‐MSC can significantly suppress the proliferation of peripheral blood mononuclear cells compared to bone marrow (BM)‐MSC and adipose (AD)‐MSC. Moreover, ES‐MSC increased the secretion of anti‐inflammatory cytokines (i.e., TGF‐β and IL‐10) and decreased IFN‐γ, compared to other MSCs. ES‐MSC EVs demonstrated immunomodulatory activities comparable to parental cells and ameliorated cirrhosis in TAA‐induced chronic rat liver injury, that is, reduction in fibrosis and collagen density, necrosis, caspase density, portal vein diameter, and transaminitis. The gene expression analyses also showed upregulation in collagenases (MMP9 and MMP13), anti‐apoptotic gene (BCL‐2) and anti‐inflammatory cytokines (TGF‐β1 and IL‐10) and down‐regulation of major contributors to fibrosis (Col1α, αSMA, and TIMP1), pro‐apoptotic gene (BAX) and pro‐inflammatory cytokines (TNFα and IL‐2) following treatment with ES‐MSC and ES‐MSC‐EV. These results demonstrated that human ES‐MSC and ES‐MSC EV as an off‐the‐shelf product, that needs further assessment to be suggested as an allogeneic product for therapeutic applications for liver fibrosis. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9330-9344, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26413   open full text
• Recombinant human hepatocyte growth factor provides protective effects in cerulein‐induced acute pancreatitis in mice.
  Mayrel Palestino‐Dominguez, Mario Pelaez‐Luna, Roberto Lazzarini‐Lechuga, Ignacio Rodriguez‐Ochoa, Veronica Souza, Roxana U. Miranda, Benjamín Perez‐Aguilar, Leticia Bucio, Jens U. Marquardt, Luis Enrique Gomez‐Quiroz, Maria Concepcion Gutierrez‐Ruiz.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ Acute pancreatitis is a multifactorial disease associated with profound changes of the pancreas induced by release of digestive enzymes that lead to increase in proinflammatory cytokine production, excessive tissue necrosis, edema, and bleeding. Elevated levels of hepatocyte growth factor (HGF) and its receptor c‐Met have been observed in different chronic and acute pancreatic diseases including experimental models of acute pancreatitis. In the present study, we investigated the protective effects induced by the recombinant human HGF in a mouse model of cerulein‐induced acute pancreatitis. Pancreatitis was induced by 8 hourly administrations of supramaximal cerulein injections (50 µg/kg, ip). HGF treatment (20 µg/kg, iv), significantly attenuated lipase content and amylase activity in serum as well as the degree inflammation and edema overall leading to less severe histologic changes such as necrosis, induced by cerulein. Protective effects of HGF were associated with activation of pro‐survival pathways such as Akt, Erk1/2, and Nrf2 and increase in executor survival‐related proteins and decrease in pro‐apoptotic proteins. In addition, ROS content and lipid peroxidation were diminished, and glutathione synthesis increased in pancreas. Systemic protection was observed by lung histology. In conclusion, our data indicate that HGF exerts an Nrf2 and glutathione‐mediated protective effect on acute pancreatitis reflected by a reduction in inflammation, edema, and oxidative stress. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9354-9364, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26444   open full text
• High folate intake contributes to the risk of large for gestational age birth and obesity in male offspring.
  Kaipeng Xie, Ziyi Fu, Hui Li, Xiaohong Gu, Zhiyong Cai, Pengfei Xu, Xianwei Cui, Lianghui You, Xing Wang, Lijun Zhu, Chenbo Ji, Xirong Guo.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ Folate supplementation is recommended before and during early pregnancy to prevent neural tube defects, but the effect of red blood cell (RBC) folate on large for gestational age (LGA) is still unknown. We performed a nested case‐control study including 542 LGA cases and 1,084 appropriate for gestational age (AGA) controls to examine the association of RBC folate concentrations with risk of LGA. Then, male offspring of dams fed basic folic acid (2 mg/kg, control) or 10‐fold folic acid (20 mg/kg, HFol) diet before and during pregnancy were used to explore the effect of high folate intake on birth weight and long‐term effects. We observed higher RBC folate concentrations in the cases compared to controls (p = 0.039). After adjustment for maternal age, BMI at enrollment, gestational weeks at enrollment, gestational weeks at delivery and infant gender, higher RBC folate levels were significantly associated with increased risk of LGA (Ptrend = 0.003). Interestingly, male offspring of HFol dams showed the higher birth weight, elevated levels of post loading blood glucose at 9 and 13 weeks post‐weaning and increased triglyceride (TG) and total cholesterol (TC) levels at 17 weeks post‐weaning. Furthermore, we observed that high folate intake increased the proliferation and differentiation of adipose cells. Our results suggest that maternal high folate intake confers the risk of LGA birth and accelerates the development of obesity in male offspring. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9383-9389, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26520   open full text
• A multi‐method evaluation of the effects of Inflammatory cytokines (IL‐1β, IFN‐γ, TNF‐α) on pancreatic β‐cells.
  Kaipeng Xie, Bo Xu, Yuqing Zhang, Minjian Chen, Yinwen Ji, Jie Wang, Zhenyao Huang, Kun Zhou, Yankai Xia, Wei Tang.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ We aimed to explore the effects of Inflammatory cytokines (IL‐1β, IFN‐γ, TNF‐α) on pancreatic β‐cells. CCK‐8 assay showed that the cell viability decreased after 24 hr treatment of TNF‐α, 48 hr of IFN‐γ, and 84 hr of IL‐1β. EdU assay illustrated that after 24 hr treatment, there were significantly reduced EdU‐labeled red fluorescence cells in TNF‐α group while not in IFN‐γ and IL‐1β groups. Flow Cytometry results displayed that TNF‐α and IFN‐γ groups increased apoptosis while IL‐1β group did not. Cell apoptosis results found that there was an increase in the S‐phase population of IL‐1β and TNF‐α groups, however, there was no significant difference in cell cycle between IFN‐γ group and the control. TEM images showed that there were reduction in the number of granules and mitochondria in IL‐1β and IFN‐γ groups, in particular paucity of insulin granules and mitochondria in TNF‐α group. Radioimmunoassay results presented that TNF‐α inhibited glucose‐induced insulin secretion, while there were no significant changes in IL‐1β and IFN‐γ groups when compared with the control. Metabolomic analysis found amino acid metabolism and Krebs cycle were the most robust altered metabolism pathways after inflammatory cytokines treatments. Overall, the altered amino acid metabolism and Krebs cycle metabolism might be important mechanisms of TNF‐α induced mouse pancreatic β‐cells dysfuction. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9375-9382, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26518   open full text
• ADAM17 participates in the protective effect of paeoniflorin on mouse brain microvascular endothelial cells.
  Haifang Wang, Shuhui Ma, Jing Li, Miaomiao Zhao, Xueping Huo, Jingying Sun, Lijun Sun, Jun Hu, Qinshe Liu.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ Paeoniflorin (PF), the most abundant active ingredient of traditional Chinese herbal medicine Paeoniae Radix, has been recognized as a potential neuroprotectant due to its remarkable efficacy on mitigating cerebral infarction and preventing the neurodegenerative diseases. However, the precise mechanisms of PF remain incompletely understood. In this study, we first provided evidence for the protective effect of PF on hydrogen peroxide‐induced injury on mouse brain microvascular endothelial bEnd.3 cells, and for transactivation of the epidermal growth factor receptor (EGFR) signal induced by PF, suggesting that EGFR transactivation might be involved in the beneficial role of PF. Next, by detecting the phosphorylation of a disintegrin and metalloprotease 17 (ADAM17) at Thr 735 and performing loss‐of‐function experiments with the ADAM17 inhibitor and ADAM 17‐siRNA, we showed that PF‐induced transactivation of EGFR and downstream ERKs and AKT signaling pathways were dependent on ADAM17. Furthermore, PF‐induced phosphorylation of ADAM17 and the EGFR transactivation were inhibited by the inhibitors of adenosine A1 receptor (A1R) or Src kinase that were applied to cells prior to PF treatment, implying the involvement of A1R, and Src in the activation of ADAM17. Finally, PF reduced the cell surface level of TNF‐receptor 1 (TNFR1) and increased the content of soluble TNFR1 (sTNFR1) in the culture media, indicating that PF might enhance the shedding of sTNFR1. Taken together, we conclude that A1R and Src‐dependent activation of ADAM17 participates in PF‐induced EGFR transactivation and TNFR1 shedding on mouse brain microvascular endothelial cells, which may contributes to the neuroprotective effects of PF. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9320-9329, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26308   open full text
• Effect of restriction vegan diet's on muscle mass, oxidative status, and myocytes differentiation: A pilot study.
  Daniela Vanacore, Giovanni Messina, Stefania Lama, Giuseppe Bitti, Pasqualina Ambrosio, Giancarlo Tenore, Antonietta Messina, Vincenzo Monda, Silvia Zappavigna, Mariarosaria Boccellino, Ettore Novellino, Marcellino Monda, Paola Stiuso.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ This study was conceived to evaluate the effects of three different diets on body composition, metabolic parameters, and serum oxidative status. We enrolled three groups of healthy men (omnivores, vegetarians, and vegans) with similar age, weight and BMI, and we observed a significant decrease in muscle mass index and lean body mass in vegan compared to vegetarian and omnivore groups, and higher serum homocysteine levels in vegetarians and vegans compared to omnivores. We studied whether serum from omnivore, vegetarian, and vegan subjects affected oxidative stress, growth and differentiation of both cardiomyoblast cell line H9c2 and H‐H9c2 (H9c2 treated with H2O2 to induce oxidative damage). We demonstrated that vegan sera treatment of both H9c2 and H‐H9c2 cells induced an increase of TBARS values and cell death and a decrease of free NO2− compared to vegetarian and omnivorous sera. Afterwards, we investigated the protective effects of vegan, vegetarian, and omnivore sera on the morphological changes induced by H2O2 in H9c2 cell line. We showed that the omnivorous sera had major antioxidant and differentiation properties compared to vegetarian and vegan sera. Finally, we evaluated the influence of the three different groups of sera on MAPKs pathway and our data suggested that ERK expression increased in H‐H9c2 cells treated with vegetarian and vegan sera and could promote cell death. The results obtained in this study demonstrated that restrictive vegan diet could not prevent the onset of metabolic and cardiovascular diseases nor protect by oxidative damage. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9345-9353, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26427   open full text
• Over‐expression of DEC1 inhibits myogenic differentiation by modulating MyoG activity in bovine satellite cell.
  Yongzhen Huang, Xinsheng Lai, Linyong Hu, Chuzhao Lei, Xianyong Lan, Chunlei Zhang, Yun Ma, Li Zheng, Yue‐Yu Bai, Fengpeng Lin, Hong Chen.
  Journal of Cellular Physiology. June 19, 2018
  --- - |2+ Differentiated embryo chondrocyte 1 (DEC1), a member of basic‐helix‐loop‐helix transcription factor Bhlhe40, also called stimulated by retinoic acid 13, STRA13, plays an important role in the regulation of adipogenesis, tumorigenesis, peripheral circadian output, response to hypoxia, and development of metabolic syndrome. Previous studies suggested that DEC1 was involved in skeletal muscle development; however, its precise role in myoblast differentiation has not been determined. In the present study, we showed that DEC1 expressed ubiquitously in different bovine tissues and was down‐regulated in differentiated bovine satellite cells. Expression of muscle specific transcription factors (Myf5, MyoD, MyoG, and MHC) was significantly down‐regulated when DEC1 was over‐expressed by both CoCl2‐simulated hypoxia and Adenovirus‐mediated transduction in bovine satellite cells. Consistent with that, promoter analyses via luciferase reporter assay also revealed that overexpression of bovine DEC1 could inhibit MyoG promoter activity. In conclusion, overexpression of DEC1 blocked myogenesis by inhibiting MyoG promoter activity in bovine. Our results provided a new mechanism for the muscle growth, which would contribute to increase cattle meat productivity. - Journal of Cellular Physiology, Volume 233, Issue 12, Page 9365-9374, December 2018.
  

  June 19, 2018   doi: 10.1002/jcp.26471   open full text
• Is the secret for a successful aging to keep track of cancer pathways?
  Donatella Tramontano, Francesca De Amicis.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2 A successful aging could be gained by life satisfaction, social functioning, or psychological resources and, definitely, by increasing resistance to diverse age‐related pathologies. Nowadays, cancer can be considered an age‐related disease since the incidence of most cancers increases with age, rising more rapidly beginning in midlife. Although adults with extended longevity are less likely to develop cancer, it is now emerging that aging and cancer share common molecular links, and thus targeting these mechanisms may be suitable to treat multiple disorders, for the prolonging of healthy aging. At present, one of the cornerstones of antiaging is hormone‐replacement therapy to treat diseases associated with a state of age‐related sex‐hormone deficiency in women and men; however, many studies question the relationship of hormone replacement to cancer recurrence. Here, we discuss signaling and metabolic molecular crossroad linking aging and cancer. This is useful to argue about the current knowledge of prolongevity and druggable targets and to motivate specific intervention strategies that could modify practices of the aging population, activating multiple longevity pathways but keeping track of cancer pathways, thereby potentially preserving health status. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8467-8476, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26825   open full text
• The key genes underlying pathophysiology association between the type 2‐diabetic and colorectal cancer.
  Wen‐Fang Peng, Feng Bai, Kan Shao, Li‐Sha Shen, Hui‐Hua Li, Shan Huang.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2+ Although diabetes mellitus (DM) is reported as an independent risk factor for colorectal cancer (CRC) in many researches, the underlying pathophysiology is still unclear. We investigated the differentially expressed genes (DEGs) for the diabetes and CRC to reveal the underlying pathophysiological association between the type 2‐diabetic (T2D) and CRC. Gene expression profiles for T2D (GSE55650), CRC (GSE8671), and Metformin treated cell lines (GSE67342) were downloaded from GEO database. The DEGs between T2D samples and their control samples were identified with t‐test and variance analysis. After cluster analysis and functional enrichment analysis, protein‐protein interaction (PPI) network was constructed to find potential genes for diabetes and CRC in Metformin's treatment. Totally, we identified 583 overlapped genes, 169 common DEGs, and 414 independent DEGs between T2D and CRC samples. The common genes contained 89 up‐regulated (DEGs1) and 80 down‐regulated genes (DEGs3); and independent DEGs contained 270 down‐regulated genes (DEGs4) in diabetes and 144 down‐regulated genes (DEGs2) in CRC. In enrichment analysis, the Ribosome pathway was significantly enriched by the independent DEGs. The common genes were mainly enriched in some inflammatory related pathways. Two target genes of Metformin were significantly interacted with six hub genes (HADHB, NDUFS3, TAF1, MYC, HNFF4A, and MAX) with significant changes in expression values (P < 0.05, t‐test). To summary, it is suggested that the six hub genes might play important roles in the process of Metformin treatment for diabetes and CRC. However, specific pathology remains to be further studied. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8551-8557, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26440   open full text
• Inactivation of Tp53 and Pten drives rapid development of pleural and peritoneal malignant mesotheliomas.
  Eleonora Sementino, Craig W. Menges, Yuwaraj Kadariya, Suraj Peri, Jinfei Xu, Zemin Liu, Richard G. Wilkes, Kathy Q. Cai, Frank J. Rauscher, Andres J. Klein‐Szanto, Joseph R. Testa.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2 Malignant mesothelioma (MM) is a therapy‐resistant cancer arising primarily from the lining of the pleural and peritoneal cavities. The most frequently altered genes in human MM are cyclin‐dependent kinase inhibitor 2A (CDKN2A), which encodes components of the p53 (p14ARF) and RB (p16INK4A) pathways, BRCA1‐associated protein 1 (BAP1), and neurofibromatosis 2 (NF2). Furthermore, the p53 gene (TP53) itself is mutated in ~15% of MMs. In many MMs, the PI3K–PTEN–AKT–mTOR signaling node is hyperactivated, which contributes to tumor cell survival and therapeutic resistance. Here, we demonstrate that the inactivation of both Tp53 and Pten in the mouse mesothelium is sufficient to rapidly drive aggressive MMs. PtenL/L;Tp53L/L mice injected intraperitoneally or intrapleurally with adenovirus‐expressing Cre recombinase developed high rates of peritoneal and pleural MMs (92% of mice with a median latency of 9.4 weeks and 56% of mice with a median latency of 19.3 weeks, respectively). MM cells from these mice showed consistent activation of Akt–mTor signaling, chromosome breakage or aneuploidy, and upregulation of Myc; occasional downregulation of Bap1 was also observed. Collectively, these findings suggest that when Pten and Tp53 are lost in combination in mesothelial cells, DNA damage is not adequately repaired and genomic instability is widespread, whereas the activation of Akt due to Pten loss protects genomically damaged cells from apoptosis, thereby increasing the likelihood of tumor formation. Additionally, the mining of an online dataset (The Cancer Genome Atlas) revealed codeletions of PTEN and TP53 and/or CDKN2A/p14ARF in ~25% of human MMs, indicating that cooperative losses of these genes contribute to the development of a significant proportion of these aggressive neoplasms and suggesting key target pathways for therapeutic intervention. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8952-8961, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26830   open full text
• Mesenchymal stromal cells from amniotic fluid are less prone to senescence compared to those obtained from bone marrow: An in vitro study.
  Nicola Alessio, Caterina Pipino, Domitilla Mandatori, Pamela Di Tomo, Angela Ferone, Marco Marchiso, Mariarosa A.B. Melone, Gianfranco Peluso, Assunta Pandolfi, Umberto Galderisi.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2 Mesenchymal stromal cells (MSCs) are considered to be an excellent source in regenerative medicine. They contain several cell subtypes, including multipotent stem cells. MSCs are of particular interest as they are currently being tested using cell and gene therapies for a number of human diseases. They represent a rare population in tissues; for this reason, they require, before being transplanted, an in vitro amplification. This process may induce replicative senescence, thus affecting differentiation and proliferative capacities. Increasing evidence suggests that MSCs from fetal tissues are significantly more plastic and grow faster than MSCs from bone marrow. Here, we compare amniotic fluid mesenchymal stromal cells (AF‐MSCs) and bone marrow mesenchymal stromal cells (BM‐MSCs) in terms of cell proliferation, surface markers, multidifferentiation potential, senescence, and DNA repair capacity. Our study shows that AF‐MSCs are less prone to senescence with respect to BM‐MSCs. Moreover, both cell models activate the same repair system after DNA damage, but AF‐MSCs are able to return to the basal condition more efficiently with respect to BM‐MSCs. Indeed, AF‐MSCs are better able to cope with genotoxic stress that may occur either during in vitro cultivation or following transplantation in patients. Our findings suggest that AF‐MSCs may represent a valid alternative to BM‐MSCs in regenerative medicine, and, of great relevance, the investigation of the mechanisms involved in DNA repair capacity of both AF‐MSCs and BM‐MSCs may pave the way to their rational use in the medical field. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8996-9006, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26845   open full text
• Lumichrome inhibits osteoclastogenesis and bone resorption through suppressing RANKL‐induced NFAT activation and calcium signaling.
  Chuan Liu, Zhen Cao, Wen Zhang, Jennifer Tickner, Heng Qiu, Chao Wang, Kai Chen, Ziyi Wang, Renxiang Tan, Shiwu Dong, Jiake Xu.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2 The dynamic balance between bone resorption and bone formation is crucial to maintain bone mass. Osteoclasts are key cells that perform bone resorption while osteoblasts and osteocytes function in bone formation. Osteoporosis, a bone metabolism disease characterized by bone loss and degradation of bone microstructure, occurs when osteoclastic bone resorption outstrips osteoblastic bone synthesis. The interaction between receptor activator of nuclear factor κB ligand (RANKL) and RANK on the surface of bone marrow macrophages promotes osteoclast differentiation and activation. In this study, we found that lumichrome, a photodegradation product of riboflavin, inhibits RANKL‐induced osteoclastogenesis and bone resorption as determined by tartrate‐resistant acid phosphatase staining, immunofluorescence, reverse transcription‐polymerase chain reaction, and western blot. Our results showed that lumichrome represses the expression of osteoclast marker genes, including cathepsin K (Ctsk) and Nfatc1. In addition, lumichrome suppressed RANKL‐induced calcium oscillations, NFATc1, NF‐κB, and MAPK signaling activation. Moreover, lumichrome promoted osteoblast differentiation at an early stage, as demonstrated by upregulated expression of osteoblast marker genes Alp, Runx2, and Col1a1. We also found that lumichrome reduces bone loss in ovariectomized mice by inhibiting osteoclastogenesis. In summary, our data suggest the potential of lumichrome as a therapeutic drug for osteolytic diseases. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8971-8983, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26841   open full text
• Sideroxylin (Callistemon lanceolatus) suppressed cell proliferation and increased apoptosis in ovarian cancer cells accompanied by mitochondrial dysfunction, the generation of reactive oxygen species, and an increase of lipid peroxidation.
  Sunwoo Park, Whasun Lim, Wonsik Jeong, Fuller W. Bazer, Dongho Lee, Gwonhwa Song.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2+ Sideroxylin is a C‐methylated flavone isolated from Callistemon lanceolatus and exerts antimicrobial activity against Staphylococcus aureus. However, the anticancer effects of sideroxylin and its intracellular signaling mechanisms have not yet been identified. Results of our study showed that sideroxylin decreased cell proliferation and increased apoptosis, causing DNA fragmentation, depolarization of the mitochondrial membrane, the generation of reactive oxygen species, and an increase of lipid peroxidation in ovarian cancer cells (ES2 and OV90 cells). Additionally, sideroxylin activated the phosphorylation of ERK1/2, JNK, P38, and MAPK proteins and the use of LY294002, U0126, SB203580, and SP600125 to block their phosphorylation, respectively, in ES2 and OV90 cells. Collectively, the results of present study indicated that sideroxylin was a novel therapeutic agent to combat the proliferation of ovarian cancer cells through the induction of mitochondrial dysfunction and the activation of PI3 K and MAPK signal transduction. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8597-8604, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26540   open full text
• LINC00968 functions as an oncogene in osteosarcoma by activating the PI3K/AKT/mTOR signaling.
  Gang Liu, Dongtang Yuan, Peng Sun, Weidong Liu, Peng‐Fei Wu, Huan Liu, Guang‐Yang Yu.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2+ Osteosarcoma is recognized as a malignant tumor in the skeletal system. Long non‐coding RNAs (lncRNAs) have been exhibited to play crucial roles in osteosarcoma development. Our current study focused on the biological effects and mechanism of LINC00968 in osteosarcoma pathogenesis. We observed that LINC00968 was dramatically elevated in osteosarcoma cells including U2OS, MG63, Saos‐2, SW1353, and 143‐B cells compared to human osteoblast cell line hFOB. Silence of LINC00968 inhibited osteosarcoma cell growth and proliferation in vitro. Reversely, overexpression of LINC00968 promoted osteosarcoma cell survival and cell colony formation ability in Saos‐2 and 143‐B cells. In addition, LINC00968 was able to induce osteosarcoma cell migration and invasion through up‐regulating MMP‐2 and MMP‐9 protein levels. The phosphoinosmde‐3‐kinase/Protein Kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway has been reported to participate in several cancer types. Here, in our study, we found that PI3K/AKT/mTOR pathway was involved in osteosarcoma progression. Knockdown of LINC00968 inactivated PI3K/AKT/mTOR signaling pathway in vitro. Subsequently, in vivo tumor xenografts were established using 143‐B cells to investigate whether LINC00968 can induce osteosarcoma development in vivo. Consistently, it was indicated that inhibition of LINC00968 significantly inhibited osteosarcoma progression in vivo. Taken these together, in our research, LINC00968 could be provided as a novel prognostic biomarker and therapeutic target in osteosarcoma diagnosis and treatment. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8639-8647, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26624   open full text
• MiR‐128‐1‐5p regulates tight junction induced by selenium deficiency via targeting cell adhesion molecule 1 in broilers vein endothelial cells.
  Tingru Pan, Xueyuan Hu, Tianqi Liu, Zhe Xu, Na Wan, Yiming Zhang, Shu Li.
  Journal of Cellular Physiology. June 15, 2018
  --- - |2 Vein endothelial cells (VECs) constitute an important barrier for macromolecules and circulating cells from the blood to the tissues, stabilizing the colloid osmotic pressure of the blood, regulating the vascular tone, and rapidly changing the intercellular connection, and maintaining normal physiological function. Tight junction has been discovered as an important structural basis of intercellular connection and may play a key role in intercellular connection injuries or vascular diseases and selenium (Se) deficiency symptoms. Hence, we replicated the Se‐deficient broilers model and detected the specific microRNA in response to Se‐deficient vein by using quantitative real time‐PCR (qRT‐PCR) analysis. Also, we selected miR‐128‐1‐5p based on differential expression in vein tissue and confirmed its target gene cell adhesion molecule 1 (CADM1) by the dual luciferase reporter assay and qRT‐PCR in VECs. We made the ectopic miR‐128‐1‐5p expression for the purpose of validating its function on tight junction. The result showed that miR‐128‐1‐5p and CADM1 were involved in the ZO‐1‐mediated tight junction, increased paracellular permeability, and arrested cell cycle. We presumed that miR‐128‐1‐5p and Se deficiency might trigger tight junction. Interestingly, miR‐128‐1‐5p inhibitor and fasudil in part hinder the destruction of the intercellular structure caused by Se deficiency. The miR‐128‐1‐5p/CADM1/tight junction axis provides a new avenue toward understanding the mechanism of Se deficiency, revealing a novel regulation model of tight junction injury in vascular diseases. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8802-8814, November 2018.
  

  June 15, 2018   doi: 10.1002/jcp.26794   open full text
• Long noncoding ribonucleic acid NKILA induces the endoplasmic reticulum stress/autophagy pathway and inhibits the nuclear factor‐k‐gene binding pathway in rats after intracerebral hemorrhage.
  Jiaoying Jia, Mingming Zhang, Qi Li, Qian Zhou, Yugang Jiang.
  Journal of Cellular Physiology. June 12, 2018
  --- - |2 Long noncoding RNAs (lncRNAs) have emerged as an important class of molecules that have been associated with brain function and neurological disease, but the expression profiles of lncRNAs after intracerebral hemorrhage (ICH) remain to be elucidated. In this study, we determined the expression pattern of nuclear factor‐k‐gene binding (NF‐kB) interacting lncRNA (NKILA) after ICH and examined its respective effects on the endoplasmic reticulum stress (ERS)/autophagy pathway, hippocampal neuron loss, and the NF‐kB pathway after type VII collagenase‐induced ICH in rats. The regulatory mechanisms of NKILA were investigated by an intraperitoneal injection of small interfering (siRNA) against NKILA into rats after ICH. NKILA inhibition mediated by siRNA against NKILA was shown to significantly reduce ERS and autophagy, activate the NF‐kB pathway, decrease neurological deficits, brain edema, and injury, and induce blood–brain barrier breakdown, further leading to hippocampal neuron loss and the production of inflammation cytokines. Taken together, the demonstration that NKILA induces the ERS/autophagy pathway and inhibits the NF‐kB pathway after ICH supports the concept that NKILA functions as a novel target that is required for the attenuation of brain injuries after ICH. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8839-8849, November 2018.
  

  June 12, 2018   doi: 10.1002/jcp.26798   open full text
• Calcium: A novel and efficient inducer of differentiation of adipose‐derived stem cells into neuron‐like cells.
  Farjam Goudarzi, Heidar Tayebinia, Jamshid Karimi, Elahe Habibitabar, Iraj Khodadadi.
  Journal of Cellular Physiology. June 05, 2018
  --- - |2 This study comparatively investigated the effectiveness of calcium and other well‐known inducers such as isobutylmethylxanthine (IBMX) and insulin in differentiating human adipose‐derived stem cells (ADSCs) into neuronal‐like cells. ADSCs were immunophenotyped and differentiated into neuron‐like cells with different combinations of calcium, IBMX, and insulin. Calcium mobilization across the membrane was determined. Differentiated cells were characterized by cell cycle profiling, staining of Nissl bodies, detecting the gene expression level of markers such as neuronal nuclear antigen (NeuN), microtubule associated protein 2 (MAP2), neuron‐specific enolase (NSE), doublecortin, synapsin I, glial fibrillary acidic protein (GFAP), and myelin basic protein (MBP) by quantitative real‐time polymerase chain reaction (quantitative real‐time polymerase chain reaction (qRT‐PCR) and protein level by the immunofluorescence technique. Treatment with Ca + IBMX + Ins induced neuronal appearance and projection of neurite‐like processes in the cells, accompanied with inhibition of proliferation and halt in the cell cycle. A significantly higher expression of MBP, GFAP, NeuN, NSE, synapsin 1, doublecortin, and MAP2 was detected in differentiated cells, confirming the advantages of Ca + IBMX + Ins to the other combinations of inducers. Here, we showed an efficient protocol for neuronal differentiation of ADSCs, and calcium fostered differentiation by augmenting the number of neuron‐like cells and instantaneous increase in the expression of neuronal markers. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8940-8951, November 2018.
  

  June 05, 2018   doi: 10.1002/jcp.26826   open full text
• Application of induced pluripotent stem cell and embryonic stem cell technology to the study of male infertility.
  Javad Amini Mahabadi, Hamed Sabzalipour, Hassan Hassani Bafrani, Seyed Mohammad Gheibi Hayat, Hossein Nikzad.
  Journal of Cellular Physiology. June 05, 2018
  --- - |2 Stem cells (SCs) are classes of undifferentiated biological cells existing only at the embryonic, fetal, and adult stages that can divide to produce specialized cell types during fetal development and remain in our bodies throughout life. The progression of regenerative and reproductive medicine owes the advancement of respective in vitro and in vivo biological science on the stem cell nature under appropriate conditions. The SCs are promising therapeutic tools to treat currently of infertility because of wide sources and high potency to differentiate. Nevertheless, no effective remedies are available to deal with severe infertility due to congenital or gonadotoxic stem cell deficiency in prepubertal childhood. Some recent solutions have been developed to address the severe fertility problems, including in vitro formation of germ cells from stem cells, induction of pluripotency from somatic cells, and production of patient‐specific pluripotent stem cells. There is a possibility of fertility restoration using the in vitro formation of germ cells from somatic cells. Accordingly, the present review aimed at studying the literature published on the medical application of stem cells in reproductive concerns. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8441-8449, November 2018.
  

  June 05, 2018   doi: 10.1002/jcp.26757   open full text
• Long noncoding RNA FAM83H‐AS1 exerts an oncogenic role in glioma through epigenetically silencing CDKN1A (p21).
  Yong‐Yan Bi, Gang Shen, Yong Quan, Wei Jiang, Fulin Xu.
  Journal of Cellular Physiology. June 05, 2018
  --- - |2 Gliomas are the commonest and most aggressive primary malignant tumor in the central nervous system. Long noncoding RNAs (lncRNAs) have been identified to act as crucial regulators in multiple biological processes, including tumorigenesis. FAM83H antisense RNA1 (FAM83H‐AS1) has been uncovered to be dysregulated in several cancers. However, the biological role of FAM83H‐AS1 in glioma still needs to be investigated. Currently, our findings indicated that FAM83H‐AS1 was upregulated in glioma tissues and cell lines and high level of FAM83H‐AS1 was associated with poor prognosis of glioma. Loss‐of‐function assays demonstrated that silenced FAM83H‐AS1 obviously suppressed cell proliferation via regulating the cell‐cycle distribution and cell apoptosis rate, and mechanistic experiments revealed that FAM83H‐AS1 could epidemically silence CDKN1A expression through recruiting EZH2 to the promoter of CDKN1A, thereby influencing the cell cycle and proliferation. Collectively, our findings suggested that FAM83H‐AS1 participated in the progression of glioma and might act as a potential therapeutic target and prognosis biomarker for human glioma. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8896-8907, November 2018.
  

  June 05, 2018   doi: 10.1002/jcp.26813   open full text
• ARHGAP21 as a master regulator of multiple cellular processes.
  Lucas R. O. Rosa, Gabriela M. Soares, Leonardo R. Silveira, Antonio C. Boschero, Helena C. L. Barbosa‐Sampaio.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 The cellular cytoskeleton is involved with multiple biological processes and is tightly regulated by multiple proteins and effectors. Among these, the RhoGTPases family is one of the most important players. RhoGTPAses are, in turn, regulated by many other elements. In the past decade, one of those regulators, the RhoGAP Rho GTPase Activating Protein 21 (ARHGAP21), has been overlooked, despite being implied as having an important role on many of those processes. In this paper, we aimed to review the available literature regarding ARHGAP21 to highlight its importance and the mechanisms of action that have been found so far for this still unknown protein involved with cell adhesion, migration, Golgi regulation, cell trafficking, and even insulin secretion. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8477-8481, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26829   open full text
• A minireview of E4BP4/NFIL3 in heart failure.
  Bharath Kumar Velmurugan, Ruey‐Lin Chang, Shibu Marthandam Asokan, Chih‐Fen Chang, Cecilia‐Hsuan Day, Yueh‐Min Lin, Yuan‐Chuan Lin, Wei‐Wen Kuo, Chih‐Yang Huang.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Heart failure (HF) remains a major cause of morbidity and mortality worldwide. The primary cause identified for HF is impaired left ventricular myocardial function, and clinical manifestations may lead to severe conditions like pulmonary congestion, splanchnic congestion, and peripheral edema. Development of new therapeutic strategies remains the need of the hour for controlling the problem of HF worldwide. Deeper insights into the molecular mechanisms involved in etiopathology of HF indicate the significant role of calcium signaling, autocrine signaling pathways, and insulin‐like growth factor‐1 signaling that regulates the physiologic functions of heart growth and development such as contraction, metabolism, hypertrophy, cytokine signaling, and apoptosis. In view of these facts, a transcription factor (TF) regulating the myriad of these signaling pathways may prove as a lead candidate for development of therapeutics. Adenovirus E4 promoter‐binding protein (E4BP4), also known as nuclear‐factor, interleukin 3 regulated (NFIL3), a type of basic leucine zipper TF, is known to regulate the signaling processes involved in the functioning of heart. The current review discusses about the expression, structure, and functional role of E4BP4 in signaling processes with emphasis on calcium signaling mechanisms, autocrine signaling, and insulin‐like growth factor II receptor–mediated processes regulated by E4BP4 that may regulate the pathogenesis of HF. We propose that E4BP4, being the critical component for the regulation of the above signaling processes, may serve as a novel therapeutic target for HF, and scientific investigations are merited in this direction. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8458-8466, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26790   open full text
• Isoform‐specific effects of transforming growth factor β on endothelial‐to‐mesenchymal transition.
  Harika Sabbineni, Arti Verma , Payaningal R. Somanath.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Endothelial‐to‐mesenchymal transition (EndMT) was first reported in the embryogenesis. Recent studies show that EndMT also occurs in the disease progression of atherosclerosis, cardiac and pulmonary fibrosis, pulmonary hypertension, diabetic nephropathy, and cancer. Although transforming growth factor β (TGFβ) is crucial for EndMT, it is not clear which isoform elicits a predominant effect. The current study aims to directly compare the dose‐dependent effects of TGFβ1, TGFβ2, and TGFβ3 on EndMT and characterize the underlying mechanisms. In our results, all three TGFβ isoforms induced EndMT in human microvascular endothelial cells after 72 hr, as evidenced by the increased expression of mesenchymal markers N‐cadherin and α‐smooth muscle actin as well as the decreased expression of endothelial nitric oxide synthase. Interestingly, the effect of TGFβ2 was the most pronounced. At 1 ng/ml, only TGFβ2 treatment resulted in significantly increased phosphorylation (activation) of Smad2/3 and p38‐MAPK and increased expression of mesenchymal transcription factors Snail and FoxC2. Intriguingly, we observed that treatment with 1 ng/ml TGFβ1 and TGFβ3, but not TGFβ2, resulted in an increased expression of TGFβ2, thus indicating that EndMT with TGFβ1 and TGFβ3 treatments was due to the secondary effects through TGFβ2 secretion. Furthermore, silencing TGFβ2 using small interfering RNA blunted the expression of EndMT markers in TGFβ1‐ and TGFβ3‐treated cells. Together, our results indicate that TGFβ2 is the most potent inducer of EndMT and that TGFβ1‐ and TGFβ3‐induced EndMT necessitates a paracrine loop involving TGFβ2. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8418-8428, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26801   open full text
• Time‐dependent contribution of BMP, FGF, IGF, and HH signaling to the proliferation of mesenchymal stroma cells during chondrogenesis.
  Jennifer Fischer, Natalie Knoch, Tanja Sims, Nils Rosshirt, Wiltrud Richter.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Early loss of up to 50% of cells is common for in vitro chondrogenesis of mesenchymal stromal cells (MSC) in pellet culture, reducing the efficacy and the tissue yield for cartilage engineering. Enhanced proliferation could compensate for this unwanted effect, but relevant signaling pathways remain largely unknown. The aim of this study was to identify the contribution of bone morphogenetic protein (BMP), fibroblast growth factor (FGF), insulin‐like growth factor (IGF), and hedgehog (HH) signaling toward cell proliferation during chondrogenesis and investigate whether a further mitogenic stimulation is possible and promising. Human MSC were subjected to chondrogenesis in the presence or absence of pathway inhibitors or activators up to Day 14 or from Days 14 to 28, before proliferation, DNA and proteoglycan content were quantified. [3H]‐thymidine incorporation revealed arrest of proliferation on Day 3, after which cell division was reinitiated. Although BMP signaling was essential for proliferation throughout chondrogenesis, IGF signaling was relevant only up to Day 14. In contrast, FGF and HH signaling drove proliferation only from Day 14 onward. Early BMP4, IGF‐1, or FGF18 treatment neither prevented early cell loss nor allowed further mitogenic stimulation. However, application of the HH‐agonist purmorphamine from Day 14 increased proliferation 1.44‐fold (p < 0.05) and late BMP4‐application enhanced the DNA and proteoglycan content, with significant effects on tissue yield. Conclusively, a differential and phase‐dependent contribution of the four pathways toward proliferation was uncovered and BMP4 treatment was promising to enhance tissue yield. Culture forms less prone to size limitations by nutrient/oxygen gradients and a focus on early apoptosis prevention may be considered as the next steps to further enhance chondrocyte formation from MSC. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8962-8970, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26832   open full text
• SIRT1 inhibits TGF‐β‐induced endothelial‐mesenchymal transition in human endothelial cells with Smad4 deacetylation.
  Zhen Li, Fei Wang, Siyuan Zha, Qing Cao, Jing Sheng, Shuyan Chen.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Endothelial‐mesenchymal transition (EndMT) plays a pivotal role in organ fibrosis. This study examined the effect of SIRT1 on transforming growth factor beta (TGF‐β)‐induced EndMT in human endothelial cells (ECs) and its probable molecular mechanism. We assessed EndMT by immunofluorescence staining, quantitative real‐time polymerase chain reaction, Western blotting, and migration and invasion assays. Adenovirus was used to overexpress or knockdown SIRT1 in ECs. The regulatory relationship between SIRT1 and Smad4 was analyzed by coimmunoprecipitation assay. We found that SIRT1 was decreased in TGF‐β‐induced EndMT, and SIRT1 inhibited TGF‐β‐induced EndMT through deacetylating Smad4. Our findings suggest that SIRT1 has an important role in inhibiting EndMT by regulating the TGF‐β/Smad4 pathway in human ECs and, thus, protecting against fibrosis. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 9007-9014, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26846   open full text
• PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy.
  Xu Wang, Yang Li, Yan Fan, Xinmiao Yu, Xiaoyun Mao, Feng Jin.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Invasion and migration is the hallmark of malignant tumors as well as the major cause for breast cancer death. The polypyrimidine tract binding, PTB, protein serves as an important model for understanding how RNA binding proteins affect proliferation and invasion and how changes in the expression of these proteins can control complex programs of tumorigenesis. We have investigated some roles of polypyrimidine tract binding protein 1 (PTBP1) in human breast cancer. We found that PTBP1 was upregulated in breast cancer tissues compared with normal tissues and the same result was confirmed in breast cancer cell lines. Knockdown of PTBP1 substantially inhibited tumor cell growth, migration, and invasion. These results suggest that PTBP1 is associated with breast tumorigenesis and appears to be required for tumor cell growth and maintenance of metastasis. We further analyzed the relationship between PTBP1 and clinicopathological parameters and found that PTBP1 was correlated with her‐2 expression, lymph node metastasis, and pathological stage. This will be a novel target for her‐2(+) breast cancer. PTBP1 exerts these effects, in part, by regulating the phosphatase and tensin homolog‐phosphatidylinositol‐4,5‐bisphosphate 3‐kinase/protein kinase B (PTEN‐PI3K/Akt) pathway and autophagy, and consequently alters cell growth and contributes to the invasion and metastasis. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8930-8939, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26823   open full text
• Triose‐phosphate isomerase is a novel target of miR‐22 and miR‐28, with implications in tumorigenesis.
  Saife Niaz Lone, Raihana Maqbool, Fazl Q. Parray, Mahboob Ul Hussain.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Aerobic glycolysis is the hallmark of many cancer cells that results in a high rate of adenosine triphosphate (ATP) production and, more importantly, biosynthetic intermediates, which are required by the fast‐growing tumor cells. The molecular mechanism responsible for the increased glycolytic influx of tumor cells is still not fully understood. In the present study, we have attempted to address the above question by exploring the role of the glycolytic enzyme, triose‐phosphate isomerase (TPI), in the cancer cells. The western blot analysis of the 30 human colorectal cancer samples depicted higher post‐transcriptional expression of TPI in the tumor tissue relative to the normal tissue. In addition, we identified two novel microRNAs, miR‐22 and miR‐28, that target the TPI messenger RNA (mRNA) and regulate its expression. miR‐22 and the miR‐28 showed significant inverse expression status viz‐a‐viz the expression of the TPI. The specificity of the miR‐22/28 regulation of the TPI mRNA was confirmed by various biochemical and mutagenic assays. Moreover, the hypoxia conditions resulted in an increased expression of the TPI protein, with a concomitant decrease in miR‐22/28. The physiological significance of the TPI and miR‐22/28 interaction for the glycolytic influx was confirmed by the l‐lactate production in the HCT‐116+/+ cells. Overall, our data demonstrate the novel microRNA mediated post‐transcriptional regulation of the TPI glycolytic enzyme, which may be one of the possible reasons for the increased glycolytic capacity of the tumor cells. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8919-8929, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26821   open full text
• In vivo antivascular endothelial growth factor treatment induces corneal endothelium apoptosis in rabbits through changes in p75NTR–proNGF pathway.
  Magda Gharbiya, Alice Bruscolini, Marta Sacchetti, Pamela Rosso, Valentina Carito, Marco Segatto, Elena Fico, Paola Tirassa, Alessandro Lambiase.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Intravitreal injection (IVT) of antivascular endothelial growth factor (anti‐VEGF) agents is widely used for the treatment of retinal vascular diseases. Recently, the injection of anti‐VEGF agents in the ocular anterior chamber has been proposed for the treatment of neovascular glaucoma and potential side effects on the corneal structures have been investigated with contrasting results. Increasing evidence has demonstrated that VEGF inhibition is associated with cellular apoptotic changes and that this effect may be mediated by alterations in nerve growth factor (NGF) pathway. In this study, we demonstrated that anterior chamber injection (IC), but not IVT injection of two different anti‐VEGF agents, aflibercept and ranibizumab, affects rabbit corneal endothelium in terms of survival and apoptosis and is associated with changes in endothelial expression of NGF precursor (proNGF) and p75 neurotrophin receptor (p75NTR) receptor. We observed an increase in corneal endothelial cell incorporation of trypan blue and expression of cleaved‐caspase 3 (c‐Casp3), p75NTR, and RhoA after IC injection of both anti‐VEGF drugs when compared with the vehicle. Our results showed that apoptosis induction by aflibercept was more pronounced when compared with that of ranibizumab. Aflibercept also mediated a significant increase in endothelial expression of proNGF when compared with the vehicle. In line with these data, IC administration of both anti‐VEGF agents induced the activation of apoptotic signals in endothelial cells, including an increase in c‐Casp3, decrease in Bad Ser 112 phosphorylation, and unbalance of AKT phosphorylation. These results demonstrated that administration of anti‐VEGF in the anterior chamber of rabbit affects endothelial cell survival by inducing apoptosis through alteration of NGF pathway. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8874-8883, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26806   open full text
• MALAT1 via microRNA‐17 regulation of insulin transcription is involved in the dysfunction of pancreatic β‐cells induced by cigarette smoke extract.
  Qian Sun, Hui Xu, Junchao Xue, Qianlei Yang, Chao Chen, Ping Yang, Aohan Han, Qingyun Tu, Jiachun Lu, Xiaohua Gao, Quanyong Xiang, Qizhan Liu.
  Journal of Cellular Physiology. June 01, 2018
  --- - |2 Cigarettes contain various chemicals with the potential to influence metabolic health. Exposure to cigarette smoke causes a dysfunction in pancreatic β‐cells and impairs insulin production. However, the mechanisms for cigarette smoke‐induced reduction of insulin remain largely unclear. Data from 558 patients with diabetes showed that, with smoking pack‐years, homeostatic model assessment (HOMA)‐β (a method for assessing β‐cell function) decreased and that HOMA of insulin resistance increased. For β‐cells (MIN6), cigarette smoke extract (CSE) increased the levels of thioredoxin‐interacting protein (TXNIP) and the long noncoding (lnc)RNA, metastasis‐associated lung adenocarcinoma transcript 1 (MALAT1), and downregulated the levels of the transcription factor, mafA, and microRNA (miR)‐17. MALAT1, one of four lncRNAs predicted to regulate miR‐17, was knocked down by small interfering RNA (siRNA). For these cells, an miR‐17 mimic inhibited TXNIP and enhanced the production of insulin. Knockdown of MALAT1 induced an increase in miR‐17, which suppressed TXNIP and promoted the production of insulin. In the sera of patients with diabetes who smoked, there were higher MALAT1 levels and lower miR‐17 levels than in the sera of nonsmokers. Thus, CSE inhibits insulin production by upregulating TXNIP via MALAT1‐mediated downregulation of miR‐17, which provides an understanding of the processes involved in the reduced β‐cells function caused by cigarette smoke. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8862-8873, November 2018.
  

  June 01, 2018   doi: 10.1002/jcp.26800   open full text
• Elevated transforming growth factor β signaling activation in β‐actin‐knockout mouse embryonic fibroblasts enhances myofibroblast features.
  Xin Xie, Piergiorgio Percipalle.
  Journal of Cellular Physiology. May 31, 2018
  --- - |2 Signaling by the transforming growth factor‐β (TGF‐β) is an essential pathway regulating a variety of cellular events. TGF‐β is produced as a latent protein complex and is required to be activated before activating the receptor. The mechanical force at the cell surface is believed to be a mechanism for latent TGF‐β activation. Using β‐actin null mouse embryonic fibroblasts as a model, in which actin cytoskeleton and cell‐surface biophysical features are dramatically altered, we reveal increased TGF‐β1 activation and the upregulation of TGF‐β target genes. In β‐actin null cells, we show evidence that the enhanced TGF‐β signaling relies on the active utilization of latent TGF‐β1 in the cell culture medium. TGF‐β signaling activation contributes to the elevated reactive oxygen species production, which is likely mediated by the upregulation of Nox4. The previously observed myofibroblast phenotype of β‐actin null cells is inhibited by TGF‐β signaling inhibition, while the expression of actin cytoskeleton genes and angiogenic phenotype are not affected. Together, our study shows a scenario that the alteration of the actin cytoskeleton and the consequent changes in cellular biophysical features lead to changes in cell signaling process such as TGF‐β activation, which in turn contributes to the enhanced myofibroblast phenotype. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8884-8895, November 2018.
  

  May 31, 2018   doi: 10.1002/jcp.26808   open full text
• Two controls of cell proliferation underlie cancer relapse.
  Arthur B. Pardee, Chiang J. Li.
  Journal of Cellular Physiology. May 31, 2018
  --- - |2+ Much progress has been made in understanding the basis of cancer. Current therapies can effectively shrink tumors. But they frequently relapse, metastasize to other locations, and are lethal. Effective therapies are very much needed for preventing this relapse. Creation of a eukaryotic organism commences with one original stem cell, a fertilized egg, which multiplies and differentiates. Mutations of normal stem cells can produce cancer stem cells (CSC). These cells may resist chemotherapy, proliferate, and produce new tumors. Human chorionic gonadotrophin (hCG) is composed of two proteins (alpha and beta) that bind to the cell membrane and activate a number of intracellular pathways. hCG has been shown to activate the proliferation of cancer stem cells. Cyclin dependent regulation of the adult cells is created in normal differentiation and replaces the hCG regulation of stem cells. To selectively kill the cancer stem cells conventional cancer therapies could be followed with a therapy based on inactivating human chronic gonadotrophin (HCG). For example chemically modified prostaglandins like RU486 prevent binding of the unmodified steroid to hCG and inactivate hCG. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8437-8440, November 2018.
  

  May 31, 2018   doi: 10.1002/jcp.26597   open full text
• The imbalance of circulating T helper subsets and regulatory T cells in patients with LRBA deficiency: Correlation with disease severity.
  Gholamreza Azizi, Abbas Mirshafiey, Hassan Abolhassani, Reza Yazdani, Alireza Ghanavatinejad, Farshid Noorbakhsh, Nima Rezaei, Asghar Aghamohammadi.
  Journal of Cellular Physiology. May 28, 2018
  --- - |2 Patients with lipopolysaccharides responsive beige‐like anchor protein (LRBA) deficiency suffer from a variety of immunological abnormalities. In the current study, we investigated the role of T helper (Th) cell subsets and regulatory T (Treg) cells and their related cytokines and transcription factors in the immune dysregulation of LRBA deficiency. The study population comprised of 13 LRBA‐deficient patients and 13 age‐ and sex‐matched healthy controls (HCs). Th subsets and Treg were examined by flow cytometry. The expression of determinant cytokines (interferon‐γ [IFN‐γ], interleukin [IL]‐17, IL‐22, and IL‐10), and cell subset–specific transcription factors were evaluated before and after proliferation and activation stimuli. The frequencies of Th1, Th1‐like Th17 and Th22 cells along with the expression of T‐box transcription factor (TBET) and runt‐related transcription factor 1 (RUNX1) were significantly increased in patients with LRBA. Moreover, IFN‐γ and IL‐22 production in LRBA‐deficient CD4+ T cells were elevated after lymphocyte stimulation, particularly in patients with enteropathy. However, CD4+CD25+FoxP3+CD127− cells were significantly decreased in LRBA‐deficient patients compared with those of HCs, particularly in patients with autoimmunity. There was a negative correlation between the frequencies of CD4+CD25+FoxP3+CD127− cells and Th1‐like Th17 cells in LRBA‐deficient patients, and an overlapping phenotype of autoimmunity and enteropathy were observed in ~70% of patients. The frequency of Th17 cells was lower in patients with enteropathy, while Th1‐like Th17 cells were higher than in those without enteropathy. Our findings demonstrated an imbalance in Th subsets, mainly in Th1‐like Th17 and Treg cells and their corresponding cytokines in LRBA deficiency, which might be important in the immunopathogenesis of autoimmunity and enteropathy. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8767-8777, November 2018.
  

  May 28, 2018   doi: 10.1002/jcp.26772   open full text
• Interaction of CacyBP/SIP with NPM1 and its influence on NPM1 localization and function in oxidative stress.
  Sara Rosińska, Anna Filipek.
  Journal of Cellular Physiology. May 28, 2018
  --- - |2 Calcyclin (S100A6) binding protein/Siah‐1 interacting protein (CacyBP/SIP) is mainly a cytoplasmic protein; however, some literature data suggested its presence in the nucleus. In this work we examined more precisely the nuclear localization and function of CacyBP/SIP. By applying mass spectrometry, we have identified several nuclear proteins, among them is nucleophosmin (NPM1), that may interact with CacyBP/SIP. Subsequent assays revealed that CacyBP/SIP forms complexes with NPM1 in the cell and that the interaction between these two proteins is direct. Interestingly, although CacyBP/SIP exhibits phosphatase activity, we have found that its overexpression favors phosphorylation of NPM1 on S125. In turn, the RNA immunoprecipitation assay indicated that the altered CacyBP/SIP level has an impact on the amount of 28S and 18S rRNA bound to NPM1. The overexpression of CacyBP/SIP resulted in a significant increase in the binding of 28S and 18S rRNA to NPM1, whereas silencing of CacyBP/SIP expression decreased 28S rRNA binding and had no effect on the binding of 18S rRNA. Further studies have shown that under oxidative stress, CacyBP/SIP overexpression alters NPM1 distribution in cell nuclei. In addition, staining for a nucleolar marker, fibrillarin, revealed that CacyBP/SIP is indispensable for maintaining the nucleolar structure. These results are in agreement with data obtained by western blot analysis, which show that upon oxidative stress the NPM1 level decreases but that CacyBP/SIP overexpression counteracts the effect of stress. Altogether, our results show for the first time that CacyBP/SIP binds to and affects the properties of a nuclear protein, NPM1, and that it is indispensable for preserving the structure of nucleoli under oxidative stress. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8826-8838, November 2018.
  

  May 28, 2018   doi: 10.1002/jcp.26797   open full text
• Key genes and functional coexpression modules involved in the pathogenesis of systemic lupus erythematosus.
  Shushan Yan, Weijie Wang, Guohong Gao, Min Cheng, Xiaodong Wang, Zengyan Wang, Xiufen Ma, Chunxiang Chai, Donghua Xu.
  Journal of Cellular Physiology. May 28, 2018
  --- - |2 We performed a systematic review of genome‐wide gene expression datasets to identify key genes and functional modules involved in the pathogenesis of systemic lupus erythematosus (SLE) at a systems level. Genome‐wide gene expression datasets involving SLE patients were searched in Gene Expression Omnibus and ArrayExpress databases. Robust rank aggregation (RRA) analysis was used to integrate those public datasets and identify key genes associated with SLE. The weighted gene coexpression network analysis (WGCNA) was adapted to identify functional modules involved in SLE pathogenesis, and the gene ontology enrichment analysis was utilized to explore their functions. The aberrant expressions of several randomly selected key genes were further validated in SLE patients through quantitative real‐time polymerase chain reaction. Fifteen genome‐wide gene expression datasets were finally included, which involved a total of 1,778 SLE patients and 408 healthy controls. A large number of significantly upregulated or downregulated genes were identified through RRA analysis, and some of those genes were novel SLE gene signatures and their molecular roles in etiology of SLE remained vague. WGCNA further successfully identified six main functional modules involved in the pathogenesis of SLE. The most important functional module involved in SLE included 182 genes and mainly enriched in biological processes, including defense response to virus, interferon signaling pathway, and cytokine‐mediated signaling pathway. This study identifies a number of key genes and functional coexpression modules involved in SLE, which provides deepening insights into the molecular mechanism of SLE at a systems level and also provides some promising therapeutic targets. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8815-8825, November 2018.
  

  May 28, 2018   doi: 10.1002/jcp.26795   open full text
• The promise of stem cell markers in the diagnosis and therapy of epithelial dysplasia and oral squamous cell carcinoma.
  Farnaz Mohajertehran, Amirhossein Sahebkar, Reza Zare, Nooshin Mohtasham.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Oral squamous cell carcinoma (OSCC) is the most common type of head and neck cancer. Epithelial dysplasia is often initiated in the cells and cell nuclei adjacent to the epithelial cell membrane. Reduced cell–cell adhesions enable cancer cells to detach from the tumor and disseminate to other organs. The mutations in epithelial dysplasia markers such as E‐cadherin and epithelial cell adhesion molecules (CD326) can lead to proliferation, growth and survival of the tumor cells and persistence of numerous malignancies that play a key role in epithelial dysplasia of OSCC. Accordingly, these genes can be considered prognostic markers or potential therapeutic targets for the tailored management of patients with OSCC. The gene expression profile of OSCC stem cells indicates a differential pattern that facilitates establishing a cell signature. Owing to the highly tumorigenic behavior of cancer stem cells and the role of these cells in tumor differentiation, treatment resistance, relapse, and metastasis, we reviewed the role of stem cell markers in epithelial dysplasia and OSCC. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8499-8507, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26789   open full text
• Mcl‐1 targeting could be an intriguing perspective to cure cancer.
  Anna De Blasio, Renza Vento, Riccardo Di Fiore.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 The Bcl‐2 family, which plays important roles in controlling cancer development, is divided into antiapoptotic and proapoptotic members. The change in the balance between these members governs the life and death of the cells. Mcl‐1 is an antiapoptotic member of this family and its distribution in normal and cancerous tissues strongly differs from that of Bcl‐2. In human cancers, where upregulation of antiapoptotic proteins is common, Mcl‐1 expression is regulated independent of Bcl‐2 and its inhibition promotes senescence, a major barrier to tumorigenesis. Cancer chemotherapy determines various kinds of responses, such as senescence and autophagy; however, the ideal response to chemotherapy is apoptosis. Mcl‐1 is a potent oncogene that is regulated at the transcriptional, posttranscriptional, and posttranslational levels. Mcl‐1 is a short‐lived protein that, in the NH2 terminal region, contains sites for posttranslational regulation that can lead to proteasomal degradation. The USP9X Mcl‐1 deubiquitinase regulates Mcl‐1 and the levels of these two proteins are strongly correlated. Mcl‐1 has three splicing variants (the antiapoptotic protein Mcl‐1L and the proapoptotic proteins Mcl‐1S and Mcl‐1ES), each contributing toward apoptosis regulation. In cancers responsible for the most deaths in the world, the presence of Mcl‐1 is associated with malignant cell growth and evasion of apoptosis. Mcl‐1 is also one of the key regulators of cancer stem cells’ self‐renewal that contributes to tumor survival. A great number of indirect and selective Mcl‐1 inhibitors have been produced and some of these have shown efficacy in several clinical trials. Thus, therapeutic manipulation of Mcl‐1 can be a useful strategy to combat cancer. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8482-8498, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26786   open full text
• Bone marrow mesenchymal stem cell donors with a high body mass index display elevated endoplasmic reticulum stress and are functionally impaired.
  Baris Ulum, Hikmet Taner Teker, Aysun Sarikaya, Gunay Balta, Baris Kuskonmaz, Duygu Uckan‐Cetinkaya, Fatima Aerts‐Kaya.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Bone marrow mesenchymal stem cells (BM‐MSCs) are promising candidates for regenerative medicine purposes. The effect of obesity on the function of BM‐MSCs is currently unknown. Here, we assessed how obesity affects the function of BM‐MSCs and the role of endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) therein. BM‐MSCs were obtained from healthy donors with a normal (<25) or high (>30) body mass index (BMI). High‐BMI BM‐MSCs displayed severely impaired osteogenic and diminished adipogenic differentiation, decreased proliferation rates, increased senescence, and elevated expression of ER stress–related genes ATF4 and CHOP. Suppression of ER stress using tauroursodeoxycholic acid (TUDCA) and 4‐phenylbutyrate (4‐PBA) resulted in partial recovery of osteogenic differentiation capacity, with a significant increase in the expression of ALPL and improvement in the UPR. These data indicate that BMI is important during the selection of BM‐MSC donors for regenerative medicine purposes and that application of high‐BMI BM‐MSCs with TUDCA or 4‐PBA may improve stem cell function. However, whether this improvement can be translated into an in vivo clinical advantage remains to be assessed. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8429-8436, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26804   open full text
• Chronic phosphodiesterase type 5 inhibition has beneficial effects on subcutaneous adipose tissue plasticity in type 2 diabetic mice.
  Daniela Fiore, Daniele Gianfrilli, Silvia Cardarelli, Fabio Naro, Andrea Lenzi, Andrea M. Isidori, Mary A. Venneri.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Different adipose tissue (AT) depots are associated with multiple metabolic risks. Phosphodiesterase type 5 (PDE5) is involved in adipocyte physiology and PDE5 inhibition may affect adipogenesis and ameliorate white AT quality. The aim of this study is to investigate the distribution of AT and the composition of the stroma‐vascular fraction (SVF) of subcutaneous AT (SAT) in type 2 diabetic mice after prolonged treatment with a PDE5 inhibitor, Sildenafil. 18 db/db mice were treated with Sildenafil or vehicle for 12 weeks. AT distribution was monitored and SAT was processed for isolation of SVF by flow cytometry. Sildenafil induced an overall reduction in AT, mainly in visceral AT (VAT), compared with SAT. In Sildenafil‐treated mice, the mean change in body weight from baseline positively correlated with VAT, but not with SAT. Characterization of SVF of SAT showed an increase in the frequency of M2 macrophages and endothelial cells in treated mice. Sildenafil improved the maintenance of SAT homeostasis and distribution. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8411-8417, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26796   open full text
• Zafirlukast promotes insulin secretion by increasing calcium influx through L‐type calcium channels.
  Hyeon‐Jeong Hwang, Kyoung‐Su Park, Jang Hyun Choi, Lucio Cocco, Hyun‐Jun Jang, Pann‐Ghill Suh.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2+ The zafirlukast has been reported to be anti‐inflammatory and widely used to alleviate the symptoms of asthma. However, its influence on insulin secretion in pancreatic β‐cells has not been investigated. Herein, we examined the effects of zafirlukast on insulin secretion and the potential underlying mechanisms. Among the cysteinyl leukotriene receptor 1 antagonists, zafirlukast, pranlukast, and montelukast, only zafirlukast enhanced insulin secretion in a concentration‐dependent manner in both low and high glucose conditions and elevated the level of [Ca2+]i, further activating Ca2+/calmodulin‐dependent protein kinase II (CaMKII), protein kinase B (AKT), and extracellular signal‐regulated kinase (ERK) signaling. These effects were nearly abolished by the L‐type Ca2+ channel antagonist nifedipine, while treatment with thapsigargin, a sarco/endoplasmic reticulum Ca2+ ATPase inhibitor, did not have the same effect, suggesting that zafirlukast primarily induces the entry of extracellular Ca2+ rather than intracellular Ca2+ from the endoplasmic reticulum. Zafirlukast treatment resulting in a significant drop in glucose levels and increased insulin secretion in C57BL/6J mice. These findings will contribute to an improved understanding of the side effects of zafirlukast and potential candidate for a therapeutic intervention in diabetes. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8701-8710, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26750   open full text
• Excessive training induces molecular signs of pathologic cardiac hypertrophy.
  Alisson L. da Rocha, Giovana R. Teixeira, Ana P. Pinto, Gustavo P. Morais, Luciana da C. Oliveira, Larissa Gaioto de Vicente, Lilian E. C. M. da Silva, José R. Pauli, Dennys E. Cintra, Eduardo R. Ropelle, Leandro P. de Moura, Rania A. Mekary, Ellen C. de Freitas, Adelino S. R. da Silva.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Chronic exercise induces cardiac remodeling that promotes left ventricular hypertrophy and cardiac functional improvement, which are mediated by the mammalian or the mechanistic target of rapamycin (mTOR) as well as by the androgen and glucocorticoid receptors (GRs). However, pathological conditions (i.e., chronic heart failure, hypertension, and aortic stenosis, etc.) also induce cardiac hypertrophy, but with detrimental function, high levels of proinflammatory cytokines and myostatin, elevated fibrosis, reduced adenosine monophosphate‐activated protein kinase (AMPK) activation, and fetal gene reactivation. Furthermore, recent studies have evidenced that excessive training induced an inflammatory status in the serum, muscle, hypothalamus, and liver, suggesting a pathological condition that could also be detrimental to cardiac tissue. Here, we verified the effects of three running overtraining (OT) models on the molecular parameters related to physiological and pathological cardiac hypertrophy. C57BL/6 mice performed three different OT protocols and were evaluated for molecular parameters related to physiological and pathological cardiac hypertrophy, including immunoblotting, reverse transcription polymerase chain reaction, histology, and immunohistochemistry analyses. In summary, the three OT protocols induced left ventricle (LV) hypertrophy with signs of cardiac fibrosis and negative morphological adaptations. These maladaptations were accompanied by reductions in AMPKalpha (Thr172) phosphorylation, androgen receptor, and GR expressions, as well as by an increase in interleukin‐6 expression. Specifically, the downhill running–based OT model reduced the content of some proteins related to the mTOR signaling pathway and upregulated the β‐isoform of myosin heavy‐chain gene expression, presenting signs of LV pathological hypertrophy development. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8850-8861, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26799   open full text
• Long non‐coding RNA FEZF1‐AS1 promotes breast cancer stemness and tumorigenesis via targeting miR‐30a/Nanog axis.
  Zhi Zhang, Liwei Sun, Yixuan Zhang, Guanming Lu, Yongqiang Li, Zhongheng Wei.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2+ Long non‐coding RNAs (lncRNAs) have been verified to modulate the tumorigenesis of breast cancer at multiple levels. In present study, we aim to investigate the role of lncRNA FEZF1‐AS1 on breast cancer‐stem like cells (BCSC) and the potential regulatory mechanism. In breast cancer tissue, lncRNA FEZF1‐AS1 was up‐regulated compared with controls and indicated poor prognosis of breast cancer patients. In vitro experiments, FEZF1‐AS1 was significantly over‐expressed in breast cancer cells, especially in sphere subpopulation compared with parental subpopulation. Loss‐of‐functional indicated that, in BCSC cells (MDA‐MB‐231 CSC, MCF‐7 CSC), FEZF1‐AS1 knockdown reduced the CD44+/CD24− rate, the mammosphere‐forming ability, stem factors (Nanog, Oct4, SOX2), and inhibited the proliferation, migration and invasion. In vivo, FEZF1‐AS1 knockdown inhibited the breast cancer cells growth. Bioinformatics analysis tools and series of validation experiments confirmed that FEZF1‐AS1 modulated BCSC and Nanog expression through sponging miR‐30a, suggesting the regulation of FEZF1‐AS1/miR‐30a/Nanog. In summary, our study validate the important role of FEZF1‐AS1/miR‐30a/Nanog in breast cancer stemness and tumorigenesis, providing a novel insight and treatment strategy for breast cancer. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8630-8638, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26611   open full text
• TANK‐binding kinase 1 and Janus kinase 2 play important roles in the regulation of mitogen‐activated protein kinase phosphatase‐1 expression after toll‐like receptor 4 activation.
  Eunji Kim, Ju Y. Yoon, Jongsung Lee, Deok Jeong, Jae G. Park, Yo H. Hong, Ji H. Kim, Adithan Aravinthan, Jong‐Hoon Kim, Jae Y. Cho.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Inflammation is a response that protects the body from pathogens. Through several inflammatory signaling pathways mediated by various families of transcription factors, such as nuclear factor‐κB (NF‐κB), activator protein‐1 (AP‐1), interferon regulatory factors (IRFs), and signal transducers and activators of transcription (STATs), various inflammatory cytokines and chemokines are induced and inflammatory responses are boosted. Simultaneously, inhibitory systems are activated and provide negative feedback. A typical mechanism by which this process occurs is that inflammatory signaling molecules upregulate mitogen‐activated protein kinase phosphatase‐1 (MKP1) expression. Here, we investigated how kinases regulate MKP1 expression in lipopolysaccharide‐triggered cascades. We found that p38 and c‐Jun N‐terminal kinase (JNK) inhibitors decreased MKP1 expression. Using specific inhibitors, gene knockouts, and gene knockdowns, we also found that tumor necrosis factor receptor‐associated factor family member‐associated nuclear factor κB activator (TANK)‐binding kinase 1 (TBK1) and Janus kinase 2 (JAK2) are involved in the induction of MKP1 expression. By analyzing JAK2‐induced activation of STATs, STAT3‐specific inhibitors, promoter binding sites, and STAT3−/− cells, we found that STAT3 is directly linked to TBK1‐mediated and JAK2‐mediated induction of MKP1 expression. Our data suggest that MKP1 expression can be differentially regulated by p38, JNK, and the TBK1–JAK2–STAT3 pathway after activation of toll‐like receptor 4 (TLR4). These data also imply crosstalk between the AP‐1 pathway and the IRF3 and STAT3 pathways. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8790-8801, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26787   open full text
• Spheroids from adipose‐derived stem cells exhibit an miRNA profile of highly undifferentiated cells.
  A. Barbara Di Stefano, Federica Grisafi, Marta Castiglia, Alessandro Perez, Luigi Montesano, Alessandro Gulino, Francesca Toia, Daniele Fanale, Antonio Russo, Francesco Moschella, Angelo A. Leto Barone, Adriana Cordova.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Two‐dimensional (2D) cell cultures have been extensively used to investigate stem cell biology, but new insights show that the 2D model may not properly represent the potential of the tissue of origin. Conversely, three‐dimensional cultures exhibit protein expression patterns and intercellular junctions that are more representative of their in vivo condition. Multiclonal cells that grow in suspension are defined as “spheroids,” and we have previously demonstrated that spheroids from adipose‐derived stem cells (S‐ASCs) displayed enhanced regenerative capability. With the current study, we further characterized S‐ASCs to further understand the molecular mechanisms underlying their stemness properties. Recent studies have shown that microRNAs (miRNAs) are involved in many cellular mechanisms, including stemness maintenance and proliferation, and adipose stem cell differentiation. Most studies have been conducted to identify a specific miRNA profile on adherent adipose stem cells, although little is still known about S‐ASCs. In this study, we investigate for the first time the miRNA expression pattern in S‐ASCs compared to that of ASCs, demonstrating that cell lines cultured in suspension show a typical miRNA expression profile that is closer to the one reported in induced pluripotent stem cells. Moreover, we have analyzed miRNAs that are specifically involved in two distinct moments of each differentiation, namely early and late stages of osteogenic, adipogenic, and chondrogenic lineages during long‐term in vitro culture. The data reported in the current study suggest that S‐ASCs have superior stemness features than the ASCs and they represent the true upstream stem cell fraction present in adipose tissue, relegating their adherent counterparts. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8778-8789, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26785   open full text
• NEAT1 upregulates TGF‐β1 to induce hepatocellular carcinoma progression by sponging hsa‐mir‐139‐5p.
  Jianfei Tu, Zhongwei Zhao, Min Xu, Xiaojie Lu, Liu Chang, Jiansong Ji.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2+ Increasing evidence has shown that the lncRNA Nuclear Enriched Abundant Transcript 1 (NEAT1) play important roles in cell proliferation, migration, and invasion in various tumors. In our current study, we concentrated on the biological mechanisms of NEAT1 in hepatocellular carcinoma (HCC) development. It was found that NEAT1 was significantly increased in human HCC cell lines including Hep3B, LM3, MHCC97L, SK‐hep1, and HepG2 cells compared to the normal human liver cell line LO2. Meanwhile, we observed that hsa‐miR‐139‐5p was greatly decreased in HCC cells, which suggested a negative correlation between NEAT1 and hsa‐mir‐139‐5p. In addition, NEAT1 downregulation can restrain HCC cell growth, migration, and invasion. Consistently, overexpression of hsa‐mir‐139‐5p exerted a similar phenomenon. Dual‐luciferase reporter assay, RIP assay, and RNA pull‐down assay confirmed that NEAT1 can function as a ceRNA by sponging hsa‐mir‐139‐5p. In addition, TGF‐β1 was identified as a downstream target of hsa‐mir‐139‐5p and hsa‐mir‐139‐5p overexpression was able to suppress TGF‐β1 levels. Furthermore, it was indicated that TGF‐β1 inhibition can inhibit HCC cell growth, migration, and invasion ability. Taken these together, we speculated that NEAT1 can modulate TGF‐β1 expression by sponging hsa‐mir‐139‐5p in HCC. These data indicates that targeting the NEAT1/hsa‐mir‐139‐5p/TGF‐β1 axis could be a new strategy for HCC. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8578-8587, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26524   open full text
• The immunomodulatory effects of adipose‐derived mesenchymal stem cells and mesenchymal stem cells‐conditioned medium in chronic colitis.
  Maryam Heidari, Sedigheh Pouya, Kaveh Baghaei, Hamid Asadzadeh Aghdaei, Saeed Namaki, Mohammad Reza Zali, Seyed Mahmoud Hashemi.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Inflammatory bowel disease (IBD) as a chronic recurrent disorder is characterized by mucosal immune response dysregulation, which is more prevalent in the youth. Adipose‐derived mesenchymal stem cells (ADMSCs) are the multipotent cells that can be effective in immune response regulation via cell–cell interaction and their secretions. In this study, the effects of ADMSCs and mesenchymal stem cell‐conditioned medium (MSC‐CM) were evaluated on dextran sulfate sodium (DSS)‐induced colitis in mice. Chronic colitis was induced in female C57BL/6 mice using 2% DSS in drinking water for three cycles; there were 4 days of DSS‐water administration that was followed by 7 days of DSS‐free water, in a cycle. ADMSCs, 106 cells per mouse, were injected intraperitoneally (IP), whereas the MSC‐CM injection was also performed six times from the last day of DSS in Cycle 1. Clinical symptoms were recorded daily. The colon pathological changes, cytokine levels, and regulatory T (Treg) cell percentages were then analyzed. After receiving ADMSCs and MSC‐CM in colitis mice, the clinical symptoms and disease activity index were improved and the survival rate was increased. The histopathological examination also showed tissue healing in comparison with the nontreated group. In addition, the increased level of transforming growth factor beta, increased percentage of Treg cells, increased level of interleukin (IL)‐10, and decreased level of IL‐17 were observed after the treatment. This study showed the regulatory effects of ADMSCs and MSC‐CM on inflammatory responses. Therefore, the use of ADMSCs and MSC‐CM can be introduced as a new and effective therapeutic approach for patients with colitis. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8754-8766, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26765   open full text
• Umbelliprenin shows antitumor, antiangiogenesis, antimetastatic, anti‐inflammatory, and immunostimulatory activities in 4T1 tumor‐bearing Balb/c mice.
  Mohsen Rashidi, Ahad Khalilnezhad, Davar Amani, Hamidreza Jamshidi, Ahad Muhammadnejad, Ali Bazi, Seyed Ali Ziai.
  Journal of Cellular Physiology. May 24, 2018
  --- - |2 Umbelliprenin (UMB) has shown various pharmacological properties in vitro. We investigated the antineoplastic and immunostimulatory effects of UMB in 4T1 mammary‐tumor‐bearing mice. Two‐hundred microliter of UMB (12.5 mg/ml) was intraperitoneally administrated to healthy and tumor‐bearing female Balb/c mice for a period of 18 days. Data was analyzed using GraphPad Prism 5 software for Windows (version 5, La Jolla, CA). UMB caused a significant decrease in tumor size (P < 0.01). Serum interferon gamma (IFNγ) was augmented in both healthy and tumor‐bearing animals (P < 0.01), and IL‐4 declined in healthy animals (P < 0.01) treated with UMB. Expressions of Ki‐67, VEGF, CD31, MMP2, MMP9, VCAM1, and NF‐κB were significantly decreased in tumors from UMB‐treated animals (P < 0.001), whereas E‐Cadherin and TNFR1 expressions were markedly increased (P < 0.001). The rates of liver and lung metastases in UMB‐administrated animals were smaller compared to the control. UMB can potently inhibit tumor growth, angiogenesis, metastasis, and inflammation and potentiate an antitumor immune response in vivo. However, further investigations are required to evaluate the UMB mechanisms of action in cancerous cells. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8908-8918, November 2018.
  

  May 24, 2018   doi: 10.1002/jcp.26814   open full text
• Articular cartilage protection in Ctsk‐/‐ mice is associated with cellular and molecular changes in subchondral bone and cartilage matrix.
  Fabiana N. Soki, Ryu Yoshida, David N. Paglia, Le T. Duong, Marc F. Hansen, Hicham Drissi.
  Journal of Cellular Physiology. May 21, 2018
  --- - |2+ Osteoarthritis (OA) is a degenerative disease and a major cause of chronic disability in aging individuals. Cathepsin K (CatK), encoded by the Ctsk gene, has been implicated in the pathogenesis of pycnodysostosis and osteoporosis. The use of a selective inhibitor of CatK was recently shown to delay OA progression in rabbits. However, the cellular mechanisms underlying these protective effects remain unexplored. We examined articular cartilage maintenance and joint bone remodeling using Ctsk null mice (Ctsk‐/‐) which underwent destabilization of the medial meniscus (DMM). We found that Ctsk‐/‐ mice displayed delayed remodeling of subchondral and calcified cartilage by osteoclasts and chodroclasts respectively in DMM‐induced osteoarthritis. While WT mice displayed a more severe OA phenotype than Ctsk‐/‐ mice at 16 weeks, higher subchondral bone volume and lower trabecular spacing were also observed in surgically‐induced OA joints of Ctsk‐/‐ mice. However, no differences were seen in non‐surgical controls. During OA progression, TRAP+ osteoclast numbers were increased in both WT and Ctsk‐/‐ mice. However, Ctsk‐/‐ mice had fewer physis‐derived chondroclasts than WT when OA was present. These data suggest that CatK may differentially regulate chondroclastogenesis in the growth plate. Targeted PCR arrays of RNA harvested from laser captured osteoclasts in the subchondral bone and chondroclasts in the growth plate demonstrated differential expression of Atp6v0d2, Tnfrsf11a, Ca2, Calcr, Ccr1, Gpr68, Itgb3, Nfatc1, and Syk genes between WT and Ctsk‐/‐ mice at 8‐ and 16‐weeks post‐DMM. Our data provide insight into the cellular mechanisms by which cathepsin K deletion delays OA progression in mice. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8666-8676, November 2018.
  

  May 21, 2018   doi: 10.1002/jcp.26745   open full text
• Effect of estradiol on fibroblasts from postmenopausal idiopathic carpal tunnel syndrome patients.
  Yoshiaki Yamanaka, Kunitaka Menuki, Takafumi Tajima, Yasuaki Okada, Kenji Kosugi, Yukichi Zenke, Akinori Sakai.
  Journal of Cellular Physiology. May 21, 2018
  --- - |2+ Fibrosis of the subsynovial connective tissue (SSCT) is a characteristic finding in patients with idiopathic carpal tunnel syndrome (CTS). Idiopathic CTS frequently occurs in postmenopausal women; therefore, female steroid hormones, especially estrogens, may be involved in its development. In this study, we evaluated the effect of the estradiol on the expression of genes and proteins related to fibrosis of SSCT fibroblasts from patients with idiopathic CTS. This study included 10 postmenopausal women (mean age 76 years). Fibroblasts derived from SSCT were treated with estradiol (10−4–10−12 M), and the expression levels of TGF‐β‐responsive genes were evaluated. The relationships between the expression of untreated estrogen receptor α (ERα) and ERβ and changes in gene expression due to estradiol treatment were examined by quantitative real‐time polymerase chain reaction. The effects of 10−4 M estradiol on collagen type I (Col1) and collagen type III (Col3) protein expression levels were also evaluated by fluorescent staining. The relationships between ERα/β and Col1/3 expression were evaluated by immunohistochemical staining. The reduction in Col1A1 mRNA expression due to estradiol treatment was positively correlated with ERα expression (r = 0.903, p < 0.01). At the protein level, expression of Col1 and Col3 were down‐regulated. These results indicated that ERα‐mediated signaling may be involved in the regulation of Col1A1, and its regulatory effect may be dependent on the ERα expression level. The accurate evaluation of ERα expression level in the SSCT of individual patients with idiopathic CTS might guide the effective use of new estrogen replacement therapy. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8723-8730, November 2018.
  

  May 21, 2018   doi: 10.1002/jcp.26752   open full text
• Effects of δ‐tocotrienol on ochratoxin A—induced nephrotoxicity in rats.
  Sara Damiano, Luigi Navas, Patrizia Lombari, Serena Montagnaro, Iris M. Forte, Antonio Giordano, Salvatore Florio, Roberto Ciarcia.
  Journal of Cellular Physiology. May 18, 2018
  --- - |2+ Ochratoxin A (OTA), is a natural contaminant of the food chain worldwide involved in the development of different type of cancers in animals and humans. Several studies suggested that oxidative damage might contribute to increase the cytotoxicity and carcinogenicity capabilities of OTA. The aim of this study was to evaluate the possible protective effect of δ‐tocotrienol (Delta), a natural form of vitamin E, against OTA‐induced nephrotoxicity. Male Sprague–Dawley rats were treated with OTA and/or Delta by gavage for 14 days. Our results shown that OTA treatment induced the increase of reactive oxigen species production correlated to a strong reduction of Glomerular Filtration Rate (GFR) and absoluted fluid reabsorption (Jv) with conseguent significant increase in blood pressure. Consistent, we noted in the kidney of rats treated with OTA, an increase in malondialdheyde and dihydroethidium production and a reduction of the activity of the catalase, superoxide dismutase, and glutathione peroxidase. Conversly, in the rat group subjected to the concomitant treatment OTA plus Delta, we observed the restored effect, compared the OTA treatment group, on blood pressure, GFR, Jv, and all activities of renal antioxidant enzymes. Finally, as far as concern the tissue damage induced by OTA and measured evaluating fibronectin protein levels, we observed that in OTA plus Delta group this effect is not restored. Our findings releval that a mechanism underlying the renal toxicity induced by OTA is the oxidative stress and provide a new rationale to use a Delta in order to protect, at least in part, against OTA‐induced nephrotoxicity. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8731-8739, November 2018.
  

  May 18, 2018   doi: 10.1002/jcp.26753   open full text
• Let‐7e inhibits TNF‐α expression by targeting the methyl transferase EZH2 in DENV2‐infected THP‐1 cells.
  Yingke Zhang, Qianqian Zhang, Lian Gui, Yan Cai, Xiaohong Deng, Cheukfai Li, Qi Guo, Xiaoshun He, Junqi Huang.
  Journal of Cellular Physiology. May 16, 2018
  --- - |2+ Tumor necrosis factor α (TNFα), an important inflammatory cytokine, is associated with dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS), a severe pathological manifestation of dengue virus (DENV) infection. However, the regulatory mechanism of microRNA on TNFα is currently unknown. Our study showed that the TNFα expression increased immediately and then later decreased, while a marked increase for the miRNA let‐7e was detected in dengue virus type 2 (DENV2)‐infected peripheral blood mononuclear cells (PBMCs). From this study, we found that let‐7e was able to inhibit TNFα expression, but bioinformatics analysis showed that the enhancer of zeste homolog 2 (EZH2) was the potential direct target of let‐7e instead of TNFα. EZH2 methyl transferase can produce H3K27me3 and has a negative regulatory role. Using a dual‐luciferase reporter assay and Western blotting, we confirmed that EZH2 was a direct target of let‐7e and found that siEZH2 could inhibit TNFα expression. In the further study of the regulatory mechanism of EZH2 on TNFα expression, we showed that siEZH2 promoted EZH1 and H3K4me3 expression and inhibited H3K27me3 expression. More importantly, we revealed that siEZH2 down‐regulated NF‐κB p65 within the nucleus. These findings indicate that the let‐7e/EZH2/H3K27me3/NF‐κB p65 pathway is a novel regulatory axis of TNFα expression. In addition, we determined the protein differences between siEZH2 and siEZH2‐NC by iTRAQ and found a number of proteins that might be associated with TNFα. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8605-8616, November 2018.
  

  May 16, 2018   doi: 10.1002/jcp.26576   open full text
• Inhibitory effects of tubeimoside I on synoviocytes and collagen‐induced arthritis in rats.
  Zhenzhou Liu, Lin Zhou, Xuemei Ma, Shengnan Sun, Haiwen Qiu, Hui Li, Jiake Xu, Mei Liu.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Advancements in rheumatoid arthritis (RA) therapies have shown considerable progresses in the comprehension of disease. However, the development of new potential medicines with relative safety and efficacy continues and natural compounds have been considered as alternatives or complementary agents to gain immense attractions. Tubeimoside I (TBMS I), a main triterpenoid saponin isolated from Bolbostemma paniculatum, has been reported to possess antiviral and anticancer effects. However, its effect on RA remains unknown. Here, we investigated the therapeutic effect of TBMS I in collagen‐induced arthritis (CIA) rats and explored its underlying mechanism. Our results showed that TBMS I treatment efficaciously ameliorated inflammation and joint destruction of rats with CIA. In vitro studies revealed that TBMS I suppressed the production of pro‐inflammatory cytokines including IL‐1β, IL‐6, IL‐8 and TNFα, and downregulated the expression of MMP‐9. In addition, TBMS I attenuated the destructive phenotypes of FLS of CIA rats including inhibiting proliferation and reducing migration rate. Further mechanistic analysis demonstrated that TBMS I suppressed TNFα‐induced activations of NF‐κB and MAPKs (p38 and JNK) leading to the downregulation of pro‐inflammatory cytokines, which was beneficial to the anti‐proliferative and anti‐migratory activities of FLS cells. Taken together, TBMS I has a great potential to be developed into a novel therapeutic agent for the treatment of RA. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8740-8753, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26754   open full text
• Multiple phosphorylation sites on γ‐tubulin are essential and contribute to the biogenesis of basal bodies in Tetrahymena.
  Ewa Joachimiak, Maria Jerka‐Dziadosz, Łucja Krzemień‐Ojak, Ewa Wacławek, Katarzyna Jedynak, Paulina Urbanska, Wojciech Brutkowski, Hanna Sas‐Nowosielska, Hanna Fabczak, Jacek Gaertig, Dorota Wloga.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ The mechanisms that regulate γ‐tubulin, including its post‐translational modifications, are poorly understood. γ‐Tubulin is important for the duplication of centrioles and structurally similar basal bodies (BBs), organelles which contain a ring of nine triplet microtubules. The ciliate Tetrahymena thermophila carries hundreds of cilia in a single cell and provides an excellent model to specifically address the role of γ‐tubulin in the BBs assembly and maintenance. The genome of Tetrahymena contains a single γ‐tubulin gene. We show here that there are multiple isoforms of γ‐tubulin that are likely generated by post‐translational modifications. We identified evolutionarily conserved serine and threonine residues as potential phosphosites of γ‐tubulin, including S80, S129, S131, T283, and S360. Several mutations that either prevent (S80A, S131A, T283A, S360A) or mimic (T283D) phosphorylation were conditionally lethal and at a higher temperature phenocopied a loss of γ‐tubulin. Cells that overproduced S360D γ‐tubulin displayed phenotypes consistent with defects in the microtubule‐dependent functions, including an asymmetric division of the macronucleus and abnormalities in the pattern of BB rows, including gaps, fragmentation, and misalignment. In contrast, overexpression of S129D γ‐tubulin affected the orientation, docking, and structure of the BBs, including a loss of either the B‐ or C‐subfibers or the entire triplets. We conclude that conserved potentially phosphorylated amino acids of γ‐tubulin are important for either the assembly or stability of BBs. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8648-8665, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26742   open full text
• NEAT1 contributes to breast cancer progression through modulating miR‐448 and ZEB1.
  Xing Jiang, Yong Zhou, Ai‐Jun Sun, Jun‐Li Xue.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Breast cancer is a kind of common female cancers. Increasing evidence has exhibited that lncRNAs exert a crucial role in breast cancer. So far, the mechanism of lncRNAs in breast cancer is still not well established. In our current study, we focused on the biological role of lncRNA Nuclear Enriched Abundant Transcript 1 (NEAT1) in breast cancer. We observed that NEAT1 levels were significantly increased in human breast cancer cells including MCF‐7, MDA‐MB‐453, MDA‐MB‐231, and SKBR3 cells compared to normal mammary epithelial cells MCF‐10A while miR‐448 was decreased. We found that downregulation of NEAT1 was able to inhibit the growth of breast cancer cells and miR‐448 mimic exerted the similar function. Bioinformatics analysis and dual luciferase reporter assays confirmed the negative correlation between NEAT1 and miR‐448 in vitro. In addition, ZEB1 was predicted as a novel mRNA target of miR‐448. Overexpression of NEAT1 can induce breast cancer cell growth, migration, and invasion by inhibiting miR‐448 and upregulating ZEB1. It was demonstrated that NEAT1 can increase ZEB1 levels while miR‐448 mimic can repress ZEB1. It was speculated in our study that NEAT1 can serve as a competing endogenous lncRNA (ceRNA) to modulate ZEB1 by sponging miR‐448 in breast cancer. To conclude, we uncovered that NEAT1 participated in breast cancer progression by regulating miR‐448 and ZEB1. NEAT1 can be provided as a vital biomarker in breast cancer diagnosis and treatment therapy. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8558-8566, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26470   open full text
• Zinc mitigates renal ischemia‐reperfusion injury in rats by modulating oxidative stress, endoplasmic reticulum stress, and autophagy.
  Najet Hadj Abdallah, Anna Baulies, Ahlem Bouhlel, Mohamed Bejaoui, Mohamed A. Zaouali, Safa Ben Mimouna, Imed Messaoudi, José C. Fernandez‐Checa, Carmen García Ruiz, Hassen Ben Abdennebi.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Oxidative stress is a major factor involved in the pathogenesis of renal ischemia/reperfusion (I/R). Exogenous zinc (Zn) was suggested as a potent antioxidant; however, the mechanism by which it strengthens the organ resistance against the effects of reactive oxygen species (ROS) is not yet investigated. The present study aims to determine whether acute zinc chloride (ZnCl2) administration could attenuate endoplasmic reticulum (ER) stress, autophagy, and inflammation after renal I/R. Rats were subjected to either sham operation (Sham group, n = 6), or 1 hr of bilateral ischemia followed by 2 hr of reperfusion (I/R groups, n = 6), or they received ZnCl2 orally 24 hr and 30 min before ischemia (ZnCl2 group, n = 6). Rats were subjected to 1 hr of bilateral renal ischemia followed by 2 hr of reperfusion (I/R group, n = 6). Our results showed that ZnCl2 enhances renal function and reduces cytolysis (p < 0,05). In addition, it increased significantly the activities of antioxidant enzymes (SOD, CAT, and GPX) and the level of GSH in comparison to I/R (p < 0,05). Interestingly, ZnCl2 treatment resulted in significant decreased ER stress, as reflected by GRP78, ATF‐6,p‐eIF‐2α, XPB‐1, and CHOP downregulaion. Rats undergoing ZnCl2 treatment demonstrated a low expression of autophagy parameters (Beclin‐1 and LAMP‐2), which was correlated with low induction of apoptosis (caspase‐9, caspase‐3, and p‐JNK), and reduction of inflammation (IL‐1ß, IL‐6, and MCP‐1) (p < 0,05). In conclusion, we demonstrated the potential effect of Zn supplementation to modulate ER pathway and autophagic process after I/R. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8677-8690, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26747   open full text
• Downregulated expression of microRNA‐329 inhibits apoptosis of nigral dopaminergic neurons by regulating CDKN2D expression via the FoxO3a signaling pathway in rats with Parkinson's disease.
  Yuan‐Yuan Liu, Yi‐Nan Zhang, Qing‐Shan Yang.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Parkinson's disease (PD) is a common neurodegenerative disorder due to the loss of dopaminergic neurons in the substantia nigra. This study focuses on the effect of microRNA‐329 (miR‐329) on nigral dopaminergic neurons in a rat model of PD via the FoxO3a signaling pathway by binding to CDKN2D. Brain tissues from the substantia nigra were taken from the rats in two groups. TUNEL staining was used to observe tyrosine hydroxylase (TH)‐positive neurons. Nigral dopaminergic neurons were randomized into the normal, blank, negative control (NC), miR‐329 mimics, miR‐329 inhibitors, small interfering (siRNA)‐CDKN2D, and miR‐329 inhibitors + siRNA‐CDKN2D groups. Expressions of miR‐329, CDKN2D, FoxO3a, AKT, caspase‐3 and Bcl‐2 were determined using RT‐qPCR and western blotting. Apoptosis rate of nigral dopaminergic neurons in 7 groups was determined by flow cytometry. Compared with the blank and NC groups, the miR‐329 mimics group showed increased miR‐329 and caspase‐3 expressions as well as decreased expressions of CDKN2D, FoxO3a, AKT, and Bcl‐2, the siRNA‐CDKN2D group indicated enhanced expressions of caspase‐3 and declined expressions of CDKN2D, FoxO3a, AKT, and Bcl‐2, and the miR‐329 inhibitors group revealed decreased miR‐329 and caspase‐3 expressions and increased expressions of CDKN2D, FoxO3a, AKT, and Bcl‐2. The apoptosis rate of nigral dopaminergic neurons was significantly increased in the miR‐329 mimics and siRNA‐CDKN2D groups, but was decreased in the miR‐329 inhibitors group. Our data suggested that downregulated miR‐329 could inhibit apoptosis of nigral dopaminergic neurons in a rat model of PD by upregulating the expression of CDKN2D via the activation of the FoxO3a signaling pathway. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8617-8629, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26608   open full text
• Unexpected regulation pattern of the IKKβ/NF‐κB/MuRF1 pathway with remarkable muscle plasticity in the Daurian ground squirrel (Spermophilus dauricus).
  Yanhong Wei, Lingchen Gong, Weiwei Fu, Shenhui Xu, Zhe Wang, Jie Zhang, Er Ning, Hui Chang, Huiping Wang, Yunfang Gao.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ As a typical hibernator, the Daurian ground squirrel (Spermophilus dauricus) spends considerable time in a state of reduced activity with prolonged fasting. Despite this, they experience little muscle atrophy and have thus become an interesting anti‐disuse muscle atrophy model. The IKKβ/NF‐κB signaling pathway is significant to muscle atrophy due to the protein degradation resulting from the upregulation of the E3 ubiquitin ligase MuRF1. The current study showed that the IKKβ/NF‐κB signaling pathway and MuRF1 maintained relatively steady mRNA and protein expression levels, with little muscle atrophy observed in the soleus (slow‐twitch, SOL) or extensor digitorum longus (fast‐twitch, EDL) during hibernation (HIB); however, mRNA expression significantly increased in the SOL and EDL muscle during interbout arousal (IBA), as did the MuRF1 mRNA level in the SOL and MuRF1 protein level in the EDL. Interestingly, the expressions of p50 and MuRF1 significantly increased during HIB in the gastrocnemius (mixed muscle, GAS) and showed moderate atrophy, but dramatically decreased during IBA. Elevated IKKβ and p50 mRNA and protein expression in the cardiac muscle (CM) during HIB did not accompany increased MuRF1 expression or muscle wasting. Importantly, almost all increased or decreased indicators in the tested tissues recovered to pre‐hibernation levels after HIB. This is the first study to report on the unexpected regulation of the IKKβ/NF‐κB/MuRF1 pathway with remarkable muscle plasticity in Daurian ground squirrels during hibernation. Furthermore, we found that different types of muscles exhibited different strategies to cope with prolonged hibernation‐induced disuse muscle atrophy. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8711-8722, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26751   open full text
• Effects of bone marrow mesenchymal stem cells transfected with Ang‐1 gene on hyperoxia‐induced optic nerve injury in neonatal mice.
  Fang‐Yu Liu, Guang‐Wu Li, Chang‐Hua Sun, Sha Chen, Jun‐Fei Cao, Qian‐Qian Ma, Sheng‐Yun Fang.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Optic nerve injury triggered retinal ganglion cell (RGC) death and optic nerve atrophy lead to visual loss. Bone marrow mesenchymal stem cells (BMSCs) are stromal cells, capable of proliferating and differentiating into different types of tissues. This aims of this study is to investigate the role of BMSCs transfected with angiopoietin‐1 (Ang‐1) in optic nerve injury induced by hyperoxia in a neonatal mice model. Ang‐1 overexpression vector was constructed and used to transfect BMSCs. Reverse transcription‐quantitative polymerase chain reaction was performed to detect Ang‐1 expression in BMSCs. The hyperoxia‐induced optic nerve injury model was established. The optic nerves at 6–7 mm posterior to the eyeball were extracted, and were treated with luxol fast blue staining, immunohistochemistry, immunofluorescence, and transmission electron microscopy to examine the effects of Ang‐1‐modified BMSCs on optic nerve injury induced by hyperoxia. The mice in the Ang‐1 + BMSCs and BMSCs groups showed remarkably improved myelin sheaths of nerve fibers compared to the hyperoxia saline group. The positive expression and integrated optic density of Ang‐1 in the Ang‐1 + BMSCs group were significantly higher compared to the air control, hyperoxia saline and BMSCs groups. The number and diameter of myelinated nerve fibers, the diameter of axons and the thickness of myelin sheath in the air control and Ang‐1 + BMSCs groups were higher compared to the hyperoxia saline group. Our study provides evidence supporting that Ang‐1‐modified BMSCs may have preventive and therapeutic effects on hyperoxia‐induced optic nerve injury in neonatal mice. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8567-8577, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26501   open full text
• Endocytosed factor V is trafficked to CD42b+ proplatelet extensions during differentiation of human umbilical cord blood‐derived megakaryocytes.
  Jacqueline M. Gertz, Kelley C. McLean, Beth A. Bouchard.
  Journal of Cellular Physiology. May 15, 2018
  --- - |2+ Plasma‐ and platelet‐derived factor Va are essential for thrombin generation catalyzed by the prothrombinase complex; however, several observations demonstrate that the platelet‐derived cofactor, which is formed following megakaryocyte endocytosis and modification of the plasma procofactor, factor V, is more hemostatically relevant. Factor V endocytosis, as a function of megakaryocyte differentiation and proplatelet formation, was assessed by flow cytometry and microscopy in CD34+ hematopoietic progenitor cells isolated from human umbilical cord blood and cultured for 12 days in the presence of cytokines to induce ex vivo differentiation into megakaryocytes. Expression of an early marker of megakaryocyte differentiation, CD41, endocytosis of factor V, and the percentage of CD41+ cells that endocytosed factor V increased from days 6 to 12 of differentiation. In contrast, statistically significant decreases in expression of the stem cell marker, CD34, and in the percentage of CD34+ cells that endocytosed factor V were observed. A statistically significant increase in the expression of CD42b, a late marker of megakaryocyte differentiation, was also observed over time, such that by Day 12, all CD42b+ cells endocytosed factor V and expressed CD41. This endocytosed factor V was trafficked to proplatelet extensions and was localized in a punctate pattern in the cytoplasm consistent with its storage in α‐granules. In conclusion, loss of CD34 and expression of CD42b define cells capable of factor V endocytosis and trafficking to proplatelet extensions during differentiation of megakaryocytes ex vivo from progenitor cells isolated from umbilical cord blood. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8691-8700, November 2018.
  

  May 15, 2018   doi: 10.1002/jcp.26749   open full text
• Curcumin alleviates ischemia reperfusion‐induced late kidney fibrosis through the APPL1/Akt signaling pathway.
  Chen Hongtao, Fan Youling, Huang Fang, Peng Huihua, Zhong Jiying, Zhou Jun.
  Journal of Cellular Physiology. May 09, 2018
  --- - |2+ As a major cause of renal failure, transient renal ischemia and reperfusion induce both acute kidney injury and late fibrosis, which are the common pathological manifestations of end‐stage renal disease. Curcumin is a biologically active polyphenolic compound found in turmeric. Increasing evidence has demonstrated that curcumin has a protective action against renal fibrosis, whereas mechanisms underlying the anti‐fibrosis role of curcumin remain poorly defined. Here, we found that APPL1, an important intracellular binding partner for AdipoR, was involved in the pathogenesis of acute injury or fibrosis and was significantly upregulated by curcumin in a mouse model of ischemia reperfusion‐induced late kidney fibrosis. Moreover, Akt signaling was the specific signaling pathway identified downstream of APPL1 in the pathogenesis of fibrosis. Our in vitro experiment demonstrated that curcumin alleviates ischemia reperfusion‐induced late kidney fibrosis via the APPL1/Akt pathway. These data are helpful for understanding the anti‐fibrosis mechanism of curcumin in the pathogenesis of AKI‐induced late fibrosis. - Journal of Cellular Physiology, Volume 233, Issue 11, Page 8588-8596, November 2018.
  

  May 09, 2018   doi: 10.1002/jcp.26536   open full text
• Generation of hematopoietic cells from mouse pluripotent stem cells in a 3D culture system of self‐assembling peptide hydrogel.
  Wei Shan, Binsheng Wang, Yulin Xu, Xia Li, Xue Li, Huafang Wang, Yu Lin, Ruxiu Tie, Qianhao Zhao, Jinyong Wang, Weiyan Zheng, Yongxian Hu, Jimin Shi, Xiaohong Yu, He Huang.
  Journal of Cellular Physiology. January 31, 2018
  --- - |2+ Abstract In vitro generation of HSCs from pluripotent stem cells (PSCs) can be regarded as novel therapeutic approaches for replacing bone marrow (BM) transplantation without immune rejection or graft versus host disease(GVHD). To date, many differentiation approaches have been evaluated in terms of directing PSCs toward different hematopoietic cell types, yet, low efficiency and no function restrict the further hematopoietic differentiation study, our research aim to develop a three dimention (3D) hematopoietic differentiation approach that serve as recapitulation of embryonic development in vitro to a degree of complexity not achievable in a two dimention (2D) culture system. We first found that mouse PSCs could be efficiently induced to hematopoietic differentiation with expression of hematopoietic makers such as c‐kit, CD41 and CD45 within self‐assembling peptide hydrogel. Colony‐forming cells assay results suggested mPSCs could differentiated into multipotential progenitor cells and 3D induction system derived hematopoietic colonies owned potential of differentiating into lymphocyte cells. In addition, in vivo animal transplantation experiment showed that mPSCs(CD45.2) could embedded into NOD/SCID mice(CD45.1) with about 3% engraftment efficiency after 3 weeks transplantation. This study demonstrated that we developed the 3D induction approach that could efficiently promoted the hematopoietic differentiation of mPSCs in vitro and obtained the multipotential progenitors that possessed the short‐term engraftment potential. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  January 31, 2018   doi: 10.1002/jcp.26509   open full text
• The Importance of miRNAs and Epigenetics in Acute Lymphoblastic Leukemia Prognosis.
  Reza Ranjbar, Ansar Karimian, Masoud aghai, Mehdi Tourani, Seyed Mostafa Mir, Javid Sabour, Farhad Jadidi, Bahman Yousefi.
  Journal of Cellular Physiology. January 31, 2018
  --- - |2+ Abstract Acute lymphoblastic leukemia (ALL), one of the most common malignant human disorders, originates in different important genetic lesions in T‐cell or B‐cell progenitors. ALL is a malignant lymphoid progenitor with peak prevalence in children (2–5 years). The rate of survival when one is suffering from ALL depends on various agents including the age of the patient, responses to anti‐leukemic therapy, and cell biology. miRNAs and epigenetics are important regulatory factors in the expression of genes. miRNAs are noncoding RNA with inhibitory effectors on specific mRNA. Patterns of DNA methylation are profoundly changed in ALL by epigenetic mechanisms. The deciphering of miRNA and the epigenetic pathogenesis in ALL could revolutionize response to the therapy and outcome, and create an enormous promise for novel approaches to reduce the toxic side‐effects of intensive leukemia. Hence, pathogenetic miRNAs and epigenetics leading to the initiation and the progression of ALL are summarized in this review. This article is protected by copyright. All rights reserved - Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-.
  

  January 31, 2018   doi: 10.1002/jcp.26510   open full text
• Neural Regulation of Bone Remodeling: Identifying Novel Neural Molecules and Pathways Between Brain and Bone.
  Shishu Huang, Zhenxia Li, Yunhui Liu, Dashuang Gao, Xinzhou Zhang, Jin Hao, Fan Yang.
  Journal of Cellular Physiology. January 27, 2018
  --- - |2+ Abstract The metabolism and homeostasis of skeletal system has historically been regarded to be associated with the endocrine system. However, such view has been expanded with the recognition of several neural pathways playing important roles in the regulation of bone metabolism via central relays. In particular, bone metabolism and homeostasis has been reported to be precisely modulated by the central neural signaling. Initiated by the finding of leptin, the axis of neural regulation on bone expands rapidly. Semaphorin‐plexin system play an important role in the crosstalk between osteoclasts and osteoblasts, a complex system has also been identified and includes neuropeptide Y and cannabinoids. These findings facilitate our understanding of the central neuropeptides and neural factors in the modulation of bone metabolism and homeostasis, and these neuronal pathways also represent an area of research scenario that identifies the novel regulation between brain and bone. These regulatory mechanisms correlate with other homeostatic networks and demonstrate a more intricate and synergetic bone biology than previously envisioned. As such, this review summarizes the current knowledge of the neural regulation of bone metabolism and homeostasis as well as its role in skeletal diseases, and discusses the emerging challenges presented in this field. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  January 27, 2018   doi: 10.1002/jcp.26502   open full text
• Inhibition of glycolytic metabolism in glioblastoma cells by Pt3glc combinated with PI3K inhibitor via SIRT3‐mediated mitochondrial and PI3K/Akt‐MAPK pathway.
  Gang Wang, Xing‐Li Fu, Jun‐Jie Wang, Rui Guan, Yan Sun, Shing‐shun Tony To.
  Journal of Cellular Physiology. January 16, 2018
  --- - |2+ ABSTRACT Glioblastoma multiforme (GBM) is the most malignant and aggressive glioma with abnormal expression of genes that mediate glycolytic metabolism and tumor cell growth. Petunidin‐3‐O‐ glucoside (Pt3glc) is a kind of anthocyanin in the red grape and derived beverages, representing the most common naturally occurring anthocyanins with a reduced incidence of cancer and heart diseases. In this study, whether Pt3glc could effectively regulate glycolysis to inhibit GBM cell was investigated by using the DBTRG‐05MG cell lines. Notably, Pt3glc displayed potent anti‐proliferative activity and significantly changed the protein levels related to both glycolytic metabolism and GBM cell survival. The expression of the pro‐apoptotic protein Bax was increased with concomitant reduction on the levels of the anti‐apoptotic protein Bcl‐2 and caspase‐3 activity. Furthermore, the levels of survival signaling proteins, such as Akt and p‐Akt (Scr473), ERK and phospho‐ERK, were significantly decreased by Pt3glc in combination with the PI3K inhibitor of LY294002. Most importantly, the levels of SIRT3 and phosphorylated p53 were also down‐regulation, indicating that Pt3glc combinated with PI3K inhibitor could induced GBM cell death may act via the SIRT3/p53‐mediated mitochondrial and PI3K/Akt‐ERK pathways. Our findings thus provide rational evidence that the combination of Pt3glc with PI3K inhibitor, which target alternative pathways in GBM cells, may be a useful adjuvant therapy in glioblastoma treatment. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  January 16, 2018   doi: 10.1002/jcp.26474   open full text
• HIV‐1 increases extracellular amyloid‐beta levels through neprilysin regulation in primary cultures of human astrocytes.
  Marta Martínez‐Bonet, M Ángeles Muñoz‐Fernández, Susana Álvarez.
  Journal of Cellular Physiology. January 11, 2018
  --- - |2+ Abstract Since the success of combined antiretroviral therapy, HIV‐1‐infected individuals are now living much longer. This increased life expectancy is accompanied by a higher prevalence of HIV‐1 associated neurocognitive disorders. Rising too is the incidence in these patients of pathological hallmarks of Alzheimer's disease such as increased deposition of amyloid beta protein (Aβ). Although neurons are major sources of Aβ in the brain, astrocytes are the most numerous glial cells, therefore, even a small level of astrocytic Aβ metabolism could make a significant contribution to brain pathology. Neprilysin (NEP) is a decisive/crucial regulator of Aβ levels. We evaluated the effects of HIV‐1 on Aβ deposition and the expression and activity of NEP in primary human astrocytes. Specifically, no differences in intracellular amyloid deposits were found between infected and control cells. However, primary cultures of infected astrocytes showed more extracellular Aβ levels compared to controls. This was accompanied by reduced expression of NEP and to a significant decrease on its activity. These results indicate that the presence of HIV‐1 in the brain could contribute to the increase of the total burden of cerebral Aβ. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  January 11, 2018   doi: 10.1002/jcp.26462   open full text
• Muscle‐derived Satellite Cells for Treating Type 1 Diabetes in Rats (Rattus norvegicus).
  Yu Ren, Hefei Wang, Si Ha, Xingsheng Zhao, Xiao Wang, Yu Lan, Xiaoling Liu.
  Journal of Cellular Physiology. January 10, 2018
  --- - |2+ Abstract Diabetes mellitus(DM) is a complicated metabolic disease, with the fundamental treatment nowadays being diet control, insulin injections, slet or pancreas transplantation, which is limited because exogenous insulin injections fail to simulate normal insulin secretion in islet beta cells successfully and islet transplantation lacks organ donors. So far, stem cells with highly self‐renewal and multi‐directional differentiation potential have become a new hope for the treatment of diabetes. In this research, rat Muscle‐derived satellite cells *MDSCs*were separated and cultivated in vitro and inducted into insulin‐producing cells with observation and identification using dithizone staining and so on. T1D rat model was built with Streptozocin(STZ), then MDSCs and induced insulin‐producing cells from MDSCs labeled by Dil were transplanted respectively. Transplantation of MDSCs (group 1) and the induced insulin‐producing cells (group 2) into streptozotocin (STZ)‐induced diabetic rats resulted in lower blood glucose, lower urine glucose, higher body weight, higher glucose tolerance and less water intake and urine output than control rats (P < 0.05). Among the male rats, the blood glucose levels of 50% of group 1 rats were <16.7 mmol/L, and 40% of group 2 rats yielded similar results (P < 0.01). Among the female rats, the blood glucose levels of 66.67% of group 1 rats were <16.7 mmol/L, and 33.33% of group 2 rats showed similar blood glucose levels (P < 0.01). Histological examination revealed that the transplanted cells reached the pancreas and repaired damaged tissues. The above research conclusions provide theoretical basis, cell selection and good prospect for the treatment of diabetes. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  January 10, 2018   doi: 10.1002/jcp.26437   open full text
• Differential mechanisms of adenosine‐ and ATPγS‐induced microvascular endothelial barrier strengthening.
  Róbert Bátori, Sanjiv Kumar, Zsuzsanna Bordán, Mary Cherian‐Shaw, Anita Kovács‐Kása, Justin A. MacDonald, David J. R. Fulton, Ferenc Erdődi, Alexander D. Verin.
  Journal of Cellular Physiology. December 22, 2017
  --- - |2+ Abstract Maintenance of the endothelial cell (EC) barrier is critical to vascular homeostasis and a loss of barrier integrity results in increased vascular permeability. While the mechanisms that govern increased EC permeability have been under intense investigation over the past several decades, the processes regulating the preservation/restoration of the EC barrier remain poorly understood. Herein we show that the extracellular purines, adenosine (Ado) and ATPγS can strengthen the barrier function of human lung microvascular EC (HLMVEC). This ability involves protein kinase A (PKA) activation and decreases in myosin light chain 20 (MLC20) phosphorylation secondary to the involvement of MLC phosphatase (MLCP). In contrast to adenosine, ATPγS‐induced PKA activation is accompanied by a modest, but significant decrease in cAMP levels supporting the existence of an unconventional cAMP‐independent pathway of PKA activation. Furthermore, ATPγS‐induced EC barrier strengthening does not involve the Rap guanine nucleotide exchange factor 3 (EPAC1) which is directly activated by cAMP, but is instead dependent upon protein kinase A‐anchor protein 2 (AKAP2) expression. We also found that AKAP2 can directly interact with the myosin phosphatase‐targeting protein MYPT1 and that depletion of AKAP2 abolished ATPγS‐induced increases in transendothelial electrical resistance (TER). Adenosine‐induced strengthening of the HLMVEC barrier required the coordinated activation of PKA and EPAC1 in a cAMP‐dependent manner. In summary, ATPγS‐induced enhancement of the EC barrier is EPAC1‐independent and is instead mediated by activation of PKA which is then guided by AKAP2, in a cAMP‐independent mechanism, to activate MLCP which dephosphorylates MLC20 resulting in reduced EC contraction and preservation. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  December 22, 2017   doi: 10.1002/jcp.26419   open full text
• Effects of shRNA‐mediated Silencing of PSMA7 on Cell Proliferation and Vascular Endothelial Growth Factor Expression via the Ubiquitin‐Proteasome Pathway in Cervical Cancer.
  Chen‐Chen Ren, Li Yang, Ling Liu, Yan‐Nan Chen, Guo‐Mei Cheng, Xiao‐An Zhang, Hui Liu.
  Journal of Cellular Physiology. December 16, 2017
  --- - |2+ ABSTRACT This study aims to evaluate the effects of PSMA7 silencing on cervical cancer (CC) cell proliferation and vascular endothelial growth factor (VEGF) expression through the ubiquitin‐proteasome (UPP) pathway. CC tissue (n = 43) and normal tissues (n = 27) were first collected from patients. Human CC cell line (SiHa) and human normal cervical epithelial cells (H8) were obtained and classified into the normal, blank, negative control (NC), PSMA7‐shRNA1 and PSMA7‐shRNA2 groups. Hybridization in situ was used to detect the expressions of wild‐type and mutant p53 proteins. Immunofluorescence assay was carried out to test the activity of 20S proteasomes. RT‐qPCR and Western blotting were both performed to determine the expressions of PSMA7, ubiquitin, P27, P53 and VEGF in sample tissues and cells. MTT assay was used analyze cell proliferation rates, and flow cytometry to analyze cell cycle and apoptotic rate. Compared with normal tissue, CC tissues had increased expression levels of PSMA7, ubiquitin, p53, VEGF as well as increased the activity of 20S proteasomes, but exhibited a decrease in p27 expression. Compared with the blank and NC groups, the PSMA7‐shRNA1 and PSMA7‐shRNA2 groups all had decreased expression levels of PSMA7, ubiquitin, p53 and VEGF as well as decreased cell proliferation, 20S proteasomes activity and cell number in the S phase, increased p27 expression, cell apoptosis and cell number in the G0/G1 phase. Our study demonstrates that PSMA7 silencing can suppress CC cell proliferation and VEGF expression in addition to promoting cell apoptosis through inhibition of the UPP signaling pathway. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  December 16, 2017   doi: 10.1002/jcp.26408   open full text
• LncRNA XIST regulates myocardial infarction by targeting miR‐130a‐3p.
  Tao Zhou, Guowei Qin, Liehong Yang, Daokang Xiang, Suining Li.
  Journal of Cellular Physiology. December 11, 2017
  --- - |2+ Abstract The study was employed to probe long non‐coding RNA X‐inactive specific transcript RNA (lncRNA XIST) expression profile and its influence on cell cycle, proliferation and apoptosis in myocardial cells. We also aimed to explore the possible meditating relationship between XIST, PDE4D and miR‐130a‐3p. Gene differential analysis was carried out using Human LncRNA Microarray V3.0. Quantitative real‐time PCR (qRT‐PCR) was used to test mRNA expressions of XIST, miR‐130a‐3p and PDE4D in normal cells and post‐myocardial infarction (MI) cells. Western blot was applied to determine the protein expression profile of PED4D. Changes in viability and cell cycle/apoptosis of post‐MI myocardial cells after silencing of XIST or PDE4D were investigated by MTT assay and flow cytometry, respectively. The targeting relationship between miR‐130a‐3p and XIST, PDE4D in myocardial cells were verified by dual luciferase reporter assay. Simulated MI environment was constructed by performing anoxic preconditioning in normal cells to probe the influence of XIST on myocardial cell apoptosis. XIST and PDE4D were overexpressed in post‐MI myocardial cells, while miR‐130a‐3p was underexpressed in post‐MI myocardial cells. High‐expressed XIST and PDE4D both promoted myocardial cell apoptosis. High‐expressed XIST also inhibited myocardial cell proliferation. XIST down‐regulated miR‐130a‐3p and PDE4D was a direct target of miR‐130a‐3p. LncRNA XIST promotes MI by targeting miR‐130a‐3p. MI induced by PDE4D can be reversed by miR‐130a‐3p. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  December 11, 2017   doi: 10.1002/jcp.26327   open full text
• Eucalyptol Protects Lungs against Bacterial Invasion through Attenuating Ciliated Cell Damage and Suppressing MUC5AC Expression.
  Na Yu, Yi‐Tian Sun, Xin‐Ming Su, Miao He, Bing Dai, Jian Kang.
  Journal of Cellular Physiology. December 07, 2017
  --- - |2+ Abstract This study was conducted to investigate whether Eucalyptol plays a role in influencing bacterial growth in cigarette smoke exposed lungs. Rats were exposed to air (control) and cigarette smoke (smoking) in presence and absence of Eucalyptol (260 mg/day). Morphological analysis of lung structures and status of airway mucous production were observed under a microscope. Pathological changes of ciliated columnar epithelium in airways were examined using transmission electron microscopy. MUC5AC protein and mRNA expression in bronchoalveolar lavage fluid (BALF) and lungs were determined. Application of Eucalyptol reduced pulmonary bullae formation and airway mucus overproduction in the smoke exposed lungs. Treatment with Eucalyptol attenuated ciliated cell damage in cigarette smoke exposed lungs. Bacterial colonies of lungs were obviously lower in the Eucalyptol treated rats than that in the smoking rats (P < 0.01). Treatment with Eucalyptol reduced the counts of bacterial colonization residing in the challenged lungs (P < 0.01). Application of Eucalyptol not only decreased MUC5AC protein expression in BALF and tobacco‐exposed lungs but also suppressed its mRNA expression in the lungs (all P < 0.05). Intervention of Eucalyptol benefits elimination of bacterial organisms from tobacco exposed lungs through attenuating ciliated cell damage and suppressing MUC5AC expression in the lungs. This article is protected by copyright. All rights reserved - 'Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-. '

  December 07, 2017   doi: 10.1002/jcp.26359   open full text
• Retraction: Wang, L., Ma, X., Yan, L., Wang, T., Wen, J. and Mi, G., LncRNA SNHG1 negatively regulates miR‐145a‐5p to enhance NUAK1 expression and promote cancer cell metastasis and invasion in nasopharyngeal carcinoma. J Cell Physiol. Accepted Author Manuscript. doi:10.1002/jcp.26340.
  
  Journal of Cellular Physiology. December 07, 2017
  --- - |2+ Abstract The above article from the Journal of Cellular Physiology, published online on 7 December 2017 in Wiley Online Library (wileyonlinelibrary.com), has been withdrawn by agreement between the authors, the journal Editor in Chief, Gary Stein, and Wiley Periodicals, Inc. The withdrawal has been agreed due to the article being submitted for publication without the consent of the authors’ instructor. - Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-.
  

  December 07, 2017   doi: 10.1002/jcp.26340   open full text
• PACAP and PAC1R are differentially expressed in motor cortex of amyotrophic lateral sclerosis patients and support survival of iPSC‐derived motor neurons.
  Gabriele Bonaventura, Rosario Iemmolo, Agata G. D'Amico, Valentina La Cognata, Erminio Costanzo, Mario Zappia, Velia D'Agata, Francesca L. Conforti, Eleonora Aronica, Sebastiano Cavallaro.
  Journal of Cellular Physiology. October 20, 2017
  Amyotrophic lateral sclerosis (ALS) is a fatal and disabling neurodegenerative disease characterized by upper and lower motor neurons depletion. In our previous work, comprehensive genomic profiling of 41 motor cortex samples enabled to discriminate controls from sporadic ALS patients, and segregated these latter into two distinct subgroups (SALS1 and SALS2), each associated with different deregulated genes. In the present study, we focused our attention on two of them, Pituitary Adenylate Cyclase‐Activating Polypeptide (PACAP) and its type 1 receptor (PAC1R), and validated the results of the transcriptome experiments by quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR), immunohistochemistry and Western blot analysis. To assess the functional role of PACAP and PAC1R in ALS, we developed an in vitro model of human induced pluripotent stem cells (iPSC)‐derived motor neurons and examined the trophic effects of exogenous PACAP following neurodegenerative stimuli. Treatment with 100 nm PACAP was able to effectively rescue iPSC‐derived motor neurons from apoptosis, as shown by cell viability assay and protein dosage of the apoptotic marker (BAX). All together, these data suggest that perturbations in the PACAP‐PAC1R pathway may be involved in ALS pathology and represent a potential drug target to enhance motor neuron viability. 1) Two distinct subgroups (SALS1 and SALS2) of sporadic Amyotrophic lateral sclerosis exist and are characterized by different deregulated genes and pathways; 2) Pituitary Adenylate Cyclase‐Activating Polypeptide (PACAP) and its type 1 receptor (PAC1R) are differentially expressed in the two subgroups; 3) PACAP rescues iPSC‐derived motor neurons from apoptosis; and 4) Perturbations in the PACAP‐PAC1R pathway may be involved in ALS pathology.

  October 20, 2017   doi: 10.1002/jcp.26182   open full text
• Functional role of mesenchymal stem cells in the treatment of chronic neurodegenerative diseases.
  Debora Lo Furno, Giuliana Mannino, Rosario Giuffrida.
  Journal of Cellular Physiology. October 20, 2017
  Mesenchymal stem cells (MSCs) can differentiate into not only cells of mesodermal lineages, but also into endodermal and ectodermal derived elements, including neurons and glial cells. For this reason, MSCs have been extensively investigated to develop cell‐based therapeutic strategies, especially in pathologies whose pharmacological treatments give poor results, if any. As in the case of irreversible neurological disorders characterized by progressive neuronal death, in which behavioral and cognitive functions of patients inexorably decline as the disease progresses. In this review, we focus on the possible functional role exerted by MSCs in the treatment of some disabling neurodegenerative disorders such as Alzheimer's Disease, Amyotrophic Lateral Sclerosis, Huntington's Disease, and Parkinson's Disease. Investigations have been mainly performed in vitro and in animal models by using MSCs generally originated from umbilical cord, bone marrow, or adipose tissue. Positive results obtained have prompted several clinical trials, the number of which is progressively increasing worldwide. To date, many of them have been primarily addressed to verify the safety of the procedures but some improvements have already been reported, fortunately. Although the exact mechanisms of MSC‐induced beneficial activities are not entirely defined, they include neurogenesis and angiogenesis stimulation, antiapoptotic, immunomodulatory, and anti‐inflammatory actions. Most effects would be exerted through their paracrine expression of neurotrophic factors and cytokines, mainly delivered at damaged regions, given the innate propensity of MSCs to home to injured sites. Hopefully, in the near future more efficacious cell‐replacement therapies will be developed to substantially restore disease‐disrupted brain circuitry. The review focuses on studies using Mesenchymal Stem Cells in therapeutic strategies for chronic neurological disorders such as Alzheimer's Disease, Amyotrophic Lateral Sclerosis, Huntington's Disease, and Parkinson's Disease. The main findings obtained by in vitro and in vivo experiments, as well as in clinical trials are outlined. Possible stem cell mediated mechanisms of action are discussed.

  October 20, 2017   doi: 10.1002/jcp.26192   open full text
• Effects of microRNA‐206 and its target gene IGF‐1 on sevoflurane‐induced activation of hippocampal astrocytes in aged rats through the PI3K/AKT/CREB signaling pathway.
  Tie‐Jun Liu, Bin Wang, Qun‐Xi Li, Xiao‐ Liu Dong, Xiao‐Liang Han, Shu‐Bo Zhang.
  Journal of Cellular Physiology. October 20, 2017
  The study aims to explore the effects of microRNA‐206 (miR‐206) targeting IGF‐1 on the activation of hippocampal astrocytes in aged rats induced by sevoflurane through the PI3K/AKT/CREB signaling pathway. Wistar rats and astrocytes were divided into the normal/blank, sham/negative control (NC), sevoflurane (sevo), miR‐206 mimics + sevo, miR‐206 inhibitors + sevo, miR‐206 NC + sevo, IGF‐1 shRNA + sevo and miR‐206 inhibitors + IGF‐1 shRNA + sevo groups. The Morris water maze test was exhibited to assess the cognitive functions. Glial fibrillary acidic protein (GFAP) expression was detected by immunofluorescence assay. Western blotting and RT‐qPCR were used to detect the expression of miR‐206, IGF‐1, PI3K, AKT, CREB, pPI3K, pAKT, pCREB, cytochrome‐c (Cyt‐c) and caspase‐3. Cell viability and apoptosis were detected by MTT assay and Annexin V/PI double staining respectively. Mitochondrial transmembrane potential (MTP) were determined by flow cytometry. The IGF‐1 shRNA + sevo group showed reduced miR‐206 expression. Compared with the normal/blank group, the sevo and miR‐206 NC + sevo groups showed decreased miR‐206 and GFAP expressions, cell viability and MTP but increased expressions of IGF‐1, PI3K, AKT, CREB, pPI3K, pAKT, pCREB, Cyt‐c and caspase‐3, as well as cell apoptosis. Similar trends were observed in the miR‐206 inhibitors + sevo group when compared with the sevo group. The study provides evidence that miR‐206 alleviates the inhibition of activation of hippocampal astrocytes in aged rats induced by sevoflurane by targeting IGT‐1 through suppressing the PI3K/AKT/CREB signaling pathway. This article is protected by copyright. All rights reserved

  October 20, 2017   doi: 10.1002/jcp.26248   open full text
• Shear Stress Upregulates Regeneration‐Related Immediate Early Genes in Liver Progenitors in 3D ECM‐like Microenvironments.
  Kenichiro Nishii, Erik Brodin, Taylor Renshaw, Rachael Weesner, Emma Moran, Shay Soker, Jessica L. Sparks.
  Journal of Cellular Physiology. October 20, 2017
  The role of fluid stresses in activating the hepatic stem/progenitor cell regenerative response is not well understood. This study hypothesized that immediate early genes (IEGs) with known links to liver regeneration will be upregulated in liver progenitor cells (LPCs) exposed to in vitro shear stresses on the order of those produced from elevated interstitial flow after partial hepatectomy. The objectives were: (1) to develop a shear flow chamber for application of fluid stress to LPCs in 3D culture; and (2) to determine the effects of fluid stress on IEG expression in LPCs. Two hours of shear stress exposure at ∼4 dyn/cm2 was applied to LPCs embedded individually or as 3D spheroids within a hyaluronic acid/collagen I hydrogel. Results were compared against static controls. Quantitative reverse transcriptase polymerase chain reaction was used to evaluate the effect of experimental treatments on gene expression. Twenty‐nine genes were analyzed, including IEGs and other genes linked to liver regeneration. Four IEGs (CFOS, IP10, MKP1, ALB) and three other regeneration‐related genes (WNT, VEGF, EpCAM) were significantly upregulated in LPCs in response to fluid mechanical stress. LPCs maintained an early to intermediate stage of differentiation in spheroid culture in the absence of the hydrogel, and addition of the gel initiated cholangiocyte differentiation programs which were abrogated by the onset of flow. Collectively the flow‐upregulated genes fit the pattern of an LPC‐mediated proliferative/regenerative response. These results suggest that fluid stresses are potentially important regulators of the LPC‐mediated regeneration response in liver. This article is protected by copyright. All rights reserved

  October 20, 2017   doi: 10.1002/jcp.26246   open full text
• Curcumin, hemostasis, thrombosis and coagulation.
  Faeze Keihanian, Amin Saeidinia, Ramin Khameneh Bagheri, Thomas P. Johnston, Amirhossein Sahebkar.
  Journal of Cellular Physiology. October 20, 2017
  Atherothrombotic cardiovascular disease is a major cause of mortality throughout the world. Platelet activation and aggregation play a central role in hemostasis and thrombosis. Herbal medicines have been traditionally used in the management of cardiovascular disease and can help in modifying its progression, particularly in hemostasis and the coagulation process, as well as altering platelet function tests and some coagulation parameters. Curcumin is a polyphenol derived from the Curcuma longa plant and has been used extensively in complementary and alternative medicine, as it is nontoxic and safe with various therapeutic properties. Modern scientific research has demonstrated its anti‐inflammatory, antioxidant, anti‐carcinogenic, antithrombotic, and cardiovascular protective effects. The present study reviewed previous studies in the literature, which support the positive activity of curcumin in hemostasis, anticoagulation, and fibrinolysis. We also presented molecular mechanisms associated with the antiplatelet and anticoagulant activities of curcumin and potential implications for the treatment of cardiovascular disease. This article is protected by copyright. All rights reserved

  October 20, 2017   doi: 10.1002/jcp.26249   open full text
• CircFUT10 reduces proliferation and facilitates differentiation of myoblasts by sponging miR‐133a.
  Hui Li, Jiameng Yang, Xuefeng Wei, Chengchuang Song, Dong Dong, Yongzhen Huang, Xianyong Lan, Martin Plath, Chuzhao Lei, Yun Ma, Xinglei Qi, Yueyu Bai, Hong Chen.
  Journal of Cellular Physiology. October 18, 2017
  Circular RNAs (circRNAs) have been identified in various tissues and cell types from human, monkey, porcine and mouse. However, knowledge on circRNAs in bovine muscle development is limited. We downloaded and analyzed the circRNAs sequencing data of bovine skeletal muscle tissue, and further characterized the role of a candidate circRNA (circFUT10) in muscle development. Quantitative real‐time PCR (qPCR) and western blot assays were used to confirm the expression of genes involved in myoblasts differentiation and proliferation. Flow cytometry was performed to assess cell cycle distribution and cell apoptosis. EdU incorporation and CCK‐8 assay were performed to demonstrate cell proliferation. We demonstrated that circFUT10 was highly (but differentially) expressed in embryonic and adult skeletal muscle tissue. circFUT10 induced bovine primary myoblasts differentiation and increased the expression of MyoD, MyoG and MyhC in mRNA and protein levels. circFUT10 increased the number of myoblasts in the G0/G1 phase of the cell cycle, and decreased the proportion of cells in the S‐phase. circFUT10 inhibited the proliferation of myoblasts and promoted them apoptosis. Via a luciferase screening assay, circFUT10 is observed to sponge to miR‐133a with three potential binding sites. Specifically, we show that circFUT10 regulated myoblasts differentiation and cell survival by directly binding to miR‐133a and inhibiting miR‐133a activity. Modulation of circFUT10 expression in muscle tissue may emerge as a potential target in breeding strategies attempting to control muscle development in cattle. This article is protected by copyright. All rights reserved

  October 18, 2017   doi: 10.1002/jcp.26230   open full text
• Regulating the transcriptomes that mediate the conversion of fibroblasts to various nervous system neural cell types.
  Niusha Khazaei, Shima Rastegar‐Pouyani, Nicholas O'Toole, Ping Wee, Abdulshakour Mohammadnia, Moein Yaqubi.
  Journal of Cellular Physiology. October 17, 2017
  Our understanding of the mechanism of cell fate transition during the direct reprogramming of fibroblasts into various central nervous system (CNS) neural cell types has been limited by the lack of a comprehensive analysis on generated cells, independently and in comparison with other CNS neural cell types. Here, we applied an integrative approach on 18 independent high throughput expression data sets to gain insight into the regulation of the transcriptome during the conversion of fibroblasts into induced neural stem cells, induced neurons, induced astrocytes, and induced oligodendrocyte progenitor cells. We found common down‐regulated genes to be mostly related to fibroblast‐specific functions, and suggest their potential as markers for screening of the silencing of the fibroblast‐specific program. For example, Tagln was significantly down‐regulated across all considered data sets. In addition, we identified specific profiles of up‐regulated genes for each CNS neural cell types, which could be potential markers for maturation and efficiency screenings. Furthermore, we identified the main TFs involved in the regulation of the gene expression program during direct reprogramming. For example, in the generation of induced neurons from fibroblasts, the Rest TF was the main regulator of this reprogramming. In summary, our computational approach for meta‐analyzing independent expression data sets provides significant details regarding the molecular mechanisms underlying the regulation of the gene expression program, and also suggests potentially useful candidate genes for screening down‐regulation of fibroblast gene expression profile, maturation, and efficiency, as well as candidate TFs for increasing the efficiency of the reprogramming process. This article is protected by copyright. All rights reserved

  October 17, 2017   doi: 10.1002/jcp.26221   open full text
• Protective effects of curcumin against aflatoxicosis: a comprehensive review.
  Mohammad Mohajeri, Behzad Behnam, Arrigo F.G. Cicero, Amirhossein Sahebkar.
  Journal of Cellular Physiology. October 16, 2017
  Aflatoxicosis is a deleterious medical condition that results from aflatoxins (AFs) or ochratoxins (OTs). Contamination with these toxins exerts detrimental effects on the liver, kidneys, reproductive organs, and also on immunological and cardiovascular systems. Aflatoxicosis is closely associated with overproduction of reactive oxygen species (ROS) as key contributors to oxidative and nitrosative stress responses, and subsequent damages to lipids, proteins, RNA, and DNA. The main target organ for AF toxicity is the liver, where DNA adducts, degranulation of endoplasmic reticulum, increased hepatic lipid peroxide, GSH depletion, mitochondrial dysfunction, and reduction of enzymatic and non‐enzymatic antioxidants are manifestations of aflatoxicosis. Curcuma longa L. (turmeric) is a medicinal plant widely utilized all over the world for culinary and phytomedical purposes. Considering the antioxidant characteristic of curcumin, the main active component of turmeric, this review is intended to critically summarize the available evidence supporting possible effectiveness of curcumin against aflatoxicosis. Curcumin can serve as a promising candidate for attenuation of the adverse consequences of aflatoxicosis, acting mainly through intrinsic antioxidant effects aroused from its structure, modulation of the immune system as reflected by interleukin‐1β and transforming growth factor‐β, and interfering with AF's biotransformation by cytochrome P450 isoenzymes CYP1A, CYP3A, CYP2A, CYP2B, and CYP2C. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26212   open full text
• Quercetin Protects Against Radiocontrast Medium Toxicity In Human Renal Proximal Tubular Cells.
  Michele Andreucci, Teresa Faga, Antonio Pisani, Raffele Serra, Domenico Russo, GIovambattista De Sarro, Ashour Michael.
  Journal of Cellular Physiology. October 16, 2017
  Radiocontrast media (RCM)‐induced acute kidney injury (CI‐AKI) is a major clinical problem whose, pathophysiology is not well understood. Direct toxic effects on renal cells, possibly mediated by reactive oxygen species, have been postulated as contributing to CI‐AKI. We investigated the effect of quercetin on human renal proximal tubular (HK‐2) cells treated with the radiocontrast medium (RCM) sodium diatrizoate. Quercetin is the most widely studied flavonoid, and the most abundant flavonol present in foods. It has been suggested to have many health benefits, including angioprotective properties and anti‐cancer effects. These beneficial effects have been attributed to its antioxidant properties and its ability to modulate cell signaling pathways. Incubation of HK‐2 cells with 100μM quercetin caused a decrease in cell viability and pre‐treatment of HK‐2 cells with 100μM quercetin followed by incubation with 75mgI/ml sodium diatrizoate for 2h caused a decrease in cell viability which was worse than in cells treated with diatrizoate alone. However, further incubation of the cells (for 22 h) after removal of the diatrizoate and quercetin caused a recovery in cell viability in those cells previously treated with quercetin+diatrizoate and quercetin alone. Analysis of signaling molecules by Western blotting showed that in RCM‐treated cells receiving initial pre‐treatment with quercetin, followed by its removal, an increase in phosphorylation of Akt (Ser473), pSTAT3 (Tyr705) and FoxO3a (Thr32) as well as an induction of Pim‐1 and decrease in PARP1 cleavage were observed. Quercetin may alleviate the longer‐term toxic effects of RCM toxicity and its possible beneficial effects should be further investigated. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26213   open full text
• TIEG and estrogen modulate SOST expression in the murine skeleton.
  Malayannan Subramaniam, Kevin S. Pitel, Elizabeth S. Bruinsma, David G. Monroe, John R. Hawse.
  Journal of Cellular Physiology. October 16, 2017
  TIEG knockout (KO) mice exhibit a female‐specific osteopenic phenotype and altered expression of TIEG in humans is associated with osteoporosis. Gene expression profiling studies identified sclerostin as one of the most highly up‐regulated transcripts in the long bones of TIEG KO mice relative to WT littermates suggesting that TIEG may regulate SOST expression. TIEG was shown to substantially suppress SOST promoter activity and the regulatory elements through which TIEG functions were identified using promoter deletion and chromatin immunoprecipitation assays. Knockdown of TIEG in IDG‐SW3 osteocyte cells using shRNA and CRISPR‐Cas9 technology resulted in increased SOST expression and delayed mineralization, mimicking the results obtained from TIEG KO mouse bones. Given that TIEG is an estrogen regulated gene, and since changes in the hormonal milieu affect SOST expression, we performed ovariectomy (OVX) and estrogen replacement therapy (ERT) studies in WT and TIEG KO mice followed by miRNA and mRNA sequencing of cortical and trabecular compartments of femurs. SOST expression levels were considerably higher in cortical bone compared to trabecular bone. In cortical bone, SOST expression was increased following OVX only in WT mice and was suppressed following ERT in both genotypes. In contrast, SOST expression in trabecular bone was decreased following OVX and significantly increased following ERT. Interestingly, a number of miRNAs that are predicted to target sclerostin exhibited inverse expression levels in response to OVX and ERT. These data implicate important roles for TIEG and estrogen‐regulated miRNAs in modulating SOST expression in bone. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26211   open full text
• Discovery and Characterization of Novel Trans‐Spliced Products of Human Polyoma JC Virus Late Transcripts from PML Patients.
  A. Sami Saribas, Julia DeVoto, Akhil Golla, Hassen S. Wollebo, Martyn K. White, Mahmut Safak.
  Journal of Cellular Physiology. October 16, 2017
  Although the human neurotropic polyomavirus, JC virus (JCV), was isolated almost a half century ago, understanding the molecular mechanisms governing its biology remains highly elusive. JCV infects oligodendrocytes and astrocytes in the central nervous system (CNS) and causes a fatal brain disease known as progressive multifocal leukoencephalopathy (PML) in immunocompromised individuals including AIDS. It has a small circular DNA genome (∼ 5 kb) and generates two primary transcripts from its early and late coding regions, producing several predicted alternatively spliced products mainly by cis‐splicing. Here, we report the discovery and characterization of two novel open reading frames (ORF1 and ORF2) associated with JCV late transcripts, generated by an unusual splicing process called trans‐splicing. These ORFs result from (i) the trans‐splicing of two different lengths of the 5'‐short coding region of VP1 between the coding regions of agnoprotein and VP2 after replacing the intron located between these two coding regions, and (ii) frame‐shifts occurring within the VP2 coding sequences terminated by a stop codon. ORF1 and ORF2 are capable of encoding 58 and 72 aa long proteins respectively and are expressed in infected cells and PML patients. Each ORF protein shares a common coding region with VP1 and has a unique coding sequence of their own. When the expression of the unique coding regions of ORFs is blocked by a stop codon insertion in the viral background, the mutant virus replicates less efficiently when compared to wild‐type, suggesting that the newly discovered ORFs play critical roles in the JCV life cycle. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26219   open full text
• Differences in definitive endoderm induction approaches using growth factors and small molecules.
  Mariia S. Bogacheva, Sofia Khan, Liisa K. Kanninen, Marjo Yliperttula, Alan W. Leung, Yan‐Ru Lou.
  Journal of Cellular Physiology. October 16, 2017
  Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. Developing human liver cell models for pharmaceutical applications is highly demanding. Due to the vast number of existing protocols to generate DE cells from hPSCs, we aimed to compare the specificity and efficiency of selected published differentiation conditions. We differentiated two hPSC lines (induced PSC and embryonic stem cell) to DE cells on Matrigel matrix using growth factors (Activin A and Wnt‐3a) and small molecules (sodium butyrate and IDE 1) in different combinations. By studying dynamic changes during six days in cell morphology and the expression of markers for pluripotency, DE, and other germ layer lineages, we found that Activin A is essential for DE differentiation, while Wnt‐3a and sodium butyrate are dispensable. Although sodium butyrate exerted rapid DE differentiation kinetics, it caused massive cell death and could not generate sufficient cells for further differentiation and applications. We further discover that IDE 1 could not induce DE as reported previously. Hereby, we compared different conditions for DE induction and found an effective six day‐protocol to obtain DE cells for the further differentiation and applications. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26214   open full text
• Comprehensive transcriptome analysis of fluid shear stress altered gene expression in renal epithelial cells.
  Steven J. Kunnen, Tareq B. Malas, Cornelis M. Semeins, Astrid D. Bakker, Dorien J.M. Peters.
  Journal of Cellular Physiology. October 16, 2017
  Renal epithelial cells are exposed to mechanical forces due to flow‐induced shear stress within the nephrons. Shear stress is altered in renal diseases caused by tubular dilation, obstruction and hyperfiltration, which occur to compensate for lost nephrons. Fundamental in regulation of shear stress are primary cilia and other mechano‐sensors, and defects in cilia formation and function have profound effects on development and physiology of kidneys and other organs. We applied RNA sequencing to get a comprehensive overview of fluid‐shear regulated genes and pathways in renal epithelial cells. Functional enrichment‐analysis revealed TGF‐β, MAPK and Wnt signaling as core signaling pathways up‐regulated by shear. Inhibitors of TGF‐β and MAPK/ERK signaling modulate a wide range of mechanosensitive genes, identifying these pathways as master regulators of shear‐induced gene expression. However, the main down‐regulated pathway, i.e. JAK/STAT, is independent of TGF‐β and MAPK/ERK. Other up‐regulated cytokine pathways include FGF, HB‐EGF, PDGF and CXC. Cellular responses to shear are modified at several levels, indicated by altered expression of genes involved in cell‐matrix, cytoskeleton and glycocalyx remodeling, as well as glycolysis and cholesterol metabolism. Cilia ablation abolished shear induced expression of a subset of genes, but genes involved in TGF‐β, MAPK and Wnt signaling were hardly affected, suggesting that other mechano‐sensors play a prominent role in the shear stress response of renal epithelial cells. Modulations in signaling due to variations in fluid shear stress are relevant for renal physiology and pathology, as suggested by elevated gene expression at pathological levels of shear stress compared to physiological shear. This article is protected by copyright. All rights reserved

  October 16, 2017   doi: 10.1002/jcp.26222   open full text
• Effect of Mono–(2–Ethylhexyl) Phthalate(MEHP) on Proliferation of and Steroid Hormone Synthesis in Rat Ovarian Granulosa Cells In Vitro.
  Na Li, Te Liu, Kun Guo, Jian Zhu, Guangyan Yu, Shuyue Wang, Lin Ye.
  Journal of Cellular Physiology. October 15, 2017
  This study aimed to examine the proliferation of and secretion by rat ovarian granulosa cells (GCs) treated with mono–(2–ethylhexyl) phthalate (MEHP). Ovarian GCs were incubated with MEHP at concentration of 0, 25, 50, 100, and 200µM for 24 hours. Cell viability was determined using the MTT Cell Proliferation Assay. Progesterone and estradiol production was evaluated by radioimmunoassay (RIA) and the expression of FSHR, PR, and ER was measured by immunocytochemistry. StAR, P450scc, 3β‐HSD, 17β‐HSD, and P450arom mRNA levels were determined by RT–PCR. MEHP markedly attenuated proliferation of GCs, increased expression of sex hormone receptors and key enzymes in progesterone production, and stimulated steroid hormone secretion.The result of these analyses demonstrates that MEHP exposure of GCs may have effects on rat ovarian functions. This article is protected by copyright. All rights reserved

  October 15, 2017   doi: 10.1002/jcp.26224   open full text
• Involvement of the Nrf2/HO‐1/CO axis and therapeutic intervention with the CO‐Releasing Molecule CORM‐A1, in a murine model of Autoimmune Hepatitis.
  Katia Mangano, Eugenio Cavalli, Santa Mammana, Maria Sofia Basile, Rosario Caltabiano, Antonio Pesce, Stefano Puleo, Atanas G. Atanasov, Gaetano Magro, Ferdinando Nicoletti, Paolo Fagone.
  Journal of Cellular Physiology. October 15, 2017
  Concanavalin A (ConA)‐induced hepatitis is an experimental model of human autoimmune hepatitis induced in rodents by i.v. injection of Con A. The disease is characterized by increase in serum levels of transaminases and massive immune infiltration of the livers. Type 1, type 2 and type 17 cytokines play a pathogenic role in the development of ConA‐induced hepatitis. To understand further the immunoregulatory mechanisms operating in the development and regulation of ConA‐induced hepatitis, we have evaluated the role of the anti‐inflammatory pathway Nrf2/HO‐1/CO (Nuclear Factor E2‐related Factor 2/Heme Oxygenase‐1/Carbon Monoxide) in this condition and determined whether the in vivo administration of CO via the CO‐releasing molecule (CORM) CORM‐A1, influences serological and histological development of Con‐A‐induced hepatitis. We have firstly evaluated in silico the genes belonging to the Nrf2/HO‐1/CO pathway that are involved in the pathogenesis of autoimmune hepatitis (AIH). The data obtained from the in silico study demonstrate that a significant number of genes modulated in the liver of ConA‐challenged mice belong to the Nrf2 pathway; on the other hand, the administration of CORM‐A1 determines an improvement in several sero‐immunological and histological parameters, and it is able to modulate genes identified by the in silico analysis. Collectively, our data indicate that the Nrf2/HO‐1/CO pathway is fundamental for the regulation of the immune responses, and that therapeutic intervention aimed at its modulation by CORM‐A1 may represent a valuable strategy to be considered for the treatment of autoimmune hepatitis in humans. This article is protected by copyright. All rights reserved

  October 15, 2017   doi: 10.1002/jcp.26223   open full text
• Lipid signaling affects primary fibroblast collective migration and anchorage in response to stiffness and microtopography.
  Michael A. Mkrtschjan, Snehal B. Gaikwad, Kevin J. Kappenman, Christopher Solís, Sagar Dommaraju, Long Le, Tejal A. Desai, Brenda Russell.
  Journal of Cellular Physiology. October 15, 2017
  Cell migration is regulated by several mechanotransduction pathways, which consist of sensing and converting mechanical microenvironmental cues to internal biochemical cellular signals, such as protein phosphorylation and lipid signaling. While there has been significant progress in understanding protein changes in the context of mechanotransduction, lipid signaling is more difficult to investigate. In this study, physical cues of stiffness (10 kPa, 100 kPa, 400 kPa, and glass), and microrod or micropost topography were manipulated in order to reprogram primary fibroblasts and assess the effects of lipid signaling on the actin cytoskeleton. In an in vitro wound closure assay, primary cardiac fibroblast migration velocity was significantly higher on soft polymeric substrata. Modulation of PIP2 availability through neomycin treatment nearly doubled migration velocity on 10 kPa substrata, with significant increases on all stiffnesses. The distance between focal adhesions and the lamellar membrane (using wortmannin treatment to increase PIP2 via PI3K inhibition) was significantly shortest compared to untreated fibroblasts grown on the same surface. PIP2 localized to the leading edge of migrating fibroblasts more prominently in neomycin‐treated cells. The membrane‐bound protein, lamellipodin, did not vary under any condition. Additionally, fifteen micron‐high micropost topography, which blocks migration, concentrates PIP2 near to the post. Actin dynamics within stress fibers, measured by fluorescence recovery after photobleaching, was not significantly different with stiffness, microtopography, nor with drug treatment. PIP2‐modulating drugs delivered from microrod structures also affected migration velocity. Thus, manipulation of the microenvironment and lipid signaling regulatory drugs might be beneficial in improving therapeutics geared toward wound healing. This article is protected by copyright. All rights reserved

  October 15, 2017   doi: 10.1002/jcp.26236   open full text
• LncRNA DGCR5 promotes lung adenocarcinoma (LUAD) progression via inhibiting hsa‐mir‐22‐3p.
  Hui‐Xing Dong, Ren Wang, Jian Zeng, Jing Pan.
  Journal of Cellular Physiology. October 14, 2017
  Long non‐coding RNAs (lncRNAs) serve critical roles in the pathogenesis of various cancers, including lung adenocarcinoma (LUAD). Herein, in this study, we aimed to investigate the biological and clinical significance of lncRNA DiGeorge syndrome critical region gene 5 (DGCR5) in LUAD. It was observed that DGCR5 was upregulated in LUAD tissues and LUAD cell lines. Inhibition of DGCR5 can prevent LUAD progression via playing anti‐apoptosis roles. Both mRNA expression and protein levels of BCL‐2 were increased by DGCR5 downregulation while reversely BAX was increased. Additionally, a novel microRNA target of DGCR5, hsa‐mir‐22‐3p was identified through bioinformatics search and confirmed by dual‐luciferase reporter system. Gain and loss‐of‐function studies were performed to verify whether DGCR5 exerts its biological functions through regulating hsa‐mir‐22‐3p in vitro. Overexpression of DGCR5 was able to reverse the tumor inhibitory effect of hsa‐mir‐22‐3p mimics. Furthermore, in vivo tests tumor xenografts were established to detect the function of DGCR5 in LADU tumorigenesis. Downregulated DGCR5 expression was greatly associated with smaller tumor size, implying a favorable prognosis of LADU patients. Taken these together, DGCR5 could be considered as a prognostic biomarker and therapeutic target in LADU diagnosis and treatment. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26215   open full text
• Genetic and epigenetic factors influencing vitamin D status.
  Afsane Bahrami, Hamid Reza Sadeghnia, Seyed‐Amir Tabatabaeizadeh, Hamidreza Bahrami‐Taghanaki, Negin Behboodi, Habibollah Esmaeili, Gordon A. Ferns, Majid Ghayour Mobarhan, Amir Avan.
  Journal of Cellular Physiology. October 14, 2017
  The global prevalence of vitamin D deficiency appears to be increasing, and the impact of this on human health is important because of the association of vitamin D insufficiency with increased risk of osteoporosis, cardiovascular disease and some cancers. There are few studies on the genetic factors that can influence vitamin D levels. In particular, the data from twin and family‐based studies have reported that circulating vitamin D concentrations are partially determined by genetic factors. Moreover, it has been shown that genetic variants (e.g., mutation) and alteration (e.g., deletion, amplification, inversion) in genes involved in the metabolism, catabolism, transport, or binding of vitamin D to it receptor, might affect vitamin D level. However, the underlying genetic determinants of plasma 25‐hydroxyvitamin D3 [25(OH)D] concentrations remain to be elucidated. Furthermore, the association between epigenetic modifications such as DNA methylation and vitamin D level has now been reported in several studies. The aim of current review was to provide an overview of the possible value of loci associated to vitamin D metabolism, catabolism, and transport as well epigenetic modification and environmental factors influencing vitamin D status. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26216   open full text
• The Small Molecule AU14022 Promotes Colorectal Cancer Cell Death via p53‐mediated G2/M‐phase Arrest and Mitochondria‐mediated Apoptosis.
  Hwani Ryu, Ky‐Youb Nam, Jae Sung Kim, Sang‐Gu Hwang, Jie‐Young Song, Jiyeon Ahn.
  Journal of Cellular Physiology. October 14, 2017
  The p53 tumor suppressor plays critical roles in cell cycle regulation and apoptotic cell death, with its activation capable of sensitizing cancer cells to radiotherapy or chemotherapy. To identify small molecules that induce apoptosis via increased p53 transcriptional activity, we used a novel in‐house library containing 96 small‐molecule compounds. Using a cell‐based screening method with a p53‐responsive luciferase‐reporter assay system involving benzoxazole derivatives, we found that AU14022 administration significantly increased p53 transcriptional activity in a concentration‐dependent manner. Treatment with AU14022 increased p53 protein expression, p53 Ser15 phosphorylation, p53‐mediated expression of downstream target genes, and apoptosis in p53‐wild‐type HCT116 human colon cancer cells, but not in p53‐knockout HCT116 cells. Additionally, p53‐wild‐type HCT116 cells treated with AU14022 exhibited mitochondrial dysfunction, including modulated expression of B‐cell lymphoma‐2 family proteins and cytochrome c release. Combination treatment with AU14022 and ionizing radiation (IR) synergistically induced apoptosis as compared with IR or AU14022 treatment alone, with further investigation demonstrating that cell cycle progression was significantly arrested at the G2/M phase following AU14022 treatment. Furthermore, in a mouse p53‐wild‐type HCT116 colon cancer xenograft model, combined treatment with AU14022 and IR inhibited tumor growth more effectively than radiation alone. Therefore, AU14022 treatment induced apoptosis through p53‐mediated cell cycle arrest involving mitochondrial dysfunction, leading to enhanced radiosensitivity in colon cancer cells. These results provide a basis for further assessments of AU14022 as a promising anticancer agent. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26234   open full text
• Estrogen regulates stemness and senescence of bone marrow stromal cells to prevent osteoporosis via ERβ‐SATB2 pathway.
  Geng Wu, Rongyao Xu, Ping Zhang, Tao Xiao, Yu Fu, Yuchao Zhang, Yifei Du, Jinhai Ye, Jie Cheng, Hongbing Jiang.
  Journal of Cellular Physiology. October 14, 2017
  Decline of pluripotency in bone marrow stromal cells (BMSCs) associated with estrogen deficiency leads to a bone formation defect in osteoporosis. Special AT‐rich sequence binding protein 2 (SATB2) is crucial for maintaining stemness and osteogenic differentiation of BMSCs. However, whether SATB2 is involved in estrogen‐deficiency associated‐osteoporosis is largely unknown. In this study, we found that estrogen mediated pluripotency and senescence of BMSCs, primarily through estrogen receptor beta (ERβ). BMSCs from the OVX rats displayed increased senescence and weaker SATB2 expression, stemness, and osteogenic differentiation, while estrogen could rescue these phenotypes. Inhibition of ERβ or ERα confirmed that SATB2 was associated with ERβ in estrogen‐mediated pluripotency and senescence of BMSCs. Furthermore, estrogen mediated the upregulation of SATB2 through the induction of ERβ binding to estrogen response elements (ERE) located at ‐488 of the SATB2 gene. SATB2 overexpression alleviated senescence and enhanced stemness and osteogenic differentiation of OVX‐BMSCs. SATB2‐modified BMSCs transplantation could prevent trabecular bone loss in an ovariectomized rat model. Collectively, our study revealed the role of SATB2 in stemness, senescence and osteogenesis of OVX‐BMSCs. Collectively, these results indicate that estrogen prevents osteoporosis by promoting stemness and osteogenesis and inhibiting senescence of BMSCs through an ERβ‐SATB2 pathway. Therefore, SATB2 is a novel anti‐osteoporosis target gene. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26233   open full text
• Transcription Factor HMG Box‐containing Protein 1 (HBP1) Modulates Mitotic Clonal Expansion (MCE) during Adipocyte Differentiation.
  Chien‐Yi Chan, Ping Yu, Feng‐Tzu Chang, Zih‐Hua Chen, Ming‐Fen Lee, Chun‐Yin Huang.
  Journal of Cellular Physiology. October 14, 2017
  Transcription factor HMG box‐containing protein 1 (HBP1) has been found to be up‐regulated in rat adipose tissue and differentiated preadipocyte; however, how HBP1 is involved in adipocyte formation remains unclear. In the present study, we demonstrated that under a standard differentiation protocol HBP1 expression fluctuates with down‐regulation in the mitotic clonal expansion (MCE) stage followed by up‐regulation in the terminal differentiation stage in both 3T3‐L1 and MEF cell models. Also, HBP1 knockdown accelerated cell cycle progression in the MCE stage, but it impaired final adipogenesis. To gain further insight into the role of HBP1 in the MCE stage, we found that the HBP1 expression pattern is reciprocal to that of C/EBPβ, and ectopic expression of HBP1suppresses C/EBPβ expression. These data indicate that HBP1 functions as a negative regulator of MCE. In contrast, when HBP1 expression was gradually elevated along with a concomitant induction of C/EBPα at the end of the MCE, HBP1 knockdown leads to a significant reduction of C/EBPα expression, suggesting that HBP1‐mediated C/EBPα expression may be needed for the termination of the cell cycle at the end of MCE for terminal differentiation. All told, our findings show that HBP1 is a key transcription factor in the already complicated regulatory cascade during adipocyte differentiation. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26237   open full text
• MiR‐145‐5p inhibits proliferation and inflammatory responses of RMC through regulating AKT/GSK pathway by targeting CXCL16.
  Junbiao Wu, Yu He, ·Yining Luo, Lei Zhang, Hua Lin, Xusheng Liu, Bihao Liu, Chunling Liang, Yuan Zhou, Jiuyao Zhou.
  Journal of Cellular Physiology. October 14, 2017
  The main pathological characteristics of chronic glomerulonephritis (CGN) are diffuse mesangial cells proliferation and inflammatory responses. Our previous studies have confirmed that miR‐145‐5p was abnormally elevated in CGN rats, but its mechanism remains unclear. Therefore, this study aimed to elucidate the mechanism of miR‐145‐5p in regulation of renal mesangial cells proliferation and inflammatory responses. In vivo study, the cationic bovine serum albumin(C‐BSA)‐induced CGN rat model was established, and the content of miR‐145‐5p in renal was examined by qRT‐PCR, meanwhile, we also determined the renal function and inflammatory infiltrate. In vitro, the cell proliferation rate, cell cycle and inflammatory changes of rat mesangial cells (RMCs) were measured. Our results suggested that miR‐145‐5p extended the G0‐G1 phase, shortened S phase, inhibited cell proliferation and suppressed inflammatory responses in RMCs. Moreover, miR‐145‐5p inhibited CXCL16 protein expression through binding the 3'‐UTR of CXCL16, suppressed AKT/GSK signaling pathway, and decreased expression of inflammation related mRNAs, such as IL‐1α, IL‐2, IL‐6, and TNF‐α mRNAs. Further, locking CXCL16 alleviated inflammatory reactions and down‐regulated AKT/GSK pathway in RMCs. Above all, we concluded that miR‐145‐5p inhibited proliferation and inflammatory responses of RMCs through regulation of AKT/GSK pathway by targeting CXCL16. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26228   open full text
• NOD1 downregulates intestinal serotonin transporter and interacts with other pattern recognition receptors.
  Elena Layunta, Eva Latorre, Raquel Forcén, Laura Grasa, Miguel Ángel Plaza, Maykel Arias, Ana I. Alcalde, José E. Mesonero.
  Journal of Cellular Physiology. October 14, 2017
  Serotonin (5‐HT) is an essential gastrointestinal modulator whose effects regulate the intestinal physiology. 5‐HT effects depend on extracellular 5‐HT bioavailability, which is controlled by the serotonin transporter (SERT) expressed in both the apical and basolateral membranes of enterocytes. SERT is a critical target for regulating 5‐HT levels and consequently, modulating the intestinal physiology. The deregulation of innate immune receptors has been extensively studied in inflammatory bowel diseases (IBD), where an exacerbated defense response to commensal microbiota is observed. Interestingly, many innate immune receptors seem to affect the serotonergic system, demonstrating a new way in which microbiota could modulate the intestinal physiology. Therefore, our aim was to analyze the effects of NOD1 activation on SERT function, as well as NOD1's interaction with other immune receptors such as TLR2 and TLR4. Our results showed that NOD1 activation inhibits SERT activity and expression in Caco‐2/TC7 cells through the extracellular signal‐regulated kinase (ERK) signaling pathway. A negative feedback between 5‐HT and NOD1 expression was also described. The results showed that TLR2 and TLR4 activation seems to regulate NOD1 expression in Caco‐2/TC7 cells. To assess the extend of cross‐talk between NOD1 and TLRs, NOD1 expression was measured in the intestinal tract (ileum and colon) of wild type mice and mice with individual knockouts of TLR2, and TLR4 as well as double knockout TLR2/TLR4 mice. Hence, we demonstrate that NOD1 acts on the serotonergic system decreasing SERT activity and molecular expression. Additionally, NOD1 expression seems to be modulated by 5‐HT and other immune receptors as TLR2 and TLR4. This study could clarify the relation between both the intestinal serotonergic system and innate immune system, and their implications in intestinal inflammation. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26229   open full text
• Annexin A1, Annexin A2, and Dyrk 1B are upregulated during GAS1‐induced cell cycle arrest.
  Pérez‐Sánchez G., Jiménez A., Quezada‐Ramírez M.A., Estudillo E., Ayala‐Sarmiento A.E., Mendoza‐Hernández G., Justino Hernández‐Soto, Hernández‐Hernández F.C., Cázares‐Raga F.E., Segovia J.
  Journal of Cellular Physiology. October 14, 2017
  GAS1 is a pleiotropic protein that has been investigated because of its ability to induce cell proliferation, cell arrest, and apoptosis, depending on the cellular or the physiological context in which it is expressed. At this point, we have information about the molecular mechanisms by which GAS1 induces proliferation and apoptosis; but very few studies have been focused on elucidating the mechanisms by which GAS1 induces cell arrest. With the aim of expanding our knowledge on this subject, we first focused our research on finding proteins that were preferentially expressed in cells arrested by serum deprivation. By using a proteomics approach and mass spectrometry analysis, we identified 17 proteins in the 2‐DE protein profile of serum deprived NIH3T3 cells. Among them, Annexin A1 (Anxa1), Annexin A2 (Anxa2), dual specificity tyrosine‐phosphorylation‐regulated kinase 1B (Dyrk1B) and Eukaryotic translation initiation factor 3, F (eIf3f) were upregulated at transcriptional level regarding proliferative NIH3T3 cells. Moreover, we demonstrated that Anxa1, Anxa2, and Dyrk1b are upregulated at both the transcriptional and translational levels by the overexpression of GAS1. Thus, our results suggest that the upregulation of Anxa1, Anxa2, and Dyrk1b could be related to the ability of GAS1 to induce cell arrest and maintain cell viability. Finally, we provided further evidence showing that GAS1 through Dyrk 1B leads not only to the arrest of NIH3T3 cells but also maintains cell viability. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26226   open full text
• Pioglitazone inhibits cancer cell growth through STAT3 inhibition and enhanced AIF expression via a PPARγ‐independent pathway.
  Masanobu Tsubaki, Tomoya Takeda, Yoshika Tomonari, Keishi Kawashima, Tatsuki Itoh, Motohiro Imano, Takao Satou, Shozo Nishida.
  Journal of Cellular Physiology. October 14, 2017
  Pioglitazone is an anti‐diabetic agent that belongs to the thiazolidinedione class, which target peroxisome proliferator‐activated receptor γ (PPARγ), a transcription factor in the nuclear receptor family. Different cancer cells expressing high levels of PPARγ and PPARγ ligands induce cell cycle arrest, cell differentiation, and apoptosis. However, the mechanisms underlying these processes remain unknown. Here, we investigated the mechanism underlying pioglitazone‐induced apoptosis in human cancer cells. We showed that at similar concentrations, pioglitazone induced death in cancer cells expressing high or low levels of PPARγ. Combined treatment of pioglitazone and GW9662, a PPARγ antagonist, did not rescue this cell death phenotype. Z‐VAD‐fmk, a pan‐caspase inhibitor, did not reverse pioglitazone‐induced apoptosis in cancer cells expressing PPARγ at high or low levels. Pioglitazone suppressed the activation of signal transducers and activator of transcription 3 (STAT3) and Survivin expression, and enhanced the apoptosis‐inducing factor (AIF) levels in these cells. Furthermore, pioglitazone enhanced the cytotoxic effect of cisplatin and oxaliplatin by suppressing Survivin and increasing AIF expression. These results indicated that pioglitazone induced apoptosis via a PPARγ‐independent pathway, thus describing pioglitazone as a potential therapeutic agent for controlling the progression of different cancers. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26225   open full text
• Synergistic effect of HIF‐1α and FoxO3a trigger cardiomyocyte apoptosis under hyperglycemic ischemia condition.
  Ya‐Fang Chen, Sudhir Pandey, Cecilia Hsuan Day, Yu‐Feng Chen, Ai‐Zhi Jiang, Tsung‐Jung Ho, Ray‐Jade Chen, Vijaya PadmaViswanadha, Wei‐Wen Kuo, Chih‐Yang Huang.
  Journal of Cellular Physiology. October 14, 2017
  Cardiomyocyte death is an important pathogenic feature of ischemia and heart failure. Through this study, we showed the synergistic role of HIF‐1α and FoxO3a in cardiomyocyte apoptosis subjected to hypoxia plus elevated glucose levels. Using gene specific small interfering RNAs (siRNA), semi‐quantitative reverse transcriptase polymerase chain reaction (RT‐PCR), western blot, immunofluorescence, nuclear and cytosolic localization and TUNEL assay techniques, we determined that combined function of HIF‐1α and FoxO3a under high glucose plus hypoxia condition lead to enhanced expression of BNIP3 inducing cardiomyocyte death. Our results highlighted the importance of the synergistic role of HIF‐1α and FoxO3a in cardiomyocyte death which may add insight into therapeutic approaches to pathophysiology associated with ischemic diabetic cardiomyopathies. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26235   open full text
• Novel nanohydrogel of hyaluronic acid loaded with quercetin alone and in combination with temozolomide as new therapeutic tool, CD44 targeted based, of glioblastoma multiforme.
  Barbarisi M, Iaffaioli RV, Armenia E, Schiavo L, De Sena G, Tafuto S, Barbarisi A, Quagliariello V.
  Journal of Cellular Physiology. October 14, 2017
  Glioblastoma multiforme is the most common and aggressive primary brain cancer with only ∼3% of patients surviving more than 3 years from diagnosis. Several mechanisms are involved in drug and radiation resistance to anticancer treatments and among them one of the most important factors is the tumour microenvironment status, characterised by cancer cell hypersecretion of interleukins and cytokines. The aim of our research was the synthesis of a nanocarrier of quercetin combined with temozolomide, to enhance the specificity and efficacy of this anticancer drug commonly used in glioblastoma treatment. The nanohydrogel increased the internalization and cytotoxicity of quercetin in human glioblastoma cells and, when co‐delivered with temozolomide, contribute to an improved anticancer effect. The nanohydrogel loaded with quercetin had the ability to recognize CD44 receptor, a brain cancer cell marker, through an energy and caveolae dependent mechanism of internalization. Moreover, nanohydrogel of quercetin was able to reduce significantly IL‐8, IL‐6 and VEGF production in pro‐inflammatory conditions with interesting implications on the mechanism of glioblastoma cells drug resistance. In summary, novel CD44 targeted polymeric based nanocarriers appear to be proficient in mediating site‐specific delivery of quercetin via CD44 receptor in glioblastoma cells. This targeted therapy lead to an improved therapeutic efficacy of temozolomide by modulating the brain tumour microenvironment. This article is protected by copyright. All rights reserved

  October 14, 2017   doi: 10.1002/jcp.26238   open full text
• Melatonin application in targeting oxidative‐induced liver injuries: a review.
  Keywan Mortezaee, Neda Khanlarkhani.
  Journal of Cellular Physiology. October 12, 2017
  It is believed that oxidative stress is a key causing factor of liver damage induced by a variety of agents, and it is a major contributing factor in almost all conditions compromising liver function, including ischemia–reperfusion injury (IRI), nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), liver fibrosis, liver cirrhosis and hepatocellular carcinoma (HCC). Liver is the organ that high concentration of melatonin (N‐acetyl‐5‐methoxytryptamine) accumulates, and it is the sole organ where circulating melatonin is metabolized. Melatonin is one of the best antioxidants that protects liver, and its metabolites also have antioxidative function. Melatonin exerts its antioxidative function directly through its radical scavenging ability and indirectly through stimulation of antioxidant enzymes. The antioxidative response from melatonin in liver affects from various factors, including its dosage, route, time and duration of administration, the type of oxidative‐induced agent and species aging. This indoleamine is also an effective and promising antioxidative choice for targeting liver IRI, NAFLD, NASH, fibrosis, cirrhosis and HCC. This article is protected by copyright. All rights reserved

  October 12, 2017   doi: 10.1002/jcp.26209   open full text
• C‐C motif chemokine ligand 23 abolishes ER stress‐ and LPS‐induced reduction in proliferation of bovine endometrial epithelial cells.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. October 12, 2017
  To reduce embryonic losses in domestic animals for economic production of livestock meat and milk, chemokines and their receptors are required for proper implantation and placentation during early pregnancy. In this study, we investigated the effects of C‐C‐motif chemokine ligand 23 (CCL23) on the proliferation of bovine endometrial (BEND) epithelial cells. CCL23 treatment improved BEND cell proliferation by enhancing PCNA and cyclin D1 expression via activation of the PI3K/AKT and MAPK signaling pathways. In addition, a combination of CCL23 and tunicamycin reversed the ER stress‐induced reduction in cell proliferation and the decreased expression of UPR‐mediated signaling proteins, including IRE1α, PERK, and ATF6α. Moreover, it regulated the lipopolysaccharide‐induced inflammation in BEND cells by inhibiting the expression of pro‐inflammatory cytokines (IL‐6 and IL‐8), and by restoring intracellular Ca2+ levels. These findings demonstrate that CCL23 improves endometrial development and uterine receptivity required for implantation and placentation during early pregnancy. This article is protected by copyright. All rights reserved

  October 12, 2017   doi: 10.1002/jcp.26210   open full text
• A New Parameter of Growth Inhibition for Cell Proliferation Assays.
  Francesco Paolo Fiorentino, Luigi Bagella, Irene Marchesi.
  Journal of Cellular Physiology. October 12, 2017
  Cell proliferation assays are performed by four decades to test the anti‐proliferative activity of natural products and synthetic compounds in cell cultures. In cancer research, they are widely employed to evaluate drug efficacy in in vitro tumor models, such as established cell lines, primary cultures and recently developed three‐dimensional tumor organoids. In this manuscript, we demonstrated that current employed parameters used by researchers to quantify in vitro growth inhibition, IC50 and GI50, lead to a misinterpretation of results based on the exponential, and not linear, proliferation of the cells in culture. Therefore, we introduce a new parameter for the analysis of growth inhibition in cell proliferation assays, termed relative population doubling capacity, that can be employed to properly quantify the anti‐proliferative activity of tested compounds and to compare drug efficacy between distinct cell models. This article is protected by copyright. All rights reserved

  October 12, 2017   doi: 10.1002/jcp.26208   open full text
• Butyrate stimulates the growth of human intestinal smooth muscle cells by activation of yes‐associated protein.
  Li‐Na Dai, Jun‐Kai Yan, Yong‐Tao Xiao, Jie Wen, Tian Zhang, Ke‐Jun Zhou, Yang Wang, Wei Cai.
  Journal of Cellular Physiology. October 11, 2017
  Intestinal smooth muscle cells play a critical role in the remodeling of intestinal structure and functional adaptation after bowel resection. Recent studies have shown that supplementation of butyrate (Bu) contributes to the compensatory expansion of a muscular layer of the residual intestine in a rodent model of short‐bowel syndrome (SBS). However, the underlying mechanism remains elusive. In this study, we found that the growth of human intestinal smooth muscle cells (HISMCs) was significantly stimulated by Bu via activation of Yes‐Associated Protein (YAP). Incubation with 0.5 mM Bu induced a distinct proliferative effect on HISMCs, as indicated by the promotion of cell cycle progression and increased DNA replication. Notably, YAP silencing by RNA interference or its specific inhibitor significantly abolished the proliferative effect of Bu on HISMCs. Furthermore, Bu induced YAP expression and enhanced the translocation of YAP from the cytoplasm to the nucleus, which led to changes in the expression of mitogenesis genes, including TEAD1, TEAD4, CTGF, and Cyr61. These results provide evidence that Bu stimulates the growth of human intestinal muscle cells by activation of YAP, which may be a potential treatment for improving intestinal adaptation. Butyrate promotes the proliferation of human intestinal smooth muscle cells via enhancing the intranuclear expression of YAP.

  October 11, 2017   doi: 10.1002/jcp.26149   open full text
• Heparan sulfate proteoglycan deficiency up‐regulates the intracellular production of nitric oxide in Chinese hamster ovary cell lines.
  Sheyla V. Lucena, Gioconda E. D. D. Moura, Tiago Rodrigues, Carolina M. Watashi, Fabiana H. Melo, Marcelo Y. Icimoto, Gustavo M. Viana, Helena B. Nader, Hugo P. Monteiro, Ivarne L. S. Tersariol, Fernando T. Ogata.
  Journal of Cellular Physiology. October 09, 2017
  We investigated the role of glycosaminoglycans (GAGs) in the regulation of endothelial nitric oxide synthase (eNOS) activity in wild‐type CHO‐K1 cells and in xylosyltransferase‐deficient CHO‐745 cells. GAGs inhibit the integrin/FAK/PI3K/AKT signaling pathway in CHO‐K1 cells, decreasing the phosphorylation of eNOS at Ser1177. Furthermore, in CHO‐K1 cells, eNOS and PKCα are localized at sphingolipid‐ and cholesterol‐rich domains in the plasma membrane called caveolae. At caveolae, PKCα activation stimulates the phosphorylation of eNOS on Thr495, resulting in further inhibition of NO production in these cells. In our data, CHO‐745 cells generate approximately 12‐fold more NO than CHO‐K1 cells. Increased NO production in CHO‐745 cells promotes higher rates of protein S‐nitrosylation and protein tyrosine nitration. Regarding reactive oxygen species (ROS) production, CHO‐745 cells show lower basal levels of superoxide (O2−) than CHO‐K1 cells. In addition, CHO‐745 cells express higher levels of GPx, Trx1, and catalase than CHO‐K1 cells, suggesting that CHO‐745 cells are in a constitutive nitrosative/oxidative stress condition. Accordingly, we showed that CHO‐745 cells are more sensitive to oxidant‐induced cell death than CHO‐K1 cells. The high concentration of NO and reactive oxygen species generated by CHO‐745 cells can induce simultaneous mitochondrial biogenesis and antioxidant gene expression. These observations led us to propose that GAGs are part of a regulatory mechanism that participates in eNOS activation and consequently regulates nitrosative/oxidative stress in CHO cells. Glycosaminoglycans deficiency increases eNOS activity and mitochondrial mass.

  October 09, 2017   doi: 10.1002/jcp.26160   open full text
• Relationship of human herpes virus 6 and multiple sclerosis: A systematic review and meta‐analysis.
  Ali Pormohammad, Taher Azimi, Fateme Falah, Ebrahim Faghihloo.
  Journal of Cellular Physiology. October 09, 2017
  Infection with human herpes viruses has been suggested to contribute to multiple sclerosis (MS), while interaction between human herpes 6 (HHV6) and MS remain unclear yet. Here, we conducted a meta‐analysis on the relationship of HHV6 infection and MS. All related studies were collected from major databases. The analyses were performed by STATA 14 and Comprehensive Meta‐Analysis V2.0 softwares. Pooled odds ratios (ORs) and 95%CIs were calculated from the raw data of the including studies by the random effects models when I2 > 50% and fix model when I2 < 50%. Thirty nine studies were included in the meta‐analysis that 34 studies used molecular assays and 7 studies used serological assays for diagnosis of HHV6 infected cases. The relationship of HHV6 and MS was significant in healthy control group by yielding a summary OR of (2.23 [1.5–3.3], p = 0.06). A significant HHV6 association with MS were in the studies with >6 score that used serum/blood sample with OR of (6.7 [95%CI 4.8–8.6], p < 0.00001) and in serological studies, IgM positive titer in other neurological diseases (OND) control group was significant with OR of (8.3 [95%CI 3–24.07], p < 0.00001). This study has been showed that there were significant relationship between MS and HHV6 infection.

  October 09, 2017   doi: 10.1002/jcp.26000   open full text
• Transition metal dependent regulation of the signal transduction cascade driving oocyte meiosis.
  Stephanie Schaefer‐Ramadan, Satanay Hubrack, Khaled Machaca.
  Journal of Cellular Physiology. October 05, 2017
  The G2‐M transition of the cell cycle requires the activation of members of the Cdc25 dual specificity phosphatase family. Using Xenopus oocyte maturation as a model system, we have previously shown that chelation of transition metals blocks meiosis progression by inhibiting Cdc25C activation. Here, using approaches that allow for the isolation of very pure and active recombinant Cdc25C, we show that Cdc25C does not bind zinc as previously reported. Additionally, we show that mutants in the disordered C‐terminal end of Cdc25C are poor initiators of meiosis, likely due to their inability to localize to the proper sub‐cellular location. We further demonstrate that the transition metal chelator, TPEN, acts on or upstream of polo‐like kinases in the oocyte to block meiosis progression. Together our results provide novel insights into Cdc25C structure‐function relationship and the role of transition metals in regulating meiosis. Transition metals are required for meiosis progression in the frog oocyte. This requirement is upstream of the activation of the dual specificity phosphatase Cdc25C, which does not bind zinc, but requires specific residues in its C‐terminus for proper function.

  October 05, 2017   doi: 10.1002/jcp.26157   open full text
• RNA binding protein Musashi‐2 regulates PIWIL1 and TBX1 in mouse spermatogenesis.
  Jessie M. Sutherland, Alexander P. Sobinoff, Barbara A. Fraser, Kate A. Redgrove, Nicole A. Siddall, Peter Koopman, Gary R. Hime, Eileen A. McLaughlin.
  Journal of Cellular Physiology. October 04, 2017
  RNA‐binding proteins (RBP) are important facilitators of post‐transcriptional gene regulation. We have previously established that nuclear overexpression of the RBP Musashi‐2 (MSI2) during male germ cell maturation is detrimental to sperm cell development and fertility. Herein we determine the genes and pathways impacted by the upregulation of Msi2. Microarray analysis and qPCR confirmed differential gene expression in factors fundamental to the cell cycle, cellular proliferation, and cell death. Similarly, comparative protein expression analysis via iTRAQ, immunoblot, and immunolocalization, identified differential expression and localization of important regulators of transcription, translation, RNA processing, and spermatogenesis. Specifically, the testis‐expressed transcription factor, Tbx1, and the piRNA regulator of gamete development, Piwil1, were both found to be targeted for translational repression by MSI2. This study provides key evidence to support a fundamental role for MSI2 in post‐transcriptional regulation during male gamete development. In this study, we have utilized comparative gene and protein expression analyses to determine the impact of Msi2 overexpression on testicular germ cell development and, identify a potential mechanism for the action of MSI2 in post‐transcriptional regulation. This detailed analysis uncovered two testis‐specific MSI2‐RNA binding targets in transcription factor Tbx1 and piRNA regulator Piwil1, with evidence for their translational repression in spermatids.

  October 04, 2017   doi: 10.1002/jcp.26168   open full text
• Comparison of genome‐wide analysis techniques to DNA methylation analysis in human cancer.
  Narges Soozangar, Mohammad R. Sadeghi, Farhad Jeddi, Mohammad H. Somi, Masoud Shirmohamadi, Nasser Samadi.
  Journal of Cellular Physiology. October 04, 2017
  DNA methylation was the first epigenetic modification to be detected in human cancers with specific relation to aberrant gene expression. Herein, DNA methylation analysis explains how epigenetic patterns affect gene expression level. Hypermethylation at tumor suppressor gene loci leads to increased tumorigenesis due to tumor suppressor gene silencing, whereas global hypomethylation of CpG islands (CGIs) is followed by genomic instability and aberrant activation of multiple oncogenes. Therefore, characterization of the genes which silenced or activated epigenetically in human tumor cells can improve our understanding of cancer biology. Different genome‐wide methodologies are applied to evaluate methylation status. Various commonly conducted techniques for this evaluation are reviewed in this paper. We provided comparative description of the procedures, advantages, and drawbacks of genome‐wide DNA methylation analysis methods and biological applications, to give information on selecting the appropriate method for different methylation studies.

  October 04, 2017   doi: 10.1002/jcp.26176   open full text
• Blunting of estrogen modulation of cardiac cellular chymase/RAS activity and function in SHR.
  Sarfaraz Ahmad, Xuming Sun, Marina Lin, Jasmina Varagic, Gisele Zapata‐Sudo, Carlos M. Ferrario, Leanne Groban, Hao Wang.
  Journal of Cellular Physiology. October 04, 2017
  The relatively low efficacy of ACE‐inhibitors in the treatment of heart failure in women after estrogen loss may be due to their inability to reach the intracellular sites at which angiotensin (Ang) II is generated and/or the existence of cell‐specific mechanisms in which ACE is not the essential processing pathway for Ang II formation. We compared the metabolic pathway for Ang II formation in freshly isolated myocytes (CMs) and non‐myocytes (NCMs) in cardiac membranes extracted from hearts of gonadal‐intact and ovariectomized (OVX) adult WKY and SHR rats. Plasma Ang II levels were higher in WKY vs. SHR (strain effect: WKY: 62 ± 6 pg/ml vs. SHR: 42 ± 9 pg/ml; p < 0.01), independent of OVX. The enzymatic activities of chymase, ACE, and ACE2 were higher in NCMs versus CMs, irrespective of whether assays were performed in cardiac membranes from WKY or SHR or in the presence or absence of OVX. E2 depletion increased chymase activity, but not ACE activity, in both CMs and NCMs. Moreover, cardiac myocyte chymase activity associated with diastolic function in WKYs and cardiac structure in SHRs while no relevant functional and structural relationships between the classic enzymatic pathway of Ang II formation by ACE or the counter‐regulatory Ang‐(1‐7) forming path from Ang II via ACE2 were apparent. The significance of these novel findings is that targeted cell‐specific chymase rather than ACE inhibition may have a greater benefit in the management of HF in women after menopause. Chymase‐mediated angiotensin II formation (or chymase activity) in cardiomyocyte cell membranes is linked to worsening of diastolic function, defined by increased Doppler‐derived filling pressures (E/e′), and increases in left ventricular (LV) mass in normotensive (WKY) and hypertensive (SHR) rats, respectively, after surgically induced menopause.

  October 04, 2017   doi: 10.1002/jcp.26179   open full text
• Inhibition of arterial medial calcification and bone mineralization by extracellular nucleotides: The same functional effect mediated by different cellular mechanisms.
  Jessal J. Patel, Dongxing Zhu, Britt Opdebeeck, Patrick D'Haese, José L. Millán, Lucie E. Bourne, Caroline P.D. Wheeler‐Jones, Timothy R. Arnett, Vicky E. MacRae, Isabel R. Orriss.
  Journal of Cellular Physiology. October 04, 2017
  Arterial medial calcification (AMC) is thought to share some outward similarities to skeletal mineralization and has been associated with the transdifferentiation of vascular smooth muscle cells (VSMCs) to an osteoblast‐like phenotype. ATP and UTP have previously been shown to inhibit bone mineralization. This investigation compared the effects of extracellular nucleotides on calcification in VSMCs with those seen in osteoblasts. ATP, UTP and the ubiquitous mineralization inhibitor, pyrophosphate (PPi), dose dependently inhibited VSMC calcification by ≤85%. Culture of VSMCs in calcifying conditions was associated with an increase in apoptosis; treatment with ATP, UTP, and PPi reduced apoptosis to levels seen in non‐calcifying cells. Extracellular nucleotides had no effect on osteoblast viability. Basal alkaline phosphatase (TNAP) activity was over 100‐fold higher in osteoblasts than VSMCs. ATP and UTP reduced osteoblast TNAP activity (≤50%) but stimulated VSMC TNAP activity (≤88%). The effects of extracellular nucleotides on VSMC calcification, cell viability and TNAP activity were unchanged by deletion or inhibition of the P2Y2 receptor. Conversely, the actions of ATP/UTP on bone mineralization and TNAP activity were attenuated in osteoblasts lacking the P2Y2 receptor. Ecto‐nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) hydrolyses ATP and UTP to produce PPi. In both VSMCs and osteoblasts, deletion of NPP1 blunted the inhibitory effects of extracellular nucleotides suggesting involvement of P2 receptor independent pathways. Our results show that although the overall functional effect of extracellular nucleotides on AMC and bone mineralization is similar there are clear differences in the cellular mechanisms mediating these actions. The extracellular nucleotides, ATP and UTP, inhibit bone mineralization and arterial medial calcification. However, the cellular mechanisms mediating these effects are different.

  October 04, 2017   doi: 10.1002/jcp.26166   open full text
• GSK‐3β inhibition suppresses instability‐induced osteolysis by a dual action on osteoblast and osteoclast differentiation.
  Mehdi Amirhosseini, Rune V. Madsen, K. Jane Escott, Mathias P. Bostrom, F. Patrick Ross, Anna Fahlgren.
  Journal of Cellular Physiology. September 28, 2017
  Currently, there are no medications available to treat aseptic loosening of orthopedic implants. Using osteoprotegerin fusion protein (OPG‐Fc), we previously blocked instability‐induced osteoclast differentiation and peri‐prosthetic osteolysis. Wnt/β‐catenin signaling, which regulates OPG secretion from osteoblasts, also modulates the bone tissue response to mechanical loading. We hypothesized that activating Wnt/β‐catenin signaling by inhibiting glycogen synthase kinase‐3β (GSK‐3β) would reduce instability‐induced bone loss through regulation of both osteoblast and osteoclast differentiation. We examined effects of GSK‐3β inhibition on regulation of RANKL and OPG in a rat model of mechanical instability‐induced peri‐implant osteolysis. The rats were treated daily with a GSK‐3β inhibitor, AR28 (20 mg/kg bw), for up to 5 days. Bone tissue and blood serum were assessed by qRT‐PCR, immunohistochemistry, and ELISA on days 3 and 5, and by micro‐CT on day 5. After 3 days of treatment with AR28, mRNA levels of β‐catenin, Runx2, Osterix, Col1α1, and ALP were increased leading to higher osteoblast numbers compared to vehicle‐treated animals. BMP‐2 and Wnt16 mRNA levels were downregulated by mechanical instability and this was rescued by GSK‐3β inhibition. Osteoclast numbers were decreased significantly after 3 days of GSK‐3β inhibition, which correlated with enhanced OPG mRNA expression. This was accompanied by decreased serum levels of TRAP5b on days 3 and 5. Treatment with AR28 upregulated osteoblast differentiation, while osteoclastogenesis was blunted, leading to increased bone mass by day 5. These data suggest that GSK‐3β inactivation suppresses osteolysis through regulating both osteoblast and osteoclast differentiation in a rat model of instability‐induced osteolysis. Inactivation of GSK‐3β enhances bone mass through both increased osteoblast differentiation and suppressed osteoclastogenesis in an animal model for mechanical instability‐induced prosthetic loosening. Decreased osteoclast differentiation induced by GSK‐3β Inhibition, correlates to an upregulation of OPG mRNA. Blocking GSK‐3β rescues instability‐induced downregulation of Wnt16 and BMP‐2 mRNA.

  September 28, 2017   doi: 10.1002/jcp.26111   open full text
• Spleen tyrosine kinase influences the early stages of multilineage differentiation of bone marrow stromal cell lines by regulating phospholipase C gamma activities.
  Joji Kusuyama, Ai Kamisono, Seong ChangHwan, Muhammad S. Amir, Kenjiro Bandow, Nahoko Eiraku, Tomokazu Ohnishi, Tetsuya Matsuguchi.
  Journal of Cellular Physiology. September 28, 2017
  Bone marrow stromal cells (BMSCs) are multipotent cells that can differentiate into adipocytes and osteoblasts. Inadequate BMSC differentiation is occasionally implicated in chronic bone metabolic disorders. However, specific signaling pathways directing BMSC differentiation have not been elucidated. Here, we explored the roles of spleen tyrosine kinase (Syk) in BMSC differentiation into adipocytes and osteoblasts. We found that Syk phosphorylation was increased in the early stage, whereas its protein expression was gradually decreased during the adipogenic and osteogenic differentiation of two mouse mesenchymal stromal cell lines, ST2 and 10T(1/2), and a human BMSC line, UE6E‐7‐16. Syk inactivation with either a pharmacological inhibitor or Syk‐specific siRNA suppressed adipogenic differentiation, characterized by decreased lipid droplet appearance and the gene expression of fatty acid protein 4 (Fabp4), peroxisome proliferator‐activated receptor γ2 (Pparg2), CCAAT/enhancer binding proteins α (C/EBPα), and C/EBPβ. In contrast, Syk inhibition promoted osteogenic differentiation, represented by increase in matrix mineralization and alkaline phosphatase (ALP) activity, as well as the expression levels of osteocalcin, runt‐related transcription factor 2 (Runx2), and distal‐less homeobox 5 (Dlx5) mRNAs. We also found that Syk‐induced signals are mediated by phospholipase C γ1 (PLCγ1) in osteogenesis and PLCγ2 in adipogenesis. Notably, Syk‐activated PLCγ2 signaling was partly modulated through B‐cell linker protein (BLNK) in adipogenic differentiation. On the other hand, growth factor receptor‐binding protein 2 (Grb2) was involved in Syk‐PLCγ1 axis in osteogenic differentiation. Taken together, these results indicate that Syk‐PLCγ signaling has a dual role in regulating the initial stage of adipogenic and osteogenic differentiation of BMSCs. Syk phosphorylation was increased in the early stage, whereas its protein expression was gradually decreased during the adipogenic and osteogenic differentiation of BMSCs. Syk‐activated PLCg2 signaling was partly modulated through BLNK in adipogenic differentiation. On the other hand, Grb2 was involved in Syk‐PLCg1 axis in osteogenic differentiation.

  September 28, 2017   doi: 10.1002/jcp.26130   open full text
• Mammalian target of rapamycin as a therapeutic target in osteoporosis.
  Gengyang Shen, Hui Ren, Ting Qiu, Zhida Zhang, Wenhua Zhao, Xiang Yu, Jinjing Huang, Jingjing Tang, De Liang, Zhensong Yao, Zhidong Yang, Xiaobing Jiang.
  Journal of Cellular Physiology. September 28, 2017
  The mechanistic target of rapamycin (mTOR) plays a key role in sensing and integrating large amounts of environmental cues to regulate organismal growth, homeostasis, and many major cellular processes. Recently, mounting evidences highlight its roles in regulating bone homeostasis, which sheds light on the pathogenesis of osteoporosis. The activation/inhibition of mTOR signaling is reported to positively/negatively regulate bone marrow mesenchymal stem cells (BMSCs)/osteoblasts‐mediated bone formation, adipogenic differentiation, osteocytes homeostasis, and osteoclasts‐mediated bone resorption, which result in the changes of bone homeostasis, thereby resulting in or protect against osteoporosis. Given the likely importance of mTOR signaling in the pathogenesis of osteoporosis, here we discuss the detailed mechanisms in mTOR machinery and its association with osteoporosis therapy. The activation/inhibition of mTOR signaling can positively/negatively regulate bone marrow mesenchymal stem cells (BMSCs)/osteoblasts‐mediated bone formation, adipogenic differentiation, osteocytes homeostasis, and osteoclasts‐mediated bone resorption, which result in the changes of bone homeostasis, thereby resulting in or protect against osteoporosis.

  September 28, 2017   doi: 10.1002/jcp.26161   open full text
• Adult‐onset brain tumors and neurodegeneration: Are polyphenols protective?
  Tiziana Squillaro, Carla Schettino, Simone Sampaolo, Umberto Galderisi, Giuseppe Di Iorio, Antonio Giordano, Mariarosa A. B. Melone.
  Journal of Cellular Physiology. September 28, 2017
  Aging is a primary risk factor for both neurodegenerative disorders (NDs) and tumors such as adult‐onset brain tumors. Since NDs and tumors are severe, disabling, progressive and often incurable conditions, they represent a pressing problem in terms of human suffering and economic costs to the healthcare systems. The current challenge for physicians and researchers is to develop new therapeutic strategies in both areas to improve the patients’ quality of life. In addition to genetics and environmental stressors, the increase in cellular oxidative stress as one of the potential common etiologies has been reported for both disorders. Recently, the scientific community has focused on the beneficial effects of dietary antioxidant classes, known as nutraceuticals, such as carotenoids, vitamins, and polyphenols. Among these compounds, polyphenols are considered to be one of the most bioactive agents in neurodegeneration and tumor prevention. Despite the beneficial activity of polyphenols, their poor bioavailability and inefficient delivery systems are the main factors limiting their use in medicine and functional food. The development of polymeric nanoparticle‐based delivery systems able to encapsulate and preserve polyphenolic compounds may represent a promising tool to enhance their stability, solubility, and cell membrane permeation. In the present review we provide an overview of the main polyphenolic compounds used for ND and brain tumor prevention and treatment that explores their mechanisms of action, recent clinical findings and principal factors limiting their application in medicine. NDs and tumors are severe, disabling, progressive, and often incurable conditions, and represent a pressing problem in terms of human suffering and economic costs to the healthcare systems. Recently, the scientific community has focused on the beneficial effects of dietary antioxidant classes, such as polyphenols, in neurodegeneration and tumor prevention. Pre‐clinical and clinical evidence shows that polyphenolic compounds are safe and able to interact in synergy with commonly used therapeutic treatment.

  September 28, 2017   doi: 10.1002/jcp.26170   open full text
• The inhibitory effect in Fraxinellone on oxidative stress‐induced senescence correlates with AMP‐activated protein kinase‐dependent autophagy restoration.
  Xiaojuan Han, Honghan Chen, Jiao Zhou, Haoran Tai, Hui Gong, Xiaobo Wang, Ning Huang, Jianqiong Qin, Tingting Fang, Fei Wang, Hengyi Xiao.
  Journal of Cellular Physiology. September 28, 2017
  As a natural metabolite of limonoids from Dictamnus dasycarpus, fraxinellone has been reported to be neuroprotective and anti‐inflammatory. However, its influence on cellular metabolism remains largely unknown. In the present study, we investigated the effect of fraxinellone on cellular senescence‐induced by oxidative stress and the potential mechanism. We found that fraxinellone administration caused growth arrest and certainly repressed the activity of senescence associated β‐galactosidase as well as the expression of senescence‐associated‐genes. Interestingly, this effect of fraxinellone is closely correlated with the restoration of impaired autophagy and the activation of AMPK. Notably, fraxinellone reacts in an AMPK‐dependent but mTORC1‐independent manner. Together, our study demonstrates for the first time that fraxinellone has the effect on senescence inhibition and AMPK activation, and supports the notion that autophagic mechanism is important for aging prevention. These findings expanded the list of natural compounds and will be potentially utilized for aging decay and/or AMPK activation. We found that fraxinellone administration caused growth arrest and certainly repressed the activity of senescence‐associated β‐galactosidase as well as the expression of senescence‐associated genes. Interestingly, this effect of fraxinellone is closely correlated with the restoration of impaired autophagy and the activation of AMPK. Notably, fraxinellone reacts in an AMPK‐dependent but mTORC1‐independent manner.

  September 28, 2017   doi: 10.1002/jcp.26169   open full text
• Neutrophil extracellular traps in acrolein promoted hepatic ischemia reperfusion injury: Therapeutic potential of NOX2 and p38MAPK inhibitors.
  Suyavaran Arumugam, Kesthuru Girish Subbiah, Kempaiah Kemparaju, Chinnasamy Thirunavukkarasu.
  Journal of Cellular Physiology. September 28, 2017
  Neutrophil is a significant contributor to ischemia reperfusion (IR) induced liver tissue damage. However, the exact role of neutrophils in IR induced innate immune activation and liver damage is not quite clear. Our study sheds light on the role of chronic oxidative stress end products in worsening the IR inflammatory process by neutrophil recruitment and activation following liver surgery. We employed specific inhibitors for molecular targets—NOX2 (NADPH oxidase 2) and P38 MAPK (Mitogen activated protein kinase) signal to counteract neutrophil activation and neutrophil extracellular trap (NET) release induced liver damage in IR injury. We found that acrolein initiated neutrophil chemotaxis and induced NET release both in vitro and in vivo. Acrolein exposure caused NET induced nuclear and mitochondrial damage in HepG2 cells as well as aggravated the IR injury in rat liver. Pretreatment with F‐apocynin and naringin, efficiently suppressed acrolein induced NET release in vitro. Notably, it suppressed the expression of inflammatory cytokines, P38MAPK‐ERK activation, and apoptotic signals in rat liver exposed to acrolein and subjected to IR. Moreover, this combination effectively attenuated acrolein induced NET release and hepatic IR injury. In the current study we have shown that the acrolein accumulation in liver due to chronic stress, is responsible for neutrophil recruitment and its activation leading to NET induced liver damage during surgery. Our study shows that therapeutic targeting of NOX2 and P38MAPK signaling in patients with chronic hepatic disorders would improve post operative hepatic function and survival. 1. Lipid peroxidation product, augment ischemia reperfusion injury in liver; 2. Neutrophil extracellular traps contribute for ischemia reperfusion injury in liver; and 3. Inhibition of NOX2 and p38MAPK suppresses acrolein aggravated hepatic ischemia reperfusion injury.

  September 28, 2017   doi: 10.1002/jcp.26167   open full text
• Overexpression of miR‐216b: Prognostic and predictive value in acute myeloid leukemia.
  Ting‐juan Zhang, De‐hong Wu, Jing‐dong Zhou, Xi‐xi Li, Wei Zhang, Hong Guo, Ji‐chun Ma, Zhao‐qun Deng, Jiang Lin, Jun Qian.
  Journal of Cellular Physiology. September 28, 2017
  Accumulating studies have shown that miR‐216b acted as a tumor suppressor and was down‐regulated in solid tumors. However, little studies revealed the role or clinical implication of miR‐216b in blood cancers. Herein, we reported miR‐216b expression and its clinical significance in patients with acute myeloid leukemia (AML). In the current study, we analyzed bone marrow (BM) miR‐216b expression in 115 de novo AML patients examined by real‐time quantitative PCR. Notably, BM miR‐216b expression was significantly up‐regulated in AML patients, and could serve as a potential biomarker distinguishing AML from controls. No significant correlations of BM miR‐216 expression were found with sex, age, white blood cells, hemoglobin, platelets, BM blasts, French–American–British classifications, and karyotypes. Significantly, patients with high miR‐216b expression tended to have a lower frequency of FLT3‐ITD mutation and higher incidence of U2AF1 and IDH1/2 mutations. Moreover, complete remission (CR) rate and overall survival were negatively affected by BM miR‐216b overexpression among cytogenetically normal AML (CN‐AML). Cox regression analyses showed that high BM miR‐216b expression may act as an independent risk factor in CN‐AML patients. Among the follow‐up patients, BM miR‐216b level in CR phase was markedly lower than in diagnosis time, and was returned in relapse phase. Collectively, our findings indicated that miR‐216b overexpression was a frequent event in de novo AML, and independently conferred a poor prognosis in CN‐AML. Moreover, miR‐216b expression was a valuable biomarker correlated with disease recurrence in AML. Collectively, our findings indicated that miR‐216b overexpression was a frequent event in de novo AML, and independently conferred a poor prognosis in CN‐AML. Moreover, miR‐216b expression was a valuable biomarker correlated with disease recurrence in AML.

  September 28, 2017   doi: 10.1002/jcp.26171   open full text
• Global analysis of gene expression profiles in the submandibular salivary gland of klotho knockout mice.
  Sung‐Min Kwon, Soo‐A Kim, Jung‐Hoon Yoon, Jong‐In Yook, Sang‐Gun Ahn.
  Journal of Cellular Physiology. September 28, 2017
  Salivary dysfunction commonly occurs in many older adults and is considered a physiological phenomenon. However, the genetic changes in salivary glands during aging have not been characterized. The present study analyzed the gene expression profile in salivary glands from accelerated aging klotho deficient mice (klotho−/−, 4 weeks old). Microarray analysis showed that 195 genes were differentially expressed (z‐score > 2 in two independent arrays) in klotho null mice compared to wild‐type mice. Importantly, alpha2‐Na+/K+‐ATPase (Atp1a2), Ca2+‐ATPase (Atp2a1), epidermal growth factor (EGF), and nerve growth factor (NGF), which have been suggested to be regulators of submandibular salivary gland function, were significantly decreased. When a network was constructed from the differentially expressed genes, proliferator‐activated receptor‐γ (PPAR γ), which regulates energy homeostasis and insulin sensitivity, was located at the core of the network. In addition, the expression of genes proposed to regulate various PPAR γ‐related cellular pathways, such as Klk1b26, Egfbp2, Cox8b, Gpx3, Fabp3, EGF, and NGFβ, was altered in the submandibular salivary glands of klotho−/− mice. Our results may provide clues for the identification of novel genes involved in salivary gland dysfunction. Further characterization of these differentially expressed genes will be useful in elucidating the genetic basis of aging‐related changes in the submandibular salivary gland. Salivary dysfunction commonly occurs in many older adults and is considered a physiological phenomenon. However, the genetic changes in salivary glands during aging have not been characterized. The present study analyzed the gene expression profile in salivary glands from accelerated aging klotho deficient mice (klotho−/−, 4 weeks old).

  September 28, 2017   doi: 10.1002/jcp.26172   open full text
• Fibroblast growth factor 2 induces proliferation and distribution of G2/M phase of bovine endometrial cells involving activation of PI3K/AKT and MAPK cell signaling and prevention of effects of ER stress.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. September 28, 2017
  Fibroblast growth factor 2 (FGF2) is abundantly expressed in conceptuses and endometria during pregnancy in diverse animal models including domestic animals. However, its intracellular mechanism of action has not been reported for bovine endometrial cells. Therefore, the aim of this study was to identify functional roles of FGF2 in bovine endometrial (BEND) cell line which has served as a good model system for investigating regulation of signal transduction following treatment with interferon‐tau (IFNT) in vitro. Results of present study demonstrated that administration of FGF2 to BEND cells increased their proliferation and regulated the cell cycle through DNA replication by an increase of PCNA and Cyclin D1. FGF2 also increased phosphorylation of AKT, P70S6K, S6, ERK1/2, JNK, and P38 in BEND cells in a dose‐dependent manner, and expression of each of those transcription factors was inhibited by their respective pharmacological inhibitor including Wormannin, U0126, and SP600125. In addition, the increase in proliferation of BEND cells and activation of the protein kinases in response to FGF2 was suppressed by BGJ398, a FGFR inhibitor. Furthermore, proliferation of BEND cells was inhibited by tunicamycin, but treatment of BEND cells with FGF2 restored proliferation of BEND cells. Consistent with this result, the stimulated unfolded protein response (UPR) regulatory proteins induced by tunicamycin were down‐regulated by FGF2. Results of this study suggest that FGF2 promotes proliferation of BEND cells and likely enhances uterine capacity and maintenance of pregnancy by activating cell signaling via the PI3K and MAPK pathways and by restoring ER stress through the FGFR. FGF2 induces proliferation of BEND cells that was confirmed by increases in PCNA in nuclei of BEND cells and a greater percent of BEND cells in G2 phase of the cell cycle via FGFR1 and activation of PI3K/AKT and MAPK signal transduction. In addition, inhibition of FGF2‐induced proliferation of BEND cells was inhibited by tunicamycin‐induced ER stressor proteins. These results indicate that FGF2 may act on uterine endometrial cells to increase proliferation and improve uterine functions during pregnancy.

  September 28, 2017   doi: 10.1002/jcp.26173   open full text
• Involvement of nutrients and nutritional mediators in mitochondrial 3‐hydroxy‐3‐methylglutaryl‐CoA synthase gene expression.
  Tania Rescigno, Anna Capasso, Mario Felice Tecce.
  Journal of Cellular Physiology. September 28, 2017
  Mitochondrial 3‐hydroxy‐3‐methylglutaryl‐CoA (HMG‐CoA) synthase (HMGCS2) catalyses the first step of ketogenesis and is critical in various metabolic conditions. Several nutrient molecules were able to differentially modulate HMGCS2 expression levels. Docosahexaenoic acid (DHA, C22:6, n‐3), eicosapentaenoic acid (EPA, C20:5, n‐3), arachidonic acid (AA, C20:4, n‐6), and glucose increased HMGCS2 mRNA and protein levels in HepG2 hepatoma cells, while fructose decreased them. The effect of n‐6 AA resulted significantly higher than that of n‐3 PUFA, but when combined all these molecules were far less efficient. Insulin reduced HMGCS2 mRNA and protein levels in HepG2 cells, even when treated with PUFA and monosaccharides. Several nuclear receptors and transcription factors are involved in HMGCS2 expression regulation. While peroxysome proliferator activated receptor α (PPAR‐α) agonist WY14643 increased HMGCS2 expression, this treatment was unable to affect PUFA‐mediated regulation of HMGCS2 expression. Forkhead box O1 (FoxO1) inhibitor AS1842856 reduced HMGCS2 expression and suppressed induction promoted by fatty acids. Cells treatment with liver X receptor alpha (LXRα) agonist T0901317 reduced HMGCS2 mRNA, indicating a role for this transcription factor as suppressor of HMGCS2 gene. Previous observations already indicated HMGCS2 expression as possible nutrition status reference: our results show that several nutrients as well as specific nutritional related hormonal conditions are able to affect significantly HMGCS2 gene expression, indicating a relevant role for PUFA, which are mostly derived from nutritional intake. These insights into mechanisms of its regulation, specifically through nutrients commonly associated with disease risk, indicate HMGCS2 expression as possible reference marker of metabolic and nutritional status. Results show that several nutrients as well as specific nutritional related hormonal conditions are able to affect significantly HMGCS2 gene expression, indicating a relevant and complex role for PUFA, which are mostly derived from nutritional intake. These insights into mechanisms of its regulation, specifically through nutrients commonly associated with disease risk, indicate HMGCS2 expression as possible reference marker of metabolic and nutritional status.

  September 28, 2017   doi: 10.1002/jcp.26177   open full text
• NAP counteracts hyperglycemia/hypoxia induced retinal pigment epithelial barrier breakdown through modulation of HIFs and VEGF expression.
  Agata G. D'Amico, Grazia Maugeri, Daniela M. Rasà, Valentina La Cognata, Salvatore Saccone, Concetta Federico, Sebastiano Cavallaro, Velia D'Agata.
  Journal of Cellular Physiology. September 28, 2017
  Diabetic macular edema (DME) is a common complication leading to a central vision loss in patients with diabetes. In this eye pathology, the hyperglycaemic/hypoxic microenvironment of pigmented epithelium is responsible for outer blood retinal barrier integrity changes. More recently, we have shown that a small peptide derived from the activity‐dependent neuroprotective protein (ADNP), known as NAP, counteracts damages occurring during progression of diabetic retinopathy by modulating HIFs/VEGF pathway. Here, we have investigated for the first time the role of this peptide on outer blood retinal barrier (BRB) integrity exposed to hyperglycaemic/hypoxic insult mimicking a model in vitro of DME. To characterize NAP role on disease's pathogenesis, we have analyzed its effect on HIFs/VEGF system in human retinal pigmented epithelial cells, ARPE‐19, grown in high glucose and low oxygen tension. The results have shown that NAP prevents outer BRB breakdown by reducing HIF1α/HIF2α, VEGF/VEGFRs, and increasing HIF3α expression, moreover it is able to reduce the percentage of apoptotic cells by modulating the expression of two death related genes, BAX and Bcl2. Further investigations are needed to determine the possible use of NAP in DME treatment. NAP counteracts damages occurring during progression of diabetic retinopathy by modulating HIFs/VEGF pathway NAP prevents outer BRB breakdown by reducing HIF1α/HIF2α and HIF3α expression.

  September 28, 2017   doi: 10.1002/jcp.25971   open full text
• Claudin7b is required for the formation and function of inner ear in zebrafish.
  Xiaohui Li, Guili Song, Yasong Zhao, Feng Zhao, Chunyan Liu, Dong Liu, Qing Li, Zongbin Cui.
  Journal of Cellular Physiology. September 27, 2017
  Zebrafish has become an excellent model for studying the development and function of inner ear. We report here a zebrafish line in which claudin 7b (cldn7b) locus is interrupted by a Tol2 transposon at its first intron. The homozygous mutants have enlarged otocysts, smaller or no otoliths, slowly formed semicircular canals, and insensitiveness to sound stimulation. These abnormal phenotypes and hearing loss of inner ear could be mostly rescued by injection of cldn7b‐mRNA into one‐cell stage homozygous mutant embryos. Mechanistically, cldn7b‐deficiency interrupted the formation of apical junction complexes (AJCs) in otic epithelial cells of inner ear and the ion‐homeostasis of endolymph, which then led to the loss of proper contact between otoliths and normally developed hair cells in utricle and saccule or aberrant mechanosensory transduction. Thus, Cldn7b is essential for the formation and proper function of inner ear through its unique role in keeping an initial integrity of otic epithelia during zebrafish embryogenesis. We report a zebrafish line in which Cldn7b gene is interrupted by a Tol2 transposon. Homozygous cldn7b‐mutants have enlarged otocysts, smaller or no otoliths, slowly formed semicircular canals, and insensitiveness to sound stimulation. Mechanistically, Cldn7b‐deficiency interrupted the formation of apical junction complexes in otic epithelial cells of inner ear and the ion‐homeostasis of endolymph, leading to the loss of proper contact between otoliths and normally developed hair cells in utricle and saccule and aberrant mechanosensory transduction.

  September 27, 2017   doi: 10.1002/jcp.26162   open full text
• Nestin expression is dynamically regulated in cardiomyocytes during embryogenesis.
  Vanessa Hertig, Adrianna Matos‐Nieves, Vidu Garg, Louis Villeneuve, Maya Mamarbachi, Laurie Caland, Angelino Calderone.
  Journal of Cellular Physiology. September 27, 2017
  The transcriptional factors implicated in the expression of the intermediate filament protein nestin in cardiomyocytes during embryogenesis remain undefined. In the heart of 9,5–10,5 day embryonic mice, nestin staining was detected in atrial and ventricular cardiomyocytes and a subpopulation co‐expressed Tbx5. At later stages of development, nestin immunoreactivity in cardiomyocytes gradually diminished and was absent in the heart of 17,5 day embryonic mice. In the heart of wild type 11,5 day embryonic mice, 54 ± 7% of the trabeculae expressed nestin and the percentage was significantly increased in the hearts of Tbx5+/− and Gata4+/− embryos. The cell cycle protein Ki67 and transcriptional coactivator Yap‐1 were still prevalent in the nucleus of nestin(+)‐cardiomyocytes identified in the heart of Tbx5+/− and Gata4+/− embryonic mice. Phorbol 12,13‐dibutyrate treatment of neonatal rat ventricular cardiomyocytes increased Yap‐1 phosphorylation and co‐administration of the p38 MAPK inhibitor SB203580 led to significant dephosphorylation. Antagonism of dephosphorylated Yap‐1 signalling with verteporfin inhibited phorbol 12,13‐dibutyrate/SB203580‐mediated nestin expression and BrdU incorporation of neonatal cardiomyocytes. Nestin depletion with an AAV9 containing a shRNA directed against the intermediate filament protein significantly reduced the number of neonatal cardiomyocytes that re‐entered the cell cycle. These findings demonstrate that Tbx5‐ and Gata4‐dependent events negatively regulate nestin expression in cardiomyocytes during embryogenesis. By contrast, dephosphorylated Yap‐1 acting via upregulation of the intermediate filament protein nestin plays a seminal role in the cell cycle re‐entry of cardiomyocytes. Based on these data, an analogous role of Yap‐1 may be prevalent in the heart of Tbx5+/− and Gata4+/− mice. In the heart of E10,5 mice, nestin staining was detected in a partially striated pattern in cardiac troponin‐T‐immunoreactive cardiomyocytes. Nestin staining was also identified in endocardial cells lining embryonic cardiomyocytes.

  September 27, 2017   doi: 10.1002/jcp.26165   open full text
• TRPC3‐mediated Ca2+ signals as a promising strategy to boost therapeutic angiogenesis in failing hearts: The role of autologous endothelial colony forming cells.
  Francesco Moccia, Angela Lucariello, Germano Guerra.
  Journal of Cellular Physiology. September 27, 2017
  Endothelial progenitor cells (EPCs) are a sub‐population of bone marrow‐derived mononuclear cells that are released in circulation to restore damaged endothelium during its physiological turnover or rescue blood perfusion after an ischemic insult. Additionally, they may be mobilized from perivascular niches located within larger arteries’ wall in response to hypoxic conditions. For this reason, EPCs have been regarded as an effective tool to promote revascularization and functional recovery of ischemic hearts, but clinical application failed to exploit the full potential of patients‐derived cells. Indeed, the frequency and biological activity of EPCs are compromised in aging individuals or in subjects suffering from severe cardiovascular risk factors. Rejuvenating the reparative phenotype of autologous EPCs through a gene transfer approach has, therefore, been put forward as an alternative approach to enhance their therapeutic potential in cardiovascular patients. An increase in intracellular Ca2+ concentration constitutes a pivotal signal for the activation of the so‐called endothelial colony forming cells (ECFCs), the only known truly endothelial EPC subset. Studies from our group showed that the Ca2+ toolkit differs between peripheral blood‐ and umbilical cord blood (UCB)‐derived ECFCs. In the present article, we first discuss how VEGF uses repetitive Ca2+ spikes to regulate angiogenesis in ECFCs and outline how VEGF‐induced intracellular Ca2+ oscillations differ between the two ECFC subtypes. We then hypothesize about the possibility to rejuvenate the biological activity of autologous ECFCs by transfecting the cell with the Ca2+‐permeable channel Transient Receptor Potential Canonical 3, which selectively drives the Ca2+ response to VEGF in UCB‐derived ECFCs. Endothelial colony forming cells (ECFCs) isolated from peripheral blood display (PB‐ECFCs) a lower pro‐angiogenic response to VEGF as compared to those deriving from umbilical cord blood (UCB‐ECFCs). The Ca2+ permeable channel TRPC3 initiates VEGF‐induced pro‐angiogenic Ca2+ oscillations in UCB‐ECFCs. We discuss the possibility to use TRPC3 to improve the reparative potential of autlogous ECFCs in cell based therapy of failing heart.

  September 27, 2017   doi: 10.1002/jcp.26152   open full text
• Multifaceted role of IL‐21 in rheumatoid arthritis: Current understanding and future perspectives.
  Palani Dinesh, Mahaboobkhan Rasool.
  Journal of Cellular Physiology. September 27, 2017
  Rheumatoid arthritis (RA) is a systemic autoimmune inflammatory disorder designated with hyperplastic synovium, bone destruction and cartilage degradation. Current therapies involve targeting major cytokines and inflammatory mediators involved in RA to alleviate the pain and provide a temporary relief. Interleukin 21 (IL‐21), a recently identified cytokine is known to possess a versatile role in modulating the cells of the RA synovium. Over the past decade, the pleiotropic role of IL‐21 in RA pathogenesis has been implicated in several aspects. T helper 17 (Th17) and follicular T helper cells (Tfh), being the key immunomodulators of the RA synovium secrete high amounts of IL‐21 during disease progression. Several studies have provided experimental evidences elucidating the multifaceted role of IL‐21 in RA disease progression. IL‐21 has the potential to activate T cells, B cells, monocytes/macrophages and synovial fibroblasts in RA pathogenesis through activation of JAK‐STAT, MAPK and PI3K/Akt signaling pathways. Till date, therapies targeting Th17 cells and its inflammatory cytokines have been under investigation and are subjected to various clinical trials. This review showcases the role of IL‐21 in RA pathogenesis and recent reports implicating its function in various immune cells, major signaling pathways, and in promoting osteoclastogenesis. IL‐21 possess the potential to activate various signaling pathways including STAT‐3, PI3K/Akt, MAPK and several other downstream elements resulting in aberrant cellular proliferation and inflammatory processes in RA pathogenesis

  September 27, 2017   doi: 10.1002/jcp.26158   open full text
• CCL18‐dependent translocation of AMAP1 is critical for epithelial to mesenchymal transition in breast cancer.
  Haiyan Li, Dawei Zhang, Jiandong Yu, Hailing Liu, Zhiping Chen, Haifeng Zhong, Yunle Wan.
  Journal of Cellular Physiology. September 27, 2017
  AMAP1 was a GTPase‐activating protein that regulates cytoskeletal structures in focal adhesions, circular dorsal ruffles, and promote cell differentiation in tumor cells. But the activation and function of AMAP1 in breast cancer remain largely unexplored. Here we show that AMAP1 was phosphorylated and translocated to plasma membrane and formed a stable complex with Pyk2 in response to CCL18. Moreover, CCL18‐dependent AMAP1 translocation interfered the AMAP1‐IKK‐β interaction, resulting in nuclear factor‐kappaB (NF‐κB) activation. Depletion of AMAP1 expression by RNAi efficiently reversed the CCL18‐induced epithelial to mesenchymal transition (EMT) of breast cancer cells and as well as CCL18‐induced adhesion, migration and invasion. Strikingly, AMAP1 overexpression was found in breast cancers that had undergone metastasis and was strongly predictive of poor prognosis in breast cancers. Given that AMAP1 mediated CCL18‐induce activation of NF‐κB and promoted breast cancer metastasis, AMAP1 may represent a therapeutic target for the eradication of breast cancer metastasis. We show that AMAP1 was phosphorylated and translocated to plasma membrane and formed a stable complex with Pyk2 in response to CCL18. CCL18‐dependent AMAP1 translocation interfered the AMAP1‐IKK‐β interaction, resulting in nuclear factor‐kappaB (NF‐κB) activation, epithelial to mesenchymal transition (EMT), adhesion, migration, and invasion. AMAP1 overexpression was found in breast cancers that had undergone metastasis and was strongly predictive of poor prognoses in breast cancers.

  September 27, 2017   doi: 10.1002/jcp.26164   open full text
• Neoadjuvant chemotherapy in triple‐negative breast cancer: A multicentric retrospective observational study in real‐life setting.
  Teresa Gamucci, Laura Pizzuti, Isabella Sperduti, Lucia Mentuccia, Angela Vaccaro, Luca Moscetti, Paolo Marchetti, Luisa Carbognin, Andrea Michelotti, Laura Iezzi, Alessandra Cassano, Antonino Grassadonia, Antonio Astone, Andrea Botticelli, Emanuela Magnolfi, Luigi Di Lauro, Domenico Sergi, Paola Fuso, Nicola Tinari, Maddalena Barba, Marcello Maugeri‐Saccà, Elisabetta Landucci, Francesca Conti, Giuseppe Sanguineti, Michele De Tursi, Gianni Iafrate, Antonio Giordano, Gennaro Ciliberto, Clara Natoli, Patrizia Vici.
  Journal of Cellular Physiology. September 27, 2017
  We aimed to assess the efficacy of neoadjuvant chemotherapy (NACT) in a cohort of 213 triple‐negative breast cancer (TNBC) patients treated in real‐world practice at eight Italian cancer centers. We computed descriptive statistics for all the variable of interest. Factors testing significant in univariate analysis were included in multivariate models. Survival data were compared by Kaplan–Meier curves and log‐rank test. The median follow‐up was 45 months. We observed 60 (28.2%) pathological complete response (pCR). The sequential anthracyclines‐taxanes‐based regimens produced the highest rate of pCR (42.6%), followed by concomitant anthracycline‐taxane (24.2%), and other regimens (15.6%) (p = 0.008). When analyzing the role of baseline Ki‐67, a 50% cut‐off was the optimal threshold value for pCR prediction (p = 0.0005). The 5‐year disease‐free survival (DFS) was 57.3% and the 5‐year overall survival (OS) was 70.8%. In patients not achieving pCR, the optimal Ki‐67 variation between biopsy and surgical specimen with prognostic relevance on long‐term outcomes was 13% (p = 0.04). Patients with a Ki‐67 reduction (rKi‐67)<13% had worse outcomes compared to those who experienced pCR or a rKi‐67≥13%. The number of NACT cycles also affected long‐term outcomes (5‐year DFS 65.7% vs 51.6% in patients having received >6 cycles compared with their counterparts, p = 0.02). In multivariate analysis, node status, grading, and bio‐pathological treatment response (including pCR and rKi‐67) impacted DFS and OS. Our results confirmed the advantage conferred by more than 6 cycles of a sequential antracycline‐taxane‐based NACT. Higher baseline Ki‐67 values shows greater predictive significance on pathogical response, while the rKi‐67 plays a prognostic role on long‐term outcomes. The sequential anthracyclines‐taxanes‐based regimens produced the highest rate of pCR (42.6%), followed by concomitant anthracycline‐taxane (24.2%), and other regimens (15.6%) (p = 0.008). Patients with a Ki‐67 reduction (rKi‐67)<13% had worse outcomes compared to those who experienced pCR or a rKi‐67≥13%. In multivariate analysis, node status, grading, and bio‐pathological treatment response (including pCR and rKi‐67) impacted DFS and OS.

  September 27, 2017   doi: 10.1002/jcp.26103   open full text
• Downregulation of P2Y12 in the superior cervical ganglia alleviates abnormal sympathetic activity after myocardial ischemia.
  Lifang Zou, Yingxin Gong, Shanhong Zhao, Zhihua Yi, Xinyao Han, Bing Wu, Bing Wu, Tianyu Jia, Lin Li, Huilong Yua, Liran Shi, Chunping Zhang, Yun Gao, Guilin Li, Hong Xu, Hui Liu, Shangdong Liang, Shuangmei Liu.
  Journal of Cellular Physiology. September 26, 2017
  Background/Aims: Superior cervical ganglia (SCG) innervate the myocardium and participate in sympathoexcitatory transmission. P2Y12 receptor is expressed in satellite glial cells (SGCs). This study seeks to clarify whether the P2Y12 receptor is involved in the sympathoexcitation reflex after myocardial ischemia (MI). Methods: MI model was induced by occlusion of the left coronary artery. P2Y12 were assayed by real time PCR and Western blotting. Results: Our results showed that expression levels of P2Y12 mRNA and protein were significantly higher in the MI group than in the sham group. Administration of P2Y12 short hairpin RNA (shRNA) caused downregulation of the P2Y12 receptor in the SCG. In MI rats plus P2Y12 shRNA treatment group, the abnormal changes in diastolic blood pressure (DBP), systolic blood pressure (SBP), heart rate (HR), electrocardiograms (ECGs), and cardiac tissue structures were alleviated. When the treatment of P2Y12 shRNA in MI rats, upregulated co‐expression values of P2Y12 and glial fibrillary acidic protein (GFAP), the upregulation of tumor necrosis factor α (TNF‐α) and phosphorylated P38 mitogen activated protein kinase (p‐P38 MAPK) in the SCG were decreased. Conclusion: Downregulation of the P2Y12 receptor in the SCG after MI may improve cardiac function by alleviating the sympathoexcitatory reflex. This article is protected by copyright. All rights reserved

  September 26, 2017   doi: 10.1002/jcp.26184   open full text
• Characterization of a novel EB1 acetylation site important for the regulation of microtubule dynamics and cargo recruitment.
  Songbo Xie, Yang Yang, Xiaochen Lin, Jun Zhou, Dengwen Li, Min Liu.
  Journal of Cellular Physiology. September 25, 2017
  Microtubule plus ends undergo highly dynamic modifications to regulate different aspects of cellular activities. Most microtubule plus‐end tracking proteins (+TIPs) are recruited to the microtubule ends by the master loading factor, end‐binding protein 1 (EB1). These proteins coordinately regulate microtubule dynamics and cellular plasticity. Acetylation is known to modulate EB1 function; however, the molecular details of EB1 acetylation remain largely unclear. We mapped the acetylation pattern of EB1 and identified several previously uncharacterized sites of EB1 acetylation. We examined the effects of lysine‐212 (K212) acetylation and found that acetylation of this site accelerates autophagy‐mediated EB1 degradation. By time‐lapse microscopy, we found that the acetylation‐deficient K212R mutant increased the percentage of fast‐growing and long‐lived microtubules. Although K212 acetylation did not affect microtubule stability in vitro and the association of EB1 with microtubules, the K212R mutant significantly promoted microtubule regrowth in cells. Coimmunoprecipitation assays further revealed that the K212 site was critical for the recruitment of different +TIP cargoes. These data thus uncover a critical role for a novel EB1 acetylation site in regulating the dynamic structure of microtubules. In this study, the authors mapped the acetylation pattern of EB1 and identified several previously uncharacterized sites of EB1 acetylation. They found that acetylation of EB1 at K212 was important for the regulation of microtubule dynamics and cargo recruitment.

  September 25, 2017   doi: 10.1002/jcp.26133   open full text
• Yes‐associated protein 1 promotes the differentiation and mineralization of cementoblast.
  Beining Yang, Hualing Sun, Fangfang Song, Yanru Wu, Jiawei Wang.
  Journal of Cellular Physiology. September 25, 2017
  Yes‐associated protein 1 (YAP1) transcriptional coactivator is a mediator of mechanosensitive signaling. Cementum, which covers the tooth root surface, continuously senses external mechanical stimulation. Cementoblasts are responsible for the mineralization and maturation of the cementum. However, the effect of YAP1 on cementoblast differentiation remains largely unknown. In this study, we initially demonstrated that YAP1 overexpression enhanced the mineralization ability of cementoblasts. YAP1 upregulated the mRNA and protein expression of several cementogenesis markers, such as alkaline phosphatase (ALP), runt‐related transcription factor 2 (Runx2), osteocalcin (OCN), and dentin matrix acidic phosphoprotein 1 (DMP1). The YAP1 overexpression group showed higher intensities of ALP and Alizarin red stain than the YAP1‐knockdown group. Unexpectedly, a sharp increase in the expression of dentin sialophosphoprotein (DSPP) was induced by the overexpression of YAP1. Knockdown of YAP1 suppressed DSPP transcriptional activity. YAP1 overexpression activated Smad‐dependent BMP signaling and slightly inhibited Erk1/2 signaling pathway activity. Treatment with specific BMP antagonist (LDN193189) prevented the upregulation of the mRNA levels of ALP, RUNX2, and OCN, as well as intensity of ALP‐stained and mineralized nodules in cementoblasts. The Erk1/2 signaling pathway inhibitor (PD 98,059) upregulated these cementogenesis markers. Thus, our study suggested that YAP1 enhanced cementoblast mineralization in vitro. YAP1 exerted its effect on the cementoblast partly by regulating the Smad‐dependent BMP and Erk1/2 signaling pathways. YAP1 enhanced cementoblast mineralization in vitro. YAP1 exerted its effect on the cementoblast partly by regulating the Smad‐dependent BMP and Erk1/2 signaling pathways.

  September 25, 2017   doi: 10.1002/jcp.26089   open full text
• Targeted therapy of human glioblastoma via delivery of a toxin through a peptide directed to cell surface nucleolin.
  Anne‐Chloé Dhez, Elisabetta Benedetti, Andrea Antonosante, Gloria Panella, Brigida Ranieri, Tiziana Marilena Florio, Loredana Cristiano, Francesco Angelucci, Francesco Giansanti, Luana Di Leandro, Michele d'Angelo, Marina Melone, Antonella De Cola, Luca Federici, Renato Galzio, Ilaria Cascone, Fabio Raineri, Annamaria Cimini, José Courty, Antonio Giordano, Rodolfo Ippoliti.
  Journal of Cellular Physiology. September 23, 2017
  Targeted anticancer therapies demand discovery of new cellular targets to be exploited for the delivery of toxic molecules and drugs. In this perspective, in the last few years, nucleolin has been identified as an interesting surface marker to be used for the therapy of glioblastoma. In this study, we investigated whether a synthetic antagonist of cell‐surface nucleolin known as N6L, previously reported to decrease both tumour growth and tumour angiogenesis in several cancer cell lines, including glioblastoma cells, as well as endothelial cells proliferation, could be exploited to deliver a protein toxin (saporin) to glioblastoma cells. The pseudopeptide N6L cross‐linked to saporin‐S6 induced internalization of the toxin inside glioblastoma cancer cells. Our results in vitro demonstrated the effectiveness of this conjugate in inducing cell death, with an ID50 four orders of magnitude lower than that observed for free N6L. Furthermore, the preliminary “in vivo” study demonstrated efficiency in reducing the tumour mass in an orthotopic mouse model of glioblastoma This article is protected by copyright. All rights reserved

  September 23, 2017   doi: 10.1002/jcp.26205   open full text
• Suppressing Angiogenesis Regulates the Irradiation‐induced Stimulation on Osteoclastogenesis in vitro.
  Ling Tong, Guoying Zhu, Jianping Wang, Ruilian Sun, Feilong He, Jianglong Zhai.
  Journal of Cellular Physiology. September 23, 2017
  Ionizing radiation‐induced bone loss is a potential health concern in radiotherapy, occupational exposure and astronauts. Although impaired bone vasculature and reduced proliferation of bone‐forming osteoblasts has been implicated in this process, it has not been clearly characterized that whether radiation affects the growth of bone‐resorbing osteoclasts. The molecular crosstalk between different cell populations in the skeletal system has not yet been elucidated in detail, especially between the increased bone resorption at early stage of post‐irradiation and bone marrow‐derived endothelial progenitor cells (BM‐EPCs). In order to further understand the mechanisms involved in radiation‐induced bone loss at the cellular level, we assessed the effects of irradiation on angiogenesis of BM‐EPCs and osteoclastogenesis of receptor activator for nuclear factor‐κB ligand (RANKL)‐stimulated RAW 264.7 cells and crosstalk between these cell populations. We herein found significantly dysfunction of BM‐EPCs in response to irradiation at a dose of 2 Gy, including inhibited proliferation, migration, tube‐forming abilities and downregulated expression of pro‐angiogenesis vascular endothelial growth factors A (VEGF A). Meanwhile, we observed that irradiation promoted osteoclastogenesis of RANKL‐stimulated RAW 264.7 cells directly or indirectly. These results provide quantitative evidences of irradiation induced osteoclastogenesis at a cellular level, and strongly suggest the involvement of osteoclastogenesis, angiogenesis and crosstalk between bone marrow cells in the radiation‐induced bone loss. This study may provide new insights for the early diagnosis and intervention of bone loss post‐irradiation.This article is protected by copyright. All rights reserved

  September 23, 2017   doi: 10.1002/jcp.26196   open full text
• Apelin/APJ system: A novel potential therapy target for kidney disease.
  Zhen Huang, Lele Wu, Linxi Chen.
  Journal of Cellular Physiology. September 21, 2017
  Apelin is an endogenous ligand of seven‐transmembrane G protein‐coupled receptor APJ. Apelin and APJ are distributed in various tissues, including the heart, lung, kidney, and even in tumor tissues. Studies show that apelin mRNA is highly expressed in the inner stripe of kidney outer medulla, which plays an important role in process of water and sodium balance. Additionally, more studies also indicate that apelin/APJ system exerts a broad range of activities in kidney. Therefore, we review the role of apelin/APJ system in kidney diseases such as renal fibrosis, renal ischemia/reperfusion injury, diabetic nephropathy, polycystic kidney disease, and hemodialysis (HD). Apelin/APJ system can improve renal interstitial fibrosis by reducing the deposition of extracellular matrix. Apelin/APJ system significantly reduces renal ischemia/reperfusion injury by inhibiting renal cell death. Apelin/APJ system involves the progression of diabetic nephropathy (DN). Apelin/APJ system also predicts the process of polycystic kidney disease. Besides, apelin/APJ system prevents some dialysis complications in HD patients. And apelin/APJ system alleviates chronic kidney disease (CKD) by inhibiting vascular calcification (VC). Overall, apelin/APJ system plays diversified roles in kidney disease and may be a potential target for the treatment of kidney disease. Studies show that apelin mRNA is highly expressed in the inner stripe of kidney outer medulla, which plays an important role in process of water and sodium balance. Additionally, more studies also indicate that apelin/APJ system may exerts a broad range of activities in kidney. Therefore, we review the role of apelin/APJ system in kidney diseases such as renal fibrosis, renal ischemia/reperfusion injury, diabetic nephropathy, polycystic kidney disease, and hemodialysis.

  September 21, 2017   doi: 10.1002/jcp.26144   open full text
• Substrate stiffness regulated migration and angiogenesis potential of A549 cells and HUVECs.
  Dan Zhao, Changyue Xue, Qianshun Li, Mengting Liu, Wenjuan Ma, Tengfei Zhou, Yunfeng Lin.
  Journal of Cellular Physiology. September 21, 2017
  Tumor tissue tends to stiffen during solid tumor progression. Substrate stiffness is known to alter cell behaviors, such as proliferation and migration, during which angiogenesis is requisite. Mono‐culture and co‐culture systems of lung cancer cell line A549 and human umbilical vein endothelial cells (HUVECs), on polydimethylsiloxane substrates (PDMS) with varying stiffness, were used for investigating the effects of substrate stiffness on the migration and angiogenesis of lung cancer. The expressions of matrix metalloproteinases (MMPs) and angiogenesis‐related growth factors were up‐regulated with the increase of substrate stiffness, whereas that of tissue inhibitor of matrix metalloproteinase (TIMPs) were down‐regulated with increasing substrate stiffness. Our data not only suggested that stiff substrate may promote the migration and angiogenesis capacities of lung cancer, but also suggested that therapeutically targeting lung tumor stiffness or response of ECs to lung tumor stiffness may help reduce migration and angiogenesis of lung tumor. This article is protected by copyright. All rights reserved

  September 21, 2017   doi: 10.1002/jcp.26189   open full text
• Involvement of sperm acetylated histones and the nuclear isoform of Glutathione peroxidase 4 in fertilization.
  Simona Pipolo, Rossella Puglisi, Valentina Mularoni, Valentina Esposito, Andrea Fuso, Marco Lucarelli, Maria T. Fiorenza, Franco Mangia, Carla Boitani.
  Journal of Cellular Physiology. September 20, 2017
  We previously demonstrated that the nuclear form of Glutathione peroxidase 4 (nGPx4) has a peculiar distribution in sperm head, being localized to nuclear matrix and acrosome and that sperm lacking nGPx4 are more prone to decondensation in vitro. In this study we have hypothesized that sperm retained acetylated histones and nGPx4 are implicated in paternal chromatin decondensation and male pronucleus formation at fertilization. Indeed, significant higher amounts of acetylated histone H4 and acetylated histone H3 were observed by both immunofluorescence and western blotting in nGPx4‐KO sperm vs WT ones. In vitro fertilization of zona pellucida‐deprived oocytes by WT sperm in the presence of trichostatin (TSA) also demonstrated that paternal histone acetylation was inversely related to the timing of sperm nucleus decondensation at fertilization. In contrast, TSA had no effect on nGPx4‐KO sperm, indicating they had a maximal level of histone acetylation. Moreover the paternally imprinted gene Igf2/H19 was hypomethylated in KO sperm compared to WT ones. The lack of nGPx4 negatively affected male fertility, causing a marked decrease in total pups and pregnancies with delivery, a significant reduction in pronuclei (PN) embryos in in vitro fertilization assays and an approximately 2 h delay in egg fertilization in vivo. Because the zona pellucida binding and fusion to oolemma of nGPx4‐KO and WT sperm were similar, the subfertility of nGPx4 sperm reflected a decreased sperm progression through egg cumulus/zona pellucida, pinpointing a defective acrosome in line with acrosomal nGPx4 localization. We conclude that paternal acetylated histones and acrosomal nGPx4 are directly involved in fertilization. Paternal histone acetylation is inversely related to the timing of sperm nucleus decondensation at fertilization ‐the lack of the nuclear isoform of Glutathione peroxidase 4 (nGPx4) localized to the acrosomal matrix negatively affects male fertility.

  September 20, 2017   doi: 10.1002/jcp.26146   open full text
• Pmepa1 induced by RANKL‐p38 MAPK pathway has a novel role in osteoclastogenesis.
  Noboru Funakubo, Xianghe Xu, Toshio Kukita, Seiji Nakamura, Hiroshi Miyamoto, Akiko Kukita.
  Journal of Cellular Physiology. September 20, 2017
  Osteoclasts are multinucleated cells formed by fusion of preosteoclasts (POCs) derived from cells of the monocyte/macrophage lineage. We have reported a culture system that supports the formation of POCs from stroma‐depleted rat bone marrow cells. Global gene expression analysis of this culture system identified genes highly expressed in POCs. Here, we have analyzed the expression and function of one of these highly expressed genes, prostate transmembrane protein androgen induced 1 (Pmepa1), a target of TGF‐β and binds Nedd4 ubiquitin ligase, which plays a role in intracellular trafficking. We show here that the expression of Pmepa1 was strongly induced by RANKL in mouse bone marrow macrophage and in the osteoclast precursor cell line RAW‐D. The expression of Pmepa1 was increased at 24 hr of culture, but was decreased at 72 hr. Pmepa1 protein was localized to intracellular vesicle membrnane of mononuclear cells, some of which were cathepsin‐K positive. RANKL‐induced expression of Pmepa1 was significantly reduced by inhibitors of p38 MAPK signaling. Pmepa1 siRNA suppressed the formation of osteoclasts in RAW‐D cells, and inhibited the expression of cathepsin K and c‐fos but not RANK. In addition, inhibition of Pmepa1 expression reduced the surface expression of RANK in RAW‐D cells induced by RANKL. These results demonstrate that Pmepa1 is induced by RANK‐p38 MAPK pathway signaling, and upregulates cell surface expression of RANK, suggesting that Pmepa1 plays a role in osteoclastogenesis and osteoclast signaling. Pmepa1, a target of TGF‐beta was identified as a highly expressed gene in rat preosteoclasts. Pmepa1 is induced by RANK‐p38 MAPK pathway signaling, and controls osteoclastogenesis by upregulating c‐fos and promoting the transport RANK to the cell surface.

  September 20, 2017   doi: 10.1002/jcp.26147   open full text
• Horizontal transfer of miR‐23a from hypoxic tumor cell colonies can induce angiogenesis.
  T.V. Sruthi, Lincy Edatt, Grace R. Raji, Haritha K., S. Sharath Shankar, Vandana Shankar, Vishnu R., Aswini P., V.B. Sameer Kumar.
  Journal of Cellular Physiology. September 20, 2017
  Neo vessel formation by angiogenesis is an important event during many pathological conditions including cancer, where it is indispensable for tumor growth and survival. Although, various pro‐angiogenic cytokines and soluble factors, secreted by tumor cells, have been reported to promote angiogenesis, recent studies have shown regulatory role of exosomes, secreted by tumor cells in the process of angiogenesis. These exosomes are capable of carrying nucleic acids, proteins etc. as their cargo. Under the light of these facts and considering the presence of miRNAs, the non‐coding RNAs capable of regulating target gene expression, as one of the major cargos in the exosomes, we investigated, whether exosomes derived from normoxic and hypoxic tumor cell colonies exhibit difference in levels of miR‐23∼27∼24 cluster members and if so, to check the significance of their horizontall transfer on the process of angiogenesis. Results of our study showed that exosomes secreted by hypoxic tumor cell colonies possess significantly higher levels of miR23a and can induce angiogenesis.. Further we have shown that exosomes secreted by cells that ectopically over express miR23a is capable of inducing angiogenesis in different angiogenic model systems such as CAM, in ovo Xenograft and HUVEC models systems. Further, mechanistic analysis revealed that miR23a driven regulation of angiogenesis is brought about by down regulation of SIRT1 in the recipient cells. Collectively, the results presented here suggest that exosomal transfer of miR23a from tumor cell colonies can induce the process of angiogenesis by targeting SIRT1 in the recipient endothelial cells. This article is protected by copyright. All rights reserved

  September 20, 2017   doi: 10.1002/jcp.26202   open full text
• Conditionally targeted deletion of PSEN1 leads to diastolic heart dysfunction.
  Xiao‐Wei Song, Qing‐Ning Yuan, Ying Tang, Mi Cao, Ya‐Feng Shen, Zhen‐Yu Zeng, Chang‐Hai Lei, SongHua Li, Xian‐Xian Zhao, Yong‐Ji Yang.
  Journal of Cellular Physiology. September 20, 2017
  Recently, PSEN1 has been reported to have mutations in dilated cardiomyopathy pedigrees. However, the function and mechanism of PSEN1 in cardiomyopathy remains unresolved. Here, we established four types of genetically modified mice to determine the function of PSEN1 in cardiac development and pathology. PSEN1 null mutation resulted in perinatal death, retardation of heart growth, ventricular dilatation, septum defects, and valvular thickening. PSEN1 knockout in adults led to decreased muscle fibers, widened sarcomere Z lines and reduced lengths of sarcomeres in cardiomyocytes. Cardiovascular loss of function of PSEN1 induced by Sm22a‐Cre or Myh6‐Cre/ER/tamoxifen also resulted in severe ultrastructural abnormalities, such as relaxed gap junctions between neighboring cardiomyocytes. Functionally, cardiovascular deletion of PSEN1 caused spontaneous mortality from birth to adulthood and led to diastolic heart dysfunction, including decreased volume of the left ventricle at the end‐systolic and end‐diastolic stages. Additionally, in a myocardial ischemia model, deletion of PSEN1 in the cardiovascular system first protected mice by inducing adaptive hypertrophy but ultimately resulted in severe heart failure. Furthermore, a collection of genes was abnormally expressed in the hearts of cardiac‐specific PSEN1 knockout mice. They were enriched in cell proliferation, calcium regulation, and so on. Taken together, dynamic regulation and abnormal function of PSEN1 underlie the pathogenesis of cardiovascular diseases due to ultrastructural abnormality of cardiomyocytes. Four types of genetically modified mice were established to determine the function of PSEN1 in cardiac development and pathology. Cardiovascular deletion of PSEN1 caused spontaneous mortality from birth to adulthood and led to diastolic heart dysfunction, including decreased volume of the left ventricle at the end‐systolic and end‐diastolic stages. Therefore, dynamic regulation and abnormal function of PSEN1 underlie the pathogenesis of cardiovascular diseases due to ultrastructural abnormality of cardiomyocytes.

  September 20, 2017   doi: 10.1002/jcp.26057   open full text
• Glucose stimulates intestinal epithelial crypt proliferation by modulating cellular energy metabolism.
  Weinan Zhou, Deepti Ramachandran, Abdelhak Mansouri, Megan J. Dailey.
  Journal of Cellular Physiology. September 19, 2017
  The intestinal epithelium plays an essential role in nutrient absorption, hormone release and barrier function. Maintenance of the epithelium is driven by continuous cell renewal by stem cells located in the intestinal crypts. The amount and type of diet influence this process and result in changes in the size and cellular make‐up of the tissue. The mechanism underlying the nutrient‐driven changes in proliferation is not known, but may involve a shift in intracellular metabolism that allows for more nutrients to be used to manufacture new cells. We hypothesized that nutrient availability drives changes in cellular energy metabolism of small intestinal epithelial crypts that could contribute to increases in crypt proliferation. We utilized primary small intestinal epithelial crypts from C57BL/6J mice to study 1) the effect of glucose on crypt proliferation, and 2) the effect of glucose on crypt metabolism using an extracellular flux analyzer for real‐time metabolic measurements. We found that glucose increased both crypt proliferation and glycolysis, and the glycolytic pathway inhibitor 2‐Deoxy‐D‐glucose (2‐DG) attenuated glucose‐induced crypt proliferation. Glucose did not enhance glucose oxidation, but did increase the maximum mitochondrial respiratory capacity, which may contribute to glucose‐induced increases in proliferation. Glucose activated Akt/HIF‐1α signaling pathway, which might be at least in part responsible for glucose‐induced glycolysis and cell proliferation. These results suggest that high glucose availability induces an increase in crypt proliferation by inducing an increase in glycolysis with no change in glucose oxidation. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26199   open full text
• Bone morphogenetic protein‐7 inhibits endothelial‐mesenchymal transition in pulmonary artery endothelial cell under hypoxia.
  Hongyue Zhang, Ying Liu, Lixin Yan, Wei Du, Xiaodan Zhang, Min Zhang, He Chen, Yafeng Zhang, Jianqiu Zhou, Hanliang Sun, Daling Zhu.
  Journal of Cellular Physiology. September 19, 2017
  Pulmonary artery hypertension (PAH) is characterized by structural changes in pulmonary arteries. Increased numbers of cells expressing α‐smooth muscle actin (α‐SMA) is a nearly universal finding in the remodeled artery. It has been confirmed endothelial‐to‐mesenchymal transition (EndoMT) may be a source of those α‐SMA–expressing cells. In addition, the EndoMT is reversible. Here, we show that under hypoxia, the expression of bone morphogenetic protein 7 (BMP‐7) was decreased both in vivo and in vitro. We also found that under normoxia, BMP‐7 deficiency induced spontaneous EndoMT and cell migration. The hypoxia‐induced EndoMT and cell migration were markedly attenuated after pretreatment with rh‐BMP‐7. Moreover, m‐TOR phosphorylation was involved in EndoMT and BMP‐7 suppressed hypoxia‐induced m‐TORC1 phosphorylation in pulmonary artery endothelial cells. Our results demonstrate that BMP‐7 attenuates the hypoxia‐induced EndoMT and cell migration by suppressing the m‐TORC1 signaling pathway. Our study revealed a novel mechanism underlying the hypoxia‐induced EndoMT in pulmonary artery endothelial cells and suggested a new therapeutic strategy targeting EndoMT for the treatment of pulmonary arterial hypertension. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26195   open full text
• Role of Viruses in Gastrointestinal cancer.
  Habibollah Mirzaei, Hossein Goudarzi, Gita Eslami, Ebrahim Faghihloo.
  Journal of Cellular Physiology. September 19, 2017
  Gastrointestinal cancers are a global public health problem, which represent a vast majority of all cancer‐caused deaths in both men and women. On the other hand, viral pathogens have been long implicated as etiological factors in the onset of certain human cancers, including gastrointestinal tumors. In this regard, Human Papilloma Virus (HPV), Epstein ‐ Barr Virus (EBV) and John Cunningham Virus (JCV) have been more strongly suggested to be involved in gastrointestinal carcinogenesis; so that, the association of HPV with oropharyngeal and anal cancers and also the association of EBV with gastric cancer have been etiologically confirmed by epidemiological and experimental investigations. Although, the association of other viruses is less evident, but may rely on co‐factors for their oncogenic roles. Therefore, to improve the prevention and treatment of these classes of cancer, their association with viral agents as potential risk factors should be investigated with care. In this respect, the present review has focused on the existing literature on the subject of viral involvement in gastrointestinal tumorgenesis, by covering and discussing various gastrointestinal cancers, corresponding viral agents and their oncogenic aspects and then summarizing evidences either supporting or rejecting a causal role of these pathogens in gastrointestinal malignancies. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26194   open full text
• Fenofibrate Reverses Changes Induced By High‐Fat Diet On Metabolism In Mice Muscle And Visceral Adipocytes.
  Flávia Frias, Karina Cunha e Rocha, Mariana de Mendonça, Gilson Massahiro Murata, Hygor Nunes Araujo4, Luís Gustavo Oliveira de Sousa, Érica de Souza, Sandro Massao Hirabara, Nayara Leite, Everardo Magalhães Carneiro, Rui Curi, Leonardo Reis Silveira, Alice Cristina Rodrigues.
  Journal of Cellular Physiology. September 19, 2017
  The effect of fenofibrate on the metabolism of skeletal muscle and visceral white adipose tissue of diet‐induced obese (DIO) mice was investigated. C57BL/6J male mice were fed either a control or high‐fat diet for eight weeks. Fenofibrate (50 mg/Kg b.w., daily) was administered by oral gavage during the last two weeks of the experimental period. Insulin‐stimulated glucose metabolism in soleus muscles, glucose tolerance test, insulin tolerance test, indirect calorimetry, lipolysis of visceral white adipose tissue, expression of miR‐103‐3p in adipose tissue and miR‐1a, miR‐133a/b, miR‐206, let7b‐5p, miR‐23b‐3p, miR‐29‐3p, miR‐143‐3p in soleus muscle, genes related to glucose and fatty acid metabolism in adipose tissue and soleus muscle, and proteins (phospho‐AMPKα2, Pgc1α, Cpt1b), intramuscular lipid staining, and activities of fatty acid oxidation enzymes in skeletal muscle were investigated. In DIO mice, fenofibrate prevented weight gain induced by HFD feeding by increasing energy expenditure; improved whole body glucose homeostasis, and in skeletal muscle, increased insulin dependent glucose uptake, miR‐1a levels, reduced intramuscular lipid accumulation, and phospho‐AMPKα2 levels. In visceral adipose tissue of obese mice, fenofibrate decreased basal lipolysis rate and visceral adipocytes hypertrophy, and induced the expression of Glut‐4, Irs1 and Cav‐1 mRNA and miR‐103‐3p suggesting a higher insulin sensitivity of the adipocytes. The evidence is presented herein that beneficial effects of fenofibrate on body weight, glucose homeostasis and muscle metabolism might be related to its action in adipose tissue. Moreover, fenofibrate regulates miR‐1a‐3p in soleus and miR‐103‐3p in adipose tissue, suggesting these microRNAs might contribute to fenofibrate beneficial effects on metabolism. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26203   open full text
• Distinct mechanisms of regulation of the ITGA6 and ITGB4 genes by RUNX1 in myeloid cells.
  Jessica L Phillips, Phillippa C Taberlay, Alexandra M Woodworth, Kristine Hardy, Kate H Brettingham‐Moore, Joanne L Dickinson, Adele F Holloway.
  Journal of Cellular Physiology. September 19, 2017
  Integrins are transmembrane adhesion receptors that play an important role in hematopoiesis by facilitating interactions between hematopoietic cells and extracellular matrix components of the bone marrow and hematopoietic tissues. These interactions are important in regulating the function, proliferation and differentiation of hematopoietic cells, as well as their homing and mobilization in the bone marrow. Not surprisingly altered expression and function of integrins plays a key role in the development and progression of cancer including leukemias. However, the regulation of integrin gene expression is not well characterized and the mechanisms by which integrin genes are disrupted in cancer remain unclear. Here we demonstrate for the first time that a key regulator of hematopoiesis, RUNX1, binds to and regulates the promoters of both the ITGA6 and ITGB4 genes in myeloid cells. The ITGA6 and ITGB4 integrin genes form the α6β4 integrin receptor. However our data indicates that RUNX1 functions differently at these two promoters. RUNX1 regulates ITGA6 through a consensus RUNX1 binding motif in its promoter. In contrast, although the ITGB4 promoter is also activated by RUNX1, it does so in the absence of a recognized consensus RUNX1 binding motif. Further, our data suggest that regulation of ITGB4 may involve interactions between the promoter and upstream regulatory elements. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26197   open full text
• FAM20C regulates osteoblast behaviors and intracellular signaling pathways in a cell‐autonomous manner.
  Chao Liu, Hua Zhang, Priyam Jani, Xiaofang Wang, Yongbo Lu, Nan Li, Jing Xiao, Chunlin Qin.
  Journal of Cellular Physiology. September 19, 2017
  Recent studies indicate that Family with sequence similarity 20 member C (FAM20C) catalyzes the phosphorylation of secreted proteins, and participates in a variety of biological processes, including cell proliferation, migration, mineralization and phosphate homeostasis. To explore the local influences of FAM20C on osteoblast, Fam20c‐deficient osteoblasts were generated by treating the immortalized Fam20cf/f osteoblasts with CMV‐Cre‐IRES‐EGFP lentivirus. Compared with the normal Fam20cf/f osteoblasts, the expression of Bone sialoprotein (Bsp), Osteocalcin (Ocn), Fibroblast growth factor 23 (Fgf23) and transcription factors that promote osteoblast maturation were up‐regulated in the Fam20c‐deficient osteoblasts. In contrast, the expression of Dental matrix protein 1 (Dmp1), Dentin sialophosphoprotein (Dspp), Osteopontin (Opn), type I Collagen a 1 (Col1a1) and Alkine phosphatase (Alp) were down‐regulated in the Fam20c‐deficient cells. These alterations disclosed the primary regulation of Fam20c on gene expression. The Fam20c‐deficient osteoblasts showed a remarkable reduction in the ability of forming mineralized nodules. However, supplements of extracellular matrix proteins extracted from the normal bone failed to rescue the reduced mineralization, suggesting that FAM20C may affect the biomineralization by the means more than local phosphorylation of extracellular matrix proteins and systemic phosphorus homeostasis. Moreover, although Fam20c deficiency had little impact on cell proliferation, it significantly reduced cell migration and lowered the levels of p‐Smad1/5/8, p‐Erk and p‐p38, suggesting that the kinase activity of FAM20C might be essential to cell mobility and the activity of BMP ligands. In summary, these findings provide evidences that FAM20C may regulate osteoblast maturation, migration, mineralization and BMP signaling pathways in a cell‐autonomous manner. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26200   open full text
• Loss Of Runx1 Is Associated With Aggressive Lung Adenocarcinomas.
  Jon Ramsey, Kelly Butnor, Zhihua Peng, Tim Leclair, Jos van der Velden, Gary Stein, Jane Lian, C. Matthew Kinsey.
  Journal of Cellular Physiology. September 19, 2017
  The mammalian runt‐related factor 1 (RUNX1) is a master transcription factor that regulates lineage specification of hematopoietic stem cells. RUNX1 translocations result in the development of myeloid leukemias. Recently, RUNX1 has been implicated as a tumor suppressor in other cancers. We postulated RUNX1 expression may be associated with lung adenocarcinoma etiology and/or progression. We evaluated the association of RUNX1 mRNA expression with overall survival data from The Cancer Genome Atlas (TCGA), a publically available database. Compared to high expression levels, Low RUNX1 levels from lung adenocarcinomas were associated with a worse overall survival (Hazard Ratio = 2.014 (1.042 to 3.730 95% confidence interval), log‐rank P = 0.035) compared to those that expressed high RUNX1 levels. Further immunohistochemical examination of 85 surgical specimens resected at the University of Vermont Medical Center identified that low RUNX1 protein expression was associated with larger tumors (P = 0.038). Gene expression network analysis was performed on the same subset of TCGA cases that demonstrated differential survival by RUNX1 expression. This analysis, which reveals regulatory relationships, showed that reduced RUNX1 levels were closely linked to upregulation of the transcription factor E2F1. To interrogate this relationship, RUNX1 was depleted in a lung cancer cell line that expresses high levels of RUNX1. Loss of RUNX1 resulted in enhanced proliferation, migration, and invasion. RUNX1 depletion also resulted in increased mRNA expression of E2F1 and multiple E2F1 target genes. Our data implicate loss of RUNX1 as driver of lung adenocarcinoma aggression, potentially through deregulation of the E2F1 pathway. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26201   open full text
• Oroxyloside inhibits angiogenesis through suppressing internalization of VEGFR2/Flk‐1 in endothelial cells.
  Kai Zhao, Xiaorui Li, Binyan Lin, Dawei Yang, Yuxin Zhou, Zhiyu Li, Qinglong Guo, Na Lu.
  Journal of Cellular Physiology. September 19, 2017
  Increasing flavonoids have been reported to possess anti‐angiogenic effects. Inhibition of angiogenesis plays a critical role in the treatment of cancer, especially in advanced metastatic cancer. In this study, we assessed the effect of Oroxylin A‐7‐glucuronide (Oroxyloside), a main metabolite of Oroxylin A, on angiogenesis in human endothelial cell‐like EA.hy926 cells. Oroxyloside suppressed the migration and tube formation of EA.hy926 cells. Meanwhile, microvessels sprouting from aortic rings and new blood vessels on the chicken chorioallantoic membrane (CAM) were also inhibited. Mechanism studies showed that Oroxyloside reduced the autophosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2/Flk‐1) while it up‐regulated the expression of R‐Ras and VE‐cadherin. In consequence, Oroxyloside inhibited the downstream Akt/MAPK/NF‐κB pathways and then decreased the nuclear translocation and DNA binding ability of NF‐κB. Furthermore, in‐vivo study showed that Oroxyloside exhibited a potential anti‐angiogenic effect in Matrigel plug assay and inhibited growth of xenografted tumors with low systemic toxicity, which could be ascribed to the inhibition of VEGFR2 internalization. Taken together, these results suggested that Oroxyloside could inhibit angiogenesis in vitro and in vivo via suppressing the internalization of VEGFR2 and might serve as a potential antitumor agent. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26198   open full text
• Regulating Osteogenesis and Adipogenesis in Adipose‐Derived Stem Cells by Controlling Underlying Substrate Stiffness.
  Tao Zhang, Shiyu Lin, Xiaoru Shao, Sirong Shi, Qi Zhang, Changyue Xue, Yunfeng Lin, Bofeng Zhu, Xiaoxiao Cai.
  Journal of Cellular Physiology. September 19, 2017
  Cells reside in a complex microenvironment (niche) in which the biochemical and biophysical properties of the extracellular matrix profoundly affect cell behavior. Extracellular stiffness, one important bio‐mechanical characteristic of the cell niche, is important in regulating cell proliferation, migration, and lineage specification. However, the mechanism by which mechanical signals guide osteogenic and adipogenic commitment of stem cells remains difficult to dissect. To explore this question, we generated a range of polydimethylsiloxane‐based matrices with differing degrees of stiffness that mimicked the stiffness seen in natural tissues and examined adipose stem cell morphology, spreading, vinculin expression and differentiation along the osteogenic and adipogenic pathways. Rigid matrices allowed broader cell spreading, faster growth rate and stronger expression of vinculin in adipose‐derived stem cells. In the presence of inductive culture media, stiffness‐dependent osteogenesis and adipogenesis of the adipose stem cells indicated that there was a combinatorial effect of biophysical and biochemical cues; no such lineage specification was observed in normal media. Osteogenic differentiation behavior showed a correlation with matrix rigidity, as well as with elevated expression of RhoA, ROCK‐1/‐2 and related proteins in the Wnt/β‐catenin pathway. The result provides a comprehensive understanding of how stem cells respond to the surrounding microenvironment and points to the fact that matrix stiffness is a critical element in biomaterial design and this will be an important advance in stem cell‐based tissue engineering. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26193   open full text
• Anti‐osteoporosis activity of Sanguinarine in preosteoblast MC3T3‐E1 cells and an ovariectomized rat model.
  Fuzhan Zhang, Jile Xie, Genlin Wang, Ge Zhang, Huilin Yang.
  Journal of Cellular Physiology. September 19, 2017
  Sanguinarine, a benzophenanthridine alkaloid, has been previously demonstrated to exert antimicrobial, anti‐inflammatory and anti‐tumor activities. A previous study has identified Sanguinarine as a potential drug candidate for osteoporosis treatment by computational bioinformatics analysis. This study further evaluated the effects of Sanguinarine on the differentiation of murine preosteoblast MC3T3‐E1 cells and its anti‐osteoporosis activity in an ovarietomized rat model. Sanguinarine treatment (0.25, 0.5, 1 and 2 µM) of MC3T3‐E1 cells significantly increased alkaline phosphatase (ALP) activity and the phoshporalyation of AMP‐activated protein kinase α subunit (AMPKα), but did not affect cell proliferation. The induction effects of Sanguinarine treatment (2 µM) on ALP activity, AMPKα phosphorylation, Smad1 phosphorylation and the expression of three osteoblast differentiation‐regulators (bone morphogenetic protein 2 [BMP2], osterix [OSX] and osteoprotegerin [OPG]) were partially reversed by Compound C treatment. More importantly, Sanguinarine treatment promoted bone tissue growth in an ovariectomized (OVX) osteoporosis rat model as evaluated by histological examination, micro‐CT analysis and serum parameter detection. In conclusion, these results indicate that Sanguinarine induces the differentiation of MC3T3‐E1 cells through the activation of the AMPK/Smad1 signaling pathway. Sanguinarine can stimulate bone growth in vivo and may be an effective drug for osteoporosis treatment. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26187   open full text
• Curcumin regulates proliferation, autophagy and apoptosis in gastric cancer cells by affecting PI3K and P53 signaling.
  Hongbing Fu, Changming Wang, Dejun Yang, Xin Zhang, Ziran Wei, Zhenxin Zhu, Jiapeng Xu, Zunqi Hu, Yu Zhang, Weimin Wang, Ronglin Yan, Qingping Cai.
  Journal of Cellular Physiology. September 19, 2017
  In this study, we aimed to investigate the effects of curcumin on cell activities of gastric cancer (GC), and the connection between curcumin and P53 as well as PI3K signaling. This study was conducted with two cell lines SGC‐7901 and BGC‐823, both were exposed to curcumin at the concentrations of 0 µM, 10 µM, 20 µM and 40 µM. MTT assay, flow cytometry (FCM) assay, transmission electron microscopy (TEM) were used to study the underlying mechanisms of curcumin in respective of proliferation, apoptosis and autophagy. Western blot assay was also employed to detect impacts of curcumin on tophosphatidylinositol‐3 kinase (PI3K) and P53 signaling pathways‐related proteins. MTT assay displayed that curcumin inhibited GC cell proliferation. FCM results indicated that curcumin induced the apoptosis of GC cells. TEM revealed that curcumin induced autophagy in GC cells. Western blot results showed that curcumin activated P53 signaling pathway and inhibited PI3K signaling pathway. Curcumin may inhibit proliferation and induce the autophagy and apoptosis in GC cells. Additionally, curcumin activated the P53 signaling pathway by up‐regulating P53 and P21, which also inhibited PI3K pathway through down‐regulating PI3K, p‐Akt and p‐mTOR. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26190   open full text
• Efficacy of different chemotherapy regimens in treatment of advanced or metastatic pancreatic cancer: a network meta‐analysis.
  Shu‐Hua Zhang, Gui‐Feng Liu, Xue‐Feng Li, Lin Liu, Shao‐Nan Yu.
  Journal of Cellular Physiology. September 19, 2017
  We performed a network meta‐analysis (NMA) to compare the short‐ and long‐term efficacy of Gemcitabine, Gemcitabine + S‐1 (tegafur), Gemcitabine + nab‐paclitaxel, Gemcitabine + Capecitabine, Gemcitabine + Cisplatin, FOLFIRINOX (oxaliplatin + irinotecan + fluorouracil + leucovorin), Gemcitabine + oxaliplatin, Gemcitabine + irinotecan, Gemcitabine + Exatecan, Gemcitabine + pemetrexed, Gemcitabine + 5‐FU and S‐1 in treating advanced or metastatic pancreatic cancer (PC). The odds radios (OR) or weighted mean difference (WMD) and surface under the cumulative ranking curves (SUCRA) were evaluated by a combination of direct evidence and indirect evidence. In total twenty studies were included in this paper. For short‐term efficacy, the overall response rate (ORR) was lower for patients treated with Gemcitabine compared with Gemcitabine + S‐1, Gemcitabine + Cisplatin, Gemcitabine + irinotecan and S‐1. The ORR for FOLFIRINOX was higher compared with Gemcitabine, Gemcitabine + Capecitabine and Gemcitabine + Cisplatin. The disease control rate (DCR) for Gemcitabine was lower compared with Gemcitabine + S‐1, Gemcitabine + Cisplatin and FOLFIRINOX. For long‐term efficacy, the 12‐month overall survival (OS) rate for FOLFIRINOX was higher compared with Gemcitabine, Gemcitabine + Capecitabine, Gemcitabine + Cisplatin, Gemcitabine + irinotecan, Gemcitabine + Exatecan and Gemcitabine + pemetrexed. The SUCRA revealed that FOLFIRINOX was relatively better in both short‐term and long‐term efficacy, while Gemcitabine was relatively poorer. In both short‐term and long‐term efficacy, FOLFIRINOX had the best short term and long term efficacy among the 12 chemotherapy regimens while efficacy of Gemcitabine was relatively poorer in the treatment of advanced or metastatic PC. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26183   open full text
• Long non‐coding RNA MALAT1 promotes oral squamous cell carcinoma development via microRNA‐125b/STAT3 axis.
  Shi‐Min Chang, Wei‐Wei Hu.
  Journal of Cellular Physiology. September 19, 2017
  Oral squamous cell carcinoma (OSCC), as the most common type of oral cancer, is responsible for almost 3% of all malignant tumors worldwide. Non‐coding RNAs such as lncRNAs and microRNAs have been involved in many cancers including OSCC. Recently, lncRNA metastasis‐associated lung adenocarcinoma transcript‐1 (MALAT1) has been reported to play an oncogenic role in OSCC metastasis. However, the underlying mechanism of MALAT1 in regulating OSCC progression remains unclear. The aim of this study was to investigate the specific role of MALAT1 in OSCC development. It was observed that MALAT1 was upregulated in OSCC cell lines. Inhibition of MALAT1 can prevent OSCC proliferation while overexpressing MALAT1 promoted OSCC progression. In addition, bioinformatics search was used to identify that miR‐125b was a direct target of MALAT1, which indicated a negative correlation between MALAT1 and miR‐125b. Besides these, STAT3 was predicted as a binding target of miR‐125b in OSCC. Overexpression of MALAT1 was able to suppress the tumor inhibitory effect of miR‐125b mimics via upregulating STAT3. Moreover, the function of MALAT1 in OSCC development was further investigated by using in vivo assays. The established nude mice models revealed that downregulated MALAT1 greatly inhibited OSCC tumor growth and reversely upregualated MALAT1 promoted OSCC development via miR‐125b/STAT3 axis respectively. In conclusion, MALAT1 can function as a competing endogenous RNA (ceRNA) to modulate STAT3 expression by absorbing miR‐125b in OSCC and could be used as a novel therapeutic target in OSCC diagnosis and treatment. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26185   open full text
• Long non‐coding RNA LINC00968 acts as oncogene in NSCLC by activating the Wnt signaling pathway.
  Yíng Wang, Jun Zhou, Yu‐Jun Xu, Hai‐Bo Hu.
  Journal of Cellular Physiology. September 19, 2017
  Long non‐coding RNAs (lncRNAs) have played critical roles in a variety of cancers, including non‐small cell lung cancer (NSCLC). In our study, we focused on the biological function and clinical significance of lncRNA LINC00968 in NSCLC. It was indicated that LINC00968 was significantly increased in LUAD tissues, LUSC tissues and NSCLC cells compared to their corresponding controls. Inhibition of LINC00968 was able to repress NSCLC growth, migration and invasion in vitro while upregulation of LINC00968 reversed this process. Additionally, downregulation of LINC00968 induced apoptosis capacity of A549 cell. Apoptosis‐related proteins BCL‐2 were decreased and BAX was increased by knockdown of LINC00968 respectively. Meanwhile we observed that Wnt signaling pathway was involved in the LINC00968‐induced NSCLC progression. Finally, in vivo tumor xenografts were established using A549 cells to detect the function of LINC00968 in NSCLC tumorigenesis. Silencing LINC00968 greatly inhibited NSCLC tumor progression, which was consistent with the in vitro tests. In conclusion, we have uncovered that LINC00968 could be regarded as a novel prognostic biomarker and therapeutic target in NSCLC diagnosis and treatment. This article is protected by copyright. All rights reserved

  September 19, 2017   doi: 10.1002/jcp.26186   open full text
• Primary familial brain calcification with a novel SLC20A2 mutation: Analysis of PiT‐2 expression and localization.
  Ilaria Taglia, Patrizia Formichi, Carla Battisti, Giulia Peppoloni, Melissa Barghigiani, Alessandra Tessa, Antonio Federico.
  Journal of Cellular Physiology. September 18, 2017
  Primary familial brain calcification (PFBC) is an autosomal dominant rare disorder characterized by bilateral and symmetric brain calcifications, and neuropsychiatric manifestations. Four genes have been linked to PFBC: SLC20A2, PDGFRB, PDGFB, and XPR1. In this study, we report molecular and clinical data of a PFBC patient carrying a novel SLC20A2 mutation and we investigate the impact of the mutation on PiT‐2 expression and function. Sanger sequencing of SLC20A2, PDGFRB, PDGFB, XPR1 led to the identification of a novel duplication of twelve nucleotides (c.1876_1887dup/ p.Trp626_Thr629dup) in SLC20A2 gene. SLC20A2 encodes for a cell membrane transporter (PiT‐2) involved in maintenance of inorganic phosphate homeostasis. We performed an analysis of expression and functionality of PiT‐2 protein in patient primary cultured fibroblasts. In patient fibroblasts, the mutation does not affect PiT‐2 expression but alter sub‐cellular localization. The Pi‐uptake assay revealed a less Pi depletion in patient than in control fibroblasts, suggesting that SLC20A2 duplication may impair Pi internalization. This is the first study reporting sub‐cellular expression analysis of mutant PiT‐2 in primary cultured fibroblasts from a PFBC patient, showing that p.Trp626_Thr629dup in SLC20A2 alters PiT‐2 sub‐cellular localization and reduces Pi‐uptake, leading to onset of PFBC in our patient. Primary familial brain calcification (PFBC) is an autosomal dominant rare disorder characterized by brain calcifications and neuropsychiatric manifestations. In this study, we report molecular and clinical data of a PFBC patient carrying a novel duplication of twelve nucleotides (c.1876_1887dup/ p.Trp626_Thr629dup) in SLC20A2 gene. SLC20A2 encodes for a cell membrane transporter (PiT‐2) involved in maintenance of inorganic phosphate homeostasis. The mutation does not affect PiT‐2 expression but alter sub‐cellular localization and impair Pi internalization.

  September 18, 2017   doi: 10.1002/jcp.26104   open full text
• Triptolide prevents extracellular matrix accumulation in experimental diabetic kidney disease by targeting microRNA‐137/Notch1 pathway.
  Fei Han, Shanshan Wang, Yunpeng Chang, Chunjun Li, Juhong Yang, Zhe Han, Baocheng Chang, Bei Sun, Liming Chen.
  Journal of Cellular Physiology. September 13, 2017
  MicroRNAs (miRNAs) are involved in multiple biological functions via suppressing target genes. Triptolide is a monomeric compound isolated from a traditional Chinese herb, which exerts protective roles in many kinds of glomerular diseases. However, our understanding of the triptolide effect on miRNAome is still limited. In this study, we found that triptolide significantly decreased albuminuria and improved glomerulosclerosis in rats with diabetic kidney disease (DKD). And triptolide also inhibited extracellular matrix (ECM) protein accumulation and the notch1 pathway activation under diabetic conditions. MiR‐137 was significantly decreased in the HG (high glucose)‐treated HRMCs and in the kidney tissues of the diabetic rats, but was upregulated by triptolide. In addition, overexpression of miR‐137 exerted similar effects to those of triptolide, while miR‐137 inhibition aggravated ECM protein accumulation. Luciferase reporter assay results demonstrated that miR‐137 directly targets Notch1. Furthermore, the miR‐137‐dependent effects were due to Notch1 suppression that in turn inhibited ECM protein expression, key mediators of glomerulosclerosis. Finally, downregulation of miR‐137 reversed the ECM inhibition role of triptolide in HG cultured HRMCs. Taken together, these findings indicate that triptolide is a potential therapeutic option for DKD and that miR‐137/Notch1 pathway play roles in the anti‐glomerulosclerosis mechanism of triptolide. Diabetic condition decreases the miR‐137 levels in renal mesangial cells. MiR‐137 decreases the expression of Col IV and FN by targeting Notch1. On the other hand, triptolide may protect DKD against glomerulosclerosis by targeting miR‐137/Notch1 pathway.

  September 13, 2017   doi: 10.1002/jcp.26092   open full text
• Double sex and mab‐3 related transcription factor 1 regulates differentiation and proliferation in dairy goat male germline stem cells.
  Yudong Wei, Shufang Cai, Fanglin Ma, Ying Zhang, Zhe Zhou, Shuanshuan Xu, Mengfei Zhang, Sha Peng, Jinlian Hua.
  Journal of Cellular Physiology. September 13, 2017
  The protein encoded by double sex and mab‐3 related transcription factor 1 (Dmrt1) gene contains a double sex/mab‐3 domain, which was considered as one of the most conservative structures in sex determination. However, its effect on spermatogenesis of dairy goat spermatogonial stem cells (SSCs) remains to be clarified. For the first time, the roles of Dmrt1 in spermatogenesis of livestock are highlighted. Here, we investigated the expression pattern of Dmrt1 in the testes of dairy goats. Dmrt1 primarily located in undifferentiated SSCs. Moreover, Dmrt1 enhanced differentiation and proliferation of mGSCs. On the contrary, the level of meiosis was down‐regulated, as Dmrt1 determines whether SSCs undergo mitosis and spermatogonial differentiation or meiosis. In the busulfan‐treated mice testes, Dmrt1 repair germ cell damage was emphasized as well. Our results exposed that Dmrt1 maintenance mGSCs in two ways: facilitating proliferation and self‐renewal of SSCs; and reducing the inflammatory response caused by reproductive injury. These findings identify a central role for Dmrt1 in controlling population stability and injury restoring of SSCs. For the first time, the roles of Dmrt1 in spermatogenesis of livestock are highlighted. Dmrt1 primarily located in undifferentiated SSCs, moreover, Dmrt1 enhanced differentiation and proliferation of mGSCs. Our results exposed that Dmrt1 maintenance mGSCs in two ways: facilitating proliferation and self‐renewal of SSCs; and reducing the inflammatory response caused by reproductive injury.

  September 13, 2017   doi: 10.1002/jcp.26129   open full text
• Iodixanol versus iopromide in cancer patients: Evidence from a randomized clinical trial.
  Irene Terrenato, Francesca Sperati, Felice Musicco, Anna F. Pozzi, Annunziata di Turi, Mauro Caterino, Elisabetta de Lutio di Castelguidone, Sergio V. Setola, Massimo Bellomi, Carlo E. Neumaier, Laura Conti, Giovanni Cigliana, Roberta Merola, Anna Antenucci, Giulia Orlandi, Antonio Giordano, Maddalena Barba, Stefano Canitano.
  Journal of Cellular Physiology. September 12, 2017
  To assess the safety profile of iso‐osmolar contrast medium (CM) versus low osmolar CM in cancer patients with an estimated glomerular filtration rate (eGFR) >60 ml/min. In this multicenter, blind trial of patients seeking a chest‐abdomen‐pelvis contrast enhanced computed tomography (CT) with iodated CM, participants were centrally randomized to iodixanol or iopromide. Contrast induced nephropathy (CIN) at 24 and/or 72 hr were our primary outcomes. We further considered irreversible CIN, average eGFR percentage variation (%Δ), and adverse events (AEs). Overall, 607 patients were enrolled. Among them, 497 eligible patients were randomized to iodixanol (N: 247) or iopromide (N: 250). No differences emerged by descriptive characteristics. Seven and 3 CIN at 24 hr (p = 0.34) and 8 and 2 CIN at 72 hr (p = 0.11) occurred in the iopromide and iodixanol group, respectively. Within the subgroup of individual patients who developed CIN (N: 17), the event rate was higher in the iopromide arm (p = 0.045). No cases of permanent CIN or significant differences in terms of AEs or GFR %Δ were observed. Our results suggest a more favorable safety profile of iodixanol versus iopromide. Adequately sized trials with similar design are warranted to confirm our findings and clarify the underlying biological mechanisms. 1) This is a multicenter, blind trial of patients seeking a chest‐abdomen‐pelvis CT with iodated CM. 2) Aim of the study was to assess the safety profile of iso‐osmolar contrast medium (CM) versus low osmolar CM in cancer patients with an estimated glomerular filtration rate (eGFR) >60 ml/min. 3) No cases of permanent CIN or significant differences in terms of adverse events were observed.

  September 12, 2017   doi: 10.1002/jcp.26132   open full text
• The ubiquitin ligase TRIM56 inhibits ovarian cancer progression by targeting vimentin.
  Lei Zhao, Ping Zhang, Xiao‐jie Su, Bing Zhang.
  Journal of Cellular Physiology. September 12, 2017
  Tumor metastasis is responsible for 90% of all cancer‐related deaths. Epithelial to mesenchymal transition (EMT) is an important prerequisite for tumor metastasis. One of the important mediators of EMT and cancer progression in ovarian cancer is the vimentin protein. The objective of the current study was to evaluate the molecular mechanism that regulates vimentin expression in ovarian cancer cells. Vimentin was robustly induced in the ovarian cancer cell line SKOV‐3 compared to normal ovarian epithelial cell line Moody and the induction was not due to transcriptional upregulation. Treatment with the proteasomal inhibitor MG‐132 revealed that vimentin is actively degraded by the proteasome in Moody cells and stabilized in the SKOV‐3 cell line. Mass spectrometric analysis of vimentin immunoprecipitate of MG‐132 treated Moody cells revealed candidate ubiquitin ligases associated with vimentin. RNAi mediated silencing of the candidate ubiquitin in Moody cells and concurrent overexpression of the candidate ubiquitin ligases in SKOV‐3 confirmed that TRIM56 is the ubiquitin ligase that is degrading vimentin in Moody cells. RNAi mediated silencing of TRIM56 in Moody cells and ectopic overexpression of TRIM56 in SKOV‐3 cells, respectively, significantly up‐ and down‐regulated in vitro migration and invasion in these cells. Analysis of TRIM56 transcript level and vimentin protein expression in 25 patients with ovarian carcinoma confirmed an inverse correlation between TRIM56 and vimentin expression. Cumulatively, our data reveals for the first time a novel post‐translational regulatory mechanism of regulating vimentin expression, EMT, and metastatic progression in ovarian cancer cells. RNAi mediated silencing of TRIM56 in Moody cells and ectopic overexpression of TRIM56 in SKOV‐3 cells, respectively, significantly up‐ and down‐regulated in vitro migration and invasion in these cells. Analysis of TRIM56 transcript level and vimentin protein expression in 25 patients with ovarian carcinoma confirmed an inverse correlation between TRIM56 and vimentin expression.

  September 12, 2017   doi: 10.1002/jcp.26114   open full text
• mSEL‐1L deficiency affects vasculogenesis and neural stem cell lineage commitment.
  Marina Cardano, Giuseppe R. Diaferia, Luciano Conti, Simona Baronchelli, Alessandro Sessa, Vania Broccoli, Andrea Barbieri, Pasquale De Blasio, Ida Biunno.
  Journal of Cellular Physiology. September 12, 2017
  mSEL‐1L is a highly conserved ER‐resident type I protein, involved in the degradation of misfolded peptides through the ubiquitin–proteasome system (UPS), a pathway known to control the plasticity of the vascular smooth muscle cells (VSMC) phenotype and survival. In this article, we demonstrate that mSEL‐1L deficiency interferes with the murine embryonic vascular network, showing particular irregularities in the intracranic and intersomitic neurovascular units and in the cerebral capillary microcirculation. During murine embryogenesis, mSEL‐1L is expressed in cerebral areas known to harbor progenitor neural cells, while in the adult brain the protein is specifically restricted to the stem cell niches, co‐localizing with Sox2 and Nestin. Null mice are characterized by important defects in the development of telenchephalic regions, revealing conspicuous aberration in neural stem cell lineage commitment. Moreover, mSEL‐1L depletion in vitro and in vivo appears to affect the harmonic differentiation of the NSCs, by negatively influencing the corticogenesis processes. Overall, the data presented suggests that the drastic phenotypic characteristics exhibited in mSEL‐1L null mice can, in part, be explained by the negative influence it plays on Notch1 signaling pathway. mSEL‐1L depletion in vitro and in vivo appears to affect the harmonic differentiation of the NSCs, by negatively influencing the corticogenesis processes. The data presented suggests that the drastic phenotypic characteristics exhibited in mSEL‐1L null mice can, in part, be explained by the negative influence it plays on Notch1 signaling pathway.

  September 12, 2017   doi: 10.1002/jcp.26153   open full text
• Bifunctional role of ephrin A1‐Eph system in stimulating cell proliferation and protecting cells from cell death through the attenuation of ER stress and inflammatory responses in bovine mammary epithelial cells.
  Minkyung Kang, Wooyoung Jeong, Hyocheol Bae, Whasun Lim, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. September 11, 2017
  Structural and functional development of the mammary gland is constant in the mammary gland life cycle. Eph receptors and their ligands, ephrins, control events through cell‐to‐cell interactions during embryonic development, and adult tissue homeostasis; however, little information on participation of ephrin A1, a representative ligand of the Eph receptor, in the development and function of normal mammary glands is known. In this study, we demonstrated functional effects of the ephrin A1‐Eph system and mechanisms of its action on bovine mammary epithelial (MAC‐T) cells. The in vitro cultured MAC‐T cells expressed the ephrin A1 ligand and EphA1, A2, A4, A7, and A8 among the eight members of the Eph A family. Our results revealed that ephrin A1 induced MAC‐T cell cycle progression and stimulated cell proliferation with abundant expression of nucleic PCNA and cyclin D1 proteins. Additionally, ephrin A1 induced activation of intracellular signaling molecules involved in PI3 K/AKT and MAPK signaling, and the proliferation‐stimulating effect of ephrin A1 was mediated by activation of these pathways. Furthermore, ephrin A1 influenced expression and activation of various ER stress‐related proteins and protected MAC‐T cells from stress‐induced cell death. Finally, ephrin A1 alleviated LPS‐induced cell death through down‐regulation of inflammatory cytokines. In conclusion, the results of this study suggest that the Eph A‐ephrin A1 system is a positive factor in the increase and maintenance of epithelial cells in mammary glands of cows; the signaling system contributes to development, remodeling, and functionality of normal mammary glands and could overcome mastitis in cows and other mammals. The Eph A‐ephrin A1 system is a positive factor in the increase and maintenance of epithelial cells in mammary glands of cows; the signaling system contributes to development, remodeling, and functionality of normal mammary glands and could overcome mastitis in cows and other mammals.

  September 11, 2017   doi: 10.1002/jcp.26131   open full text
• Gap Junctional Intercellular Communication in Adipose‐derived Stromal/Stem Cells is Cell Density‐dependent and Positively Impacts Adipogenic Differentiation.
  Wiesner Miriam, Berberich Oliver, Hoefner Christiane, Blunk Torsten, Bauer‐Kreisel Petra.
  Journal of Cellular Physiology. September 09, 2017
  Adipose‐derived stromal/stem cells (ASCs) represent a widely used cell source with multi‐lineage differentiation capacity in approaches for tissue engineering and regenerative medicine. Despite the multitude of literature on their differentiation capacity, little is reported about the physiological properties contributing to and controlling the process of lineage differentiation. Direct intercellular communication between adjacent cells via gap junctions has been shown to modulate differentiation processes in other cell types, with connexin 43 (Cx43) being the most abundant isoform of the gap junction‐forming connexins. Thus, in the present study we focused on the expression of Cx43 and gap junctional intercellular communication (GJIC) in human ASCs, and its significance for adipogenic differentiation of these cells. Cx43 expression in ASCs was demonstrated histologically and on the gene and protein expression level, and was shown to be greatly positively influenced by cell seeding density. Functionality of gap junctions was proven by dye transfer analysis in growth medium. Adipogenic differentiation of ASCs was shown to be also distinctly elevated at higher cell seeding densities. Inhibition of GJIC by 18α‐glycyrrhetinic acid (AGA) significantly compromised adipogenic differentiation, as demonstrated by histology, triglyceride quantification, and adipogenic marker gene expression. Flow cytometry analysis showed a lower proportion of cells undergoing adipogenesis when GJIC was inhibited, further indicating the importance of GJIC in the differentiation process. Altogether, this study demonstrates the impact of direct cell‐cell communication via gap junctions on the adipogenic differentiation process of ASCs, and may contribute to further integrate direct intercellular crosstalk in rationales for tissue engineering approaches. This article is protected by copyright. All rights reserved

  September 09, 2017   doi: 10.1002/jcp.26178   open full text
• Systematic‐analysis of mRNA Expression Profiles in Skeletal Muscle of Patients with Type II Diabetes: the Glucocorticoid was Central in Pathogenesis.
  Kan Shao, Li‐Sha Shen, Hui‐Hua Li, Shan Huang, Yong Zhang.
  Journal of Cellular Physiology. September 08, 2017
  Since the past thirty years, the prevalence of diabetes has more than doubled, making it an urgent challenge globally. We carried out systematic analysis with the public data of mRNA expression profiles in skeletal muscle to study the pathogenesis, since insulin resistance in the skeletal muscle is an early feature. We utilized three GEO datasets, containing total 60 cases and 63 normal samples. After the background removal, R package QC was utilized to finish the preprocessing of datasets. We obtained a dataset containing 2481 genes and 123 samples after the pre‐processing. Quantitative quality control measures were calculated to represent the quality of these datasets. MetaDE package provides functions for conducting different systematic analysis methods for differential expression analysis. The GO term enrichment was carried out using PANTHER. Protein‐protein interactions, drug‐gene interactions and genetic association of the identified differentially expressed genes were analyzed using STRING v10.0 online tool, DGIdb and the Genetic Association Database, respectively. The datasets had good performances on IQC and EQC, which suggested that the datasets had good internal and external quality. Totally 96 differentially expressed genes were detected using 0.01 as cutoff of AW. The enriched GO terms were mainly associated with the response to glucocorticoid. There were seven genes involving in the gluconeogenesis were differentially expressed, which might be the potential treatment target for this disease. The closely connected networks and potential targets of existed drugs suggested that some of the drugs might be applied to the treatment of diabetes as well. This article is protected by copyright. All rights reserved

  September 08, 2017   doi: 10.1002/jcp.26174   open full text
• Novel role for the testis‐enriched HSPA2 protein in regulating epidermal keratinocyte differentiation.
  Agnieszka Gogler‐Pigłowska, Katarzyna Klarzyńska, Damian R. Sojka, Anna Habryka, Magdalena Głowala‐Kosińska, Marcin Herok, Mariusz Kryj, Monika Halczok, Zdzisław Krawczyk, Dorota Scieglinska.
  Journal of Cellular Physiology. September 07, 2017
  HSPA2, a poorly characterized member of the HSPA (HSP70) chaperone family, is a testis‐enriched protein involved in male germ cell differentiation. Previously, we revealed that HSPA2 is present in human stratified epithelia, including epidermis, however the contribution of this protein to epithelial biology remained unknown. Here, we show for the first time that HSPA2 is expressed in basal epidermal keratinocytes, albeit not in keratinocytes exhibiting features attributed to primitive undifferentiated progenitors, and participates in the keratinocyte differentiation process. We found that HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. We also shown that lentiviral‐mediated shRNA silencing of HSPA2 expression in HaCaT cells caused a set of phenotypic changes characteristic for keratinocytes committed to terminal differentiation such as reduced clonogenic potential, impaired adhesiveness and increased basal and confluency‐induced expression of differentiation markers. Moreover, the fraction of undifferentiated cells that rapidly adhered to collagen IV was less numerous in HSPA2‐deficient cells than in the control. In a 3D reconstructed human epidermis model, HSPA2 deficiency resulted in accelerated development of a filaggrin‐positive layer. Collectively, our results clearly show a link between HSPA2 expression and maintenance of keratinocytes in an undifferentiated state in the basal layer of the epidermis. It seems that HSPA2 could retain keratinocytes from premature entry into the terminal differentiation process. Overall, HSPA2 appears to be necessary for controlling development of properly stratified epidermis and thus for maintenance of skin homeostasis. In human skin HSPA2 is preferentially expressed in basal epidermal keratinocytes. HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. HSPA2 participates in the keratinocyte differentiation process.

  September 07, 2017   doi: 10.1002/jcp.26142   open full text
• DHQZ‐17, a potent inhibitor of the transcription factor HNF4A, suppresses tumorigenicity of head and neck squamous cell carcinoma in vivo.
  Shilpa Tentu, Kumarswamyreddy Nandarapu, Prakash Muthuraj, Kesavan Venkitasamy, Ganesh Venkatraman, Suresh K. Rayala.
  Journal of Cellular Physiology. September 07, 2017
  A series of 2, 3‐dihydroquinazolinone derivatives were synthesized, characterized and their anticancer activity was determined. Among the compounds synthesized and screened, one compound (17) showed potent anticancer activity against human head and neck squamous cell carcinoma cell line, SCC131 and was non‐toxic to normal cells. The compound inhibited the growth of SCC131 cells, with an IC50 of 1.75 μM, triggered apoptotic mode of cell death and caused tumor regression of SCC131 tumor xenografts in athymic mice. To decipher the target for the lead compound, a high throughput qPCR array was performed. Results showed that the compound 17, inhibited the expression of a vital transcription factor HNF4A, involved in regulation of metabolic pathways. Thus, the present work has identified a lead compound 17, with potent anticancer activity, minimal normal cell toxicity and a plausible target and hence definitely holds future prospects as an anticancer agent. A novel small molecule inhibitor for oral cancer.

  September 07, 2017   doi: 10.1002/jcp.26139   open full text
• CRISPR editing in biological and biomedical investigation.
  Jiaojiao Huang, Yanfang Wang, Jianguo Zhao.
  Journal of Cellular Physiology. September 07, 2017
  Recently, clustered regularly interspaced short palindromic repeats (CRISPR) based genomic editing technologies have armed researchers with powerful new tools to biological and biomedical investigations. To further improve and expand its functionality, natural, and engineered CRISPR associated nine proteins (Cas9s) have been investigated, various CRISPR delivery strategies have been tested and optimized, and multiple schemes have been developed to ensure precise mammalian genome editing. Benefiting from those in‐depth understanding and further development of CRISPR, versatile CRISPR‐based platforms for genome editing have been rapidly developed to advance investigations in biology and biomedicine. In biological research area, CRISPR has been widely adopted in both fundamental and applied research fields, such as accurate base editing, transcriptional regulation, and genome‐wide screening. In biomedical research area, CRISPR has also shown its extensive applicability in the establishment of animal models for genetic disorders especially those large animals and non‐human primates models, and gene therapy to combat virus infectious diseases, to correct monogenic disorders in vivo or in pluripotent cells. In this prospect article, after highlighting recent developments of CRISPR systems, we outline different applications and current limitations of CRISPR use in biological and biomedical investigation. Finally, we provide a perspective for future development and potential risks of this multifunctional technology. In this prospect article, after highlighting recent developments of CRISPR systems, we outline different applications and current limitations of CRISPR use in biological and biomedical investigation. Finally, we provide a perspective for future development and potential risks of this multifunctional technology.

  September 07, 2017   doi: 10.1002/jcp.26141   open full text
• Potential role of the Jagged1/Notch1 signaling pathway in the endothelial‐myofibroblast transition during BLM‐induced pulmonary fibrosis.
  Qian Yin, Weihua Wang, Guangbin Cui, Linfeng Yan, Song Zhang.
  Journal of Cellular Physiology. September 07, 2017
  Endothelial cell myofibroblast transition (EndoMT) is found during the process of bleomycin (BLM)‐induced pulmonary fibrosis in rats, and plays a very important role in sustaining inflammation and collagen secretion. Moreover, some studies have suggested that the Notch1 signaling pathway may be involved in the expression of α‐smooth muscle actin (α‐SMA) in pulmonary microvascular endothelial cells (PMVECs), a protein marker of EndoMT. Therefore, we aimed to investigate the expression level of α‐SMA and Notch1‐related signaling molecules in PMVECs from BLM‐induced rats and determine the relationship between the Notch1 signaling pathway and the expression of α‐SMA in PMVECs. We found that the expression levels of α‐SMA, Notch1, and Jagged1 were upregulated, while the expression levels of Dll4 were downregulated. Furthermore, there was a positive correlation between the expression of Jagged1 and the α‐SMA proteins in PMVECs, and NF‐κB was downregulated by decreasing the expression of Jagged1. In conclusion, the Jagged1/Notch1 signaling pathway is activated in PMVECs during the pathogenesis of BLM‐induced pulmonary fibrosis in rats, and it may induce α‐SMA expression via a non‐canonical pathway involving NF‐κB as the target molecule. The precise mechanism and the molecules involved in this signaling pathway need to be further elucidated. The Jagged1/Notch1 signaling pathway is activated in PMVECs during the pathogenesis of BLM‐induced pulmonary fibrosis in rats, and it may induce α‐SMA expression via a non‐canonical pathway involving NF‐κB as the target molecule.

  September 07, 2017   doi: 10.1002/jcp.26122   open full text
• Cyanidin Chloride inhibits ovariectomy‐induced osteoporosis by suppressing RANKL‐mediated osteoclastogenesis and associated signaling pathways.
  Jianwen Cheng, Lin Zhou, Qian Liu, Jennifer Tickner, Zhen Tan, Xiaofeng Li, Mei Liu, Xixi Lin, Tao Wang, Nathan J. Pavlos, Jinmin Zhao, Jiake Xu.
  Journal of Cellular Physiology. September 07, 2017
  Over‐production and activation of osteoclasts is a common feature of osteolytic conditions such as osteoporosis, tumor‐associated osteolysis, and inflammatory bone erosion. Cyanidin Chloride, a subclass of anthocyanin, displays antioxidant and anti‐carcinogenesis properties, but its role in osteoclastic bone resorption and osteoporosis is not well understood. In this study, we showed that Cyanidin Chloride inhibits osteoclast formation, hydroxyapatite resorption, and receptor activator of NF‐κB ligand (RANKL)‐induced osteoclast marker gene expression; including ctr, ctsk, and trap. Further investigation revealed that Cyanidin Chloride inhibits RANKL‐induced NF‐κB activation, suppresses the degradation of IκB‐α and attenuates the phosphorylation of extracellular signal‐regulated kinases (ERK). In addition, Cyanidin Chloride abrogated RANKL‐induced calcium oscillations, the activation of nuclear factor of activated T cells calcineurin‐dependent 1 (NFATc1), and the expression of c‐Fos. Further, we showed that Cyanidin Chloride protects against ovariectomy‐induced bone loss in vivo. Together our findings suggest that Cyanidin Chloride is capable of inhibiting osteoclast formation, hydroxyapatite resorption and RANKL‐induced signal pathways in vitro and OVX‐induced bone loss in vivo, and thus might have therapeutic potential for osteolytic diseases. Cyanidin Chloride inhibits osteoclast formation, hydroxyapatite resorption, and receptor activator of NF‐kB ligand (RANKL)‐induced osteoclast marker gene expression; including ctr, ctsk, and trap. Cyanidin Chloride inhibits RANKL‐induced NF‐kB activation, suppresses the degradation of IkB‐a and attenuates the phosphorylation of extracellular signal‐regulated kinases (ERK). Cyanidin Chloride also abrogates RANKL‐induced calcium oscillations, the activation of nuclear factor of activated T cells calcineurin‐dependent 1 (NFATc1), and the expression of c‐Fos. Cyanidin Chloride protects against ovariectomy‐induced bone loss in vivo. Cyanidin Chloride is capable of inhibiting osteoclast formation, hydroxyapatite resorption and RANKL‐induced signal pathways in vitro and OVX‐induced bone loss in vivo, and thus might have therapeutic potential for osteolytic diseases.

  September 07, 2017   doi: 10.1002/jcp.26126   open full text
• Increased expression of PD‐L1 and PD‐L2 in dermal fibroblasts from alopecia areata mice.
  Yunyuan Li, Ruhi T. Kilani, Mohammadreza Pakyari, Gigi Leung, Layla Nabai, Aziz Ghahary.
  Journal of Cellular Physiology. September 04, 2017
  Alopecia areata (AA) is a common autoimmune disorder affecting millions of people worldwide, which manifests as a sudden, non‐scarring hair loss. The expression of a pro‐inflammatory cytokine, interferon‐gamma (INF‐γ), has been well established to be involved in the development of AA. As IFN‐γ and other cytokines are also known to up‐regulate programmed cell death ligand 1 and 2 (PD‐L1 and PD‐L2), which both negatively control immune responses, we asked whether or not a high number of infiltrated T cells, seen in AA lesions, can modulate the expression of PD‐L1 and PD‐L2 in skin cells. From a series of experiments, we showed that a significantly higher number of PD‐L1 or PD‐L2 positive cells affect the skin in AA mice, compared to the skin of non‐AA mice. The number of PD‐L1 positive cells was well correlated with the number of infiltrated T cells, especially CD8+ T cells. We also found that the expression of PD‐L1 and PD‐L2 was co‐localized with type 1 pro‐collagen, CD90 and vimentin, which are biomarkers for dermal fibroblasts. Further studies revealed that releasable factors from activated, but not inactivated, lymphocytes significantly increase the expressions of both PD‐L1 and PD‐L2 in cultured dermal fibroblasts. In conclusion, our findings suggest that the expression of PD‐L1 and PD‐L2 in dermal fibroblasts is up‐regulated by activated T cells in AA‐affected skin, and as such, these regulatory molecules may not exert a negative control of the immune activation seen in AA lesions. (i) The highly expression of PD‐L1 and PD‐L2 were found in AA‐affected skin in a mouse mode; (ii) we identified dermal fibroblasts as the skin cells to express PD‐L1 and PD‐L2 in AA‐affected skin and; (iii) the expressibon of PD‐L1 and PD‐L2 in dermal fibrolasts were regulated by the releasable factors from activated lymphocytes.

  September 04, 2017   doi: 10.1002/jcp.26134   open full text
• The ubiquitin ligase SCFFBXW7α promotes GATA3 degradation.
  Nan Song, Cheng Cao, Yiman Tang, Liyuan Bi, Yong Jiang, Yongsheng Zhou, Xin Song, Ling Liu, Wenshu Ge.
  Journal of Cellular Physiology. September 04, 2017
  GATA3 is a key transcription factor in cell fate determination and its dysregulation has been implicated in various types of malignancies. However, how the abundance and function of GATA3 are regulated remains unclear. Here, we report that GATA3 is physically associated with FBXW7α, and FBXW7α destabilizes GATA3 through assembly of a SKP1‐CUL1‐F‐box E3 ligase complex. Importantly, we showed that FBXW7α promotes GATA3 ubiquitination and degradation in a GSK3 dependent manner. Furthermore, we demonstrated that FBXW7α inhibits breast cancer cells survival through destabilizing GATA3, and the expression level of FBXW7α is negatively correlated with that of GATA3 in breast cancer samples. This study indicated that FBXW7α is a critical negative regulator of GATA3 and revealed a pathway for the maintenance of GATA3 abundance in breast cancer cells. In this study, we aimed to understand how the abundance and function of transcription factor GATA3 are regulated. We revealed that FBXW7α interacts with and promotes turnover of GATA3, and FBXW7α suppresses breast cancer cell survival by degrading GATA3.

  September 04, 2017   doi: 10.1002/jcp.26108   open full text
• Effects of pulsed electromagnetic fields and platelet rich plasma in preventing osteoclastogenesis in an in vitro model of osteolysis.
  Matilde Tschon, Francesca Veronesi, Deyanira Contartese, Maria Sartori, Lucia Martini, Fabrizio Vincenzi, Annalisa Ravani, Katia Varani, Milena Fini.
  Journal of Cellular Physiology. September 04, 2017
  Osteolysis is the main limiting cause for the survival of an orthopedic prosthesis and is accompanied by an enhancement in osteoclastogenesis and inflammation, due by wear debris formation. Unfortunately therapeutic treatments, besides revision surgery, are not available. The aim of the present study was to evaluate the effects of Pulsed Electro Magnetic Fields (PEMFs) and platelet rich plasma (PRP), alone or in combination, in an in vitro model of osteolysis. Rats peripheral blood mononuclear cells were cultured on Ultra High Molecular Weight Polyethylene particles and divided into four groups of treatments: (1) PEMF stimulation (12 hr/day, 2.5 mT, 75 Hz, 1.3 ms pulse duration); (2) 10% PRP; (3) combination of PEMFs, and PRP; (4) no treatment. Treatments were performed for 3 days and cell viability, osteoclast number, expression of genes related to osteoclastogenesis and inflammation and production of pro‐inflammatory cytokines were assessed up to 14 days. PEMF stimulation exerted best results because it increased cell viability at early time points and counteracted osteoclastogenesis at 14 days. On the contrary, PRP increased osteoclastogenesis and reduced cell viability in comparison to PEMFs alone. The combination of PEMFs and PRP increased cell viability over time and reduced osteoclastogenesis in comparison to PRP alone. However, these positive results did not exceed the level achieved by PEMF alone. At longer time points PEMF could not counteract osteoclastogenesis increased by PRP. Regarding inflammation, all treatments maintained the production of pro‐inflammatory cytokines at low level, although PRP increased the level of interleukin 1 beta. The aim of the present study was to evaluate the effects of Pulsed ElectroMagnetic Fields (PEMFs) and platelet rich plasma (PRP), alone or in combination, in an in vitro model of osteolysis.

  September 04, 2017   doi: 10.1002/jcp.26143   open full text
• Inhibition of ZL55 cell proliferation by ADP via PKC‐dependent signalling pathway.
  Antonella Muscella, Luca G. Cossa, Carla Vetrugno, Giovanna Antonaci, Santo Marsigliante.
  Journal of Cellular Physiology. September 04, 2017
  Extracellular nucleotides can regulate cell proliferation in both normal and tumorigenic tissues. Here, we studied how extracellular nucleotides regulate the proliferation of ZL55 cells, a mesothelioma‐derived cell line obtained from bioptic samples of asbestos‐exposed patients. ADP and 2‐MeS‐ADP inhibited ZL55 cell proliferation, whereas ATP, UTP, and UDP were inactive. The nucleotide potency profile and the blockade of the ADP‐mediated inhibitory effect by the phospholipase C inhibitor U‐73122 suggest that P2Y1 receptor controls ZL55 cell proliferation. The activation of P2Y1 receptor by ADP leads to activation of intracellular transduction pathways involving [Ca2+]i, PKC‐δ/PKC‐α, and MAPKs, ERK1/2 and JNK1/2. Cell treatment with ADP or 2‐MeS‐ADP also provokes the activation of p53, causing an accumulation of the G1 cyclin‐dependent kinase inhibitors p21WAF1 and p27Kip. Inhibition of ZL55 cell proliferation by ADP was completely reversed by inhibiting MEK1/2, or JNK1/2, or PKC‐δ, and PKC‐α. Through the inhibition of ADP‐activated transductional kinases it was found that PKC‐δ was responsible for JNK1/2 activation. JNK1/2 has a role in transcriptional up‐regulation of p53, p21WAF1/CIP1, and p27kip1. Conversely, the ADP‐activated PKC‐α provoked ERK1/2 phosphorylation. ERK1/2 increased p53 stabilization, required to G1 arrest of ZL55 cells. Concluding, the importance of the study is twofold: first, results shed light on the mechanism of cell cycle inhibition by ADP; second, results suggest that extracellular ADP may inhibit mesothelioma progression. In human mesothelioma ZL55 cells, activation of P2Y1 receptor by ADP leads to activation of intracellular transduction pathways involving [Ca2+]i, PKC‐δ/PKC‐α, and MAPKs ERK1/2 and JNK1/2. Cell treatment with ADP or 2‐MeS‐ADP also provokes the activation of p53, causing an accumulation of the G1 cyclin‐dependent kinase inhibitors p21WAF1 and p27Kip. ADP‐activated PKC‐δ was responsible of JNK1/2 activation that has a role in transcriptional up‐regulation of p53, p21WAF1, and p27kip1. Conversely, the ADP‐activated PKC‐α provoked ERK1/2 phosphorylation that increased p53 stabilization, required to G1 arrest of ZL55 cells. Concluding, the importance of the study is twofold: first, results shed light on the mechanism of cell cycle inhibition by ADP; second, results suggest that extracellular ADP may inhibit mesothelioma progression.

  September 04, 2017   doi: 10.1002/jcp.26128   open full text
• MicroRNAs in retinoblastoma: Potential diagnostic and therapeutic biomarkers.
  Khodayar Golabchi, Rahim Soleimani‐Jelodar, Nazila Aghadoost, Fatemeh Momeni, Abdullah Moridikia, Javid Sadri Nahand, Aria Masoudifar, Hasan Razmjoo, Hamed Mirzaei.
  Journal of Cellular Physiology. August 30, 2017
  Retinoblastoma (Rb) is known as one of important childhood malignancies which due to inactivation of the RB gene (tumor suppressor gene in various patients). The early detection of Rb could provide better treatment for Rb patients. Imaging techniques (e.g., MRI and CT) are known as one of effective diagnosis approaches for detection of patients with Rb. It has been shown that utilization of imaging techniques is associated with some limitations. Hence, identification of new diagnosis approaches might provide a better treatment for Rb patients. Identification of new biomarkers could contribute to better understanding of pathogenesis events involved in Rb and provide new insights into design better treatment approaches for these patients. Among the various biomarkers, microRNAs (miRNAs) have emerged as attractive tools for Rb detection. miRNAs are one classes of small non‐coding RNAs which could anticipate in a variety of biological process via targeting sequence of cellular and molecular pathways. Deregulations of these molecules are associated with cancerous condition. Multiple lines of evidence indicated that deregulation of various miRNAs involved in various stages of Rb. Here, we summarized a variety of tissue‐specific and circulating miRNAs involved in Rb pathogenesis which could be used as diagnostic, prognostic, and therapeutic biomarkers in Rb patients.

  August 30, 2017   doi: 10.1002/jcp.26070   open full text
• Endogenous bioelectric currents promote differentiation of the mammalian lens.
  Lin Cao, Jie Liu, Jin Pu, J. Martin Collinson, John V. Forrester, Colin D. McCaig.
  Journal of Cellular Physiology. August 30, 2017
  The functional roles of bioelectrical signals (ES) created by the flow of specific ions at the mammalian lens equator are poorly understood. We detected that mature, denucleated lens fibers expressed high levels of the α1 and β1 subunits of Na+/K+‐ATPase (ATP1A1 and ATP1B1 of the sodium pump) and had a hyperpolarized membrane potential difference (Vmem). In contrast, differentiating, nucleated lens fiber cells had little ATP1A1 and ATP1B1 and a depolarized Vmem. Mimicking the natural equatorial ES with an applied electrical field (EF) induced a striking reorientation of lens epithelial cells to lie perpendicular to the direction of the EF. An EF also promoted the expression of β‐crystallin, aquaporin‐0 (AQP0) and the Beaded Filament Structural Protein 2 (BFSP2) in lens epithelial cells (LECs), all of which are hallmarks of differentiation. In addition, applied EF activated the AKT and CDC2 and inhibition of AKT reduced the activation of CDC2. Our results indicate that the endogenous bioelectrical signal at the lens equator promotes differentiation of LECs into denucleated lens fiber cells via depolarization of Vmem. Development of methods and devices of EF application or amplification in vivo may supply a novel treatment for lens diseases and even promote regeneration of a complete new lens following cataract surgery. Mature, denucleated lens fibers expressed high levels of the α1 and β1 subunits of Na+/K+‐ATPase In contrast, differentiating, nucleated lens fiber cells had little ATP1A1 and ATP1B1. An applied electrical field (EF) induced a striking reorientation of lens epithelial cells to lie perpendicular to the direction of the EF. The applied EF activated the AKT and CDC2 and inhibition of AKT reduced the activation of CDC2.

  August 30, 2017   doi: 10.1002/jcp.26074   open full text
• Mesenchymal stem cells: A new platform for targeting suicide genes in cancer.
  Rana Moradian Tehrani, Javad Verdi, Mahdi Noureddini, Rasoul Salehi, Reza Salarinia, Meysam Mosalaei, Miganosh Simonian, Behrang Alani, Moosa Rahimi Ghiasi, Mahmoud Reza Jaafari, Hamed Reza Mirzaei, Hamed Mirzaei.
  Journal of Cellular Physiology. August 30, 2017
  One of the important strategies for the treatment of cancer is gene therapy which has the potential to exclusively eradicate malignant cells, without any damage to the normal tissues. Gene‐directed enzyme prodrug therapy (GDEPT) is a two‐step gene therapy approach, where a suicide gene is directed to tumor cells. The gene encodes an enzyme that expressed intracellularly where it is able to convert a prodrug into cytotoxic metabolites. Various delivery systems have been developed to achieve the appropriate levels of tumor restricted expression of chemotherapeutic drugs. Nowadays, mesenchymal stem cells (MSCs) have been drawing great attention as cellular vehicles for gene delivery systems. Inherent characteristics of MSCs make them particularly attractive gene therapy tools in cell therapy. They have been used largely for their remarkable homing property toward tumor sites and availability from many different adult tissues and show anti‐inflammatory actions in some cases. They do not stimulate proliferative responses of lymphocytes, suggests that MSCs have low immunogenicity and could avoid immune rejection. This review summarizes the current state of knowledge about genetically modified MSCs that enable to co‐transduce a variety of therapeutic agents including suicide genes (i.e., cytosine deaminase, thymidine kinase) in order to exert potent anti‐carcinogenesis against various tumors growth. Moreover, we highlighted the role of exosomes released from MSCs as new therapeutic platform for targeting various therapeutic agents.

  August 30, 2017   doi: 10.1002/jcp.26094   open full text
• Maresin 1 inhibits TNF‐alpha‐induced lipolysis and autophagy in 3T3‐L1 adipocytes.
  Laura M Laiglesia, Silvia Lorente‐Cebrián, Miguel López‐Yoldi, Raquel Lanas, Neira Sáinz, Jose Alfredo Martínez, Maria J. Moreno‐Aliaga.
  Journal of Cellular Physiology. August 30, 2017
  Obesity is associated with high levels of pro‐inflammatory cytokines such as tumor necrosis factor‐alpha (TNF‐α), which promotes inflammation in adipose tissue. The omega‐3 PUFAs, and their derived lipid mediators, such as Maresin 1 (MaR1) have anti‐inflammatory effects on adipose tissue. This study aimed to analyze if MaR1 may counteract alterations induced by TNF‐α on lipolysis and autophagy in mature 3T3‐L1 adipocytes. Our data revealed that MaR1 (1–100 nM) inhibited the TNF‐α‐induced glycerol release after 48 hr, which may be related to MaR1 ability of preventing the decrease in lipid droplet‐coating protein perilipin and G0/G1 Switch 2 protein expression. MaR1 also reversed the decrease in total hormone sensitive lipase (total HSL), and the ratio of phosphoHSL at Ser‐565/total HSL, while preventing the increased ratio of phosphoHSL at Ser‐660/total HSL and phosphorylation of extracellular signal‐regulated kinase 1/2 induced by TNF‐α. Moreover, MaR1 counteracted the cytokine‐induced decrease of p62 protein, a key autophagy indicator, and also prevented the induction of LC3II/LC3I, an important autophagosome formation marker. Current data suggest that MaR1 may ameliorate TNF‐α‐induced alterations on lipolysis and autophagy in adipocytes. This may also contribute to the beneficial actions of MaR1 on adipose tissue and insulin sensitivity in obesity. MaR1 counteracts the changes induced by TNF‐α on some of the main lipases and lipid droplets proteins controlling lipolysis (perilipin, HSL, and G0S2) as well as on the proteins regulating autophagy (p62 and LC3). These observations suggest that MaR1 may represent a promising therapeutic agent to counteract the alterations induced by inflammation in adipose tissue.

  August 30, 2017   doi: 10.1002/jcp.26096   open full text
• Tannic acid attenuates TGF‐β1‐induced epithelial‐to‐mesenchymal transition by effectively intervening TGF‐β signaling in lung epithelial cells.
  Dhamotharan Pattarayan, Ayyanar Sivanantham, Venkateshwaran Krishnaswami, Lakshmanan Loganathan, Rajaguru Palanichamy, Subramanian Natesan, Karthikeyan Muthusamy, Subbiah Rajasekaran.
  Journal of Cellular Physiology. August 30, 2017
  Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and an irreversible lung disorder characterized by the accumulation of fibroblasts and myofibroblasts in the extracellular matrix. The transforming growth factor‐β1 (TGF‐β1)‐induced epithelial‐to‐mesenchymal transition (EMT) is thought to be one of the possible sources for a substantial increase in the number of fibroblasts/myofibroblasts in IPF lungs. Tannic acid (TA), a natural dietary polyphenolic compound has been shown to possess diverse pharmacological effects. However, whether TA can inhibit TGF‐β1‐mediated EMT in lung epithelial cells remains enigmatic. Both the human adenocarcinomic alveolar epithelial (A549) and normal bronchial epithelial (BEAS‐2B) cells were treated with TGF‐β1 with or without TA. Results showed that TA addition, markedly inhibited TGF‐β1‐induced EMT as assessed by reduced expression of N‐cadherin, type‐1‐collagen, fibronectin, and vimentin. Furthermore, TA inhibited TGF‐β1‐induced cell proliferation through inducing cell cycle arrest at G0/G1 phase. TGF‐β1‐induced increase in the phosphorylation of Smad (Smad2 and 3), Akt as well as that of mitogen activated protein kinase (ERK1/2, JNK1/2, and p38) mediators was effectively inhibited by TA. On the other hand, TA reduced the TGF‐β1‐induced increase in TGF‐β receptors expression. Using molecular docking approach, FTIR, HPLC and Western blot analyses, we further identified the direct binding of TA to TGF‐β1. Finally, we conclude that TA might directly interact with TGF‐β1, thereby repressing TGF‐β signaling and subsequent EMT process in lung epithelial cells. Further animal studies are needed to clarify its potential therapeutic benefit in pulmonary fibrosis. Tannic acid suppresses TGF‐β1‐induced EMT.

  August 30, 2017   doi: 10.1002/jcp.26127   open full text
• Lipid composition of membrane microdomains isolated detergent‐free from PUFA supplemented RAW264.7 macrophages.
  Christine Hellwing, Feven Tigistu‐Sahle, Herbert Fuhrmann, Reijo Käkelä, Julia Schumann.
  Journal of Cellular Physiology. August 30, 2017
  Profound alterations in the lipid profile of raft and non‐raft plasma membrane microdomains were found when RAW264.7 macrophages were supplemented with polyunsaturated fatty acids (PUFAs) in physiologically relevant concentrations. For the first time lipids in the detergent‐free isolated membrane domains of phagocytic immune cells were characterized by mass spectrometry. The extent of remodeling of the membrane lipids differed with different n3 and n6 PUFA supplements. The mildest effects were detected for α‐linolenic acid (LNA) and linoleic acid (LA), the C18 precursors of the n3 and n6 families, respectively. When the effects of highly unsaturated PUFAs were compared, eicosapentaenoic acid (EPA) caused more extensive restructuring of membrane lipids than docosahexaenoic acid (DHA) or arachidonic acid (AA). The supplements altered the lipid species composition of both the raft and non‐raft membrane fractions. The rafts containing elevated proportions of highly unsaturated lipid species may relocate sterically incompatible lipids and proteins originally belonging to this microdomain. Such effect was evident for sphingomyelin, which favored non‐rafts instead of rafts after EPA supplementation. The current work suggests that the different functional consequences found previously when supplementing macrophages with either EPA or DHA have their origin in the different effects of these PUFAs on membrane architecture. Profound alterations in the lipid profile of raft and non‐raft plasma membrane microdomains were found when RAW264.7 macrophages were supplemented with polyunsaturated fatty acids (PUFAs) in physiologically relevant concentrations. The supplements altered the lipid species composition of both the raft and non‐raft membrane fractions. Furthermore, the current work suggests that the different functional consequences found previously when supplementing macrophages with either EPA or DHA have their origin in the different effects of these PUFAs on membrane architecture.

  August 30, 2017   doi: 10.1002/jcp.26138   open full text
• Annexin A2 positively regulates milk synthesis and proliferation of bovine mammary epithelial cells through the mTOR signaling pathway.
  Minghui Zhang, Dongying Chen, Zhen Zhen, Jinxia Ao, Xiaohan Yuan, Xuejun Gao.
  Journal of Cellular Physiology. August 30, 2017
  Annexin A2 (AnxA2) has been shown to play multiple roles in growth, development, and metabolism, but the functions of AnxA2 and the signaling pathways associated with AnxA2 are still not fully understood. In this study, we aim to reveal whether and how AnxA2 could be involved in milk synthesis and proliferation of bovine mammary epithelial cells (BMECs). Using gene function study approaches, we found that AnxA2 positively regulates PIP3 level, phosphorylation of mTOR, and protein levels of SREBP‐1c and Cyclin D1 leading to milk synthesis and cell proliferation. We further observed that both AnxA2‐36 kD phosphorylated form and AnxA2‐33 kD protein could be induced from AnxA2‐36 kD protein in BMECs under methionine, leucine, estrogen or prolactin stimulation. These above results strongly demonstrate that AnxA2 functions as a critical regulator for amino acid or hormone‐induced milk synthesis and cell proliferation via the PI3K‐mTOR‐SREBP‐1c/Cyclin D1 signaling pathway. Annexin A2 is phosphorylated and further processed into a truncated form in response to Met, Leu, E and PRL stimulation, and positively regulates milk synthesis and proliferation of bovine mammary epithelial cells through the mTOR signaling pathway

  August 30, 2017   doi: 10.1002/jcp.26123   open full text
• Basal progenitor cells bridge the development, malignant cancers, and multiple diseases of esophagus.
  Baoshun Lin, Fuan Xie, Zhangwu Xiao, Xiaoqian Hong, Liming Tian, Kuancan Liu.
  Journal of Cellular Physiology. August 30, 2017
  The esophagus is a pivotal organ originating from anterior foregut that links the mouth and stomach. Moreover, its development involves precise regulation of multiple signal molecules and signal transduction pathways. After abnormal regulation of these molecules in the basal cells of the esophagus occurs, multiple diseases, including esophageal atresia with or without tracheoesophageal fistula, Barrett esophagus, gastroesophageal reflux, and eosinophilic esophagitis, will take place as a result. Furthermore, expression changes of signal molecules or signal pathways in basal cells and the microenvironment around basal cells both can initiate the switch of malignant transformation. In this review, we highlight the molecular events underlying the transition of normal development to multiple esophageal diseases. Additionally, the animal models of esophageal development and related diseases, challenges, and strategies are extensively discussed. After abnormal regulation of these molecules in the basal cells of the esophagus occurs, multiple diseases, including esophageal atresia with or without tracheoesophageal fistula, Barrett esophagus, gastroesophageal reflux, and eosinophilic esophagitis, will take place as a result. Furthermore, expression changes of signal molecules or signal pathways in basal cells and the microenvironment around basal cells both can initiate the switch of malignant transformation.

  August 30, 2017   doi: 10.1002/jcp.26136   open full text
• Crosstalk of ER stress‐mediated autophagy and ER‐phagy: Involvement of UPR and the core autophagy machinery.
  Shuling Song, Jin Tan, Yuyang Miao, Qiang Zhang.
  Journal of Cellular Physiology. August 30, 2017
  Endoplasmic reticulum (ER) stress, a common cellular stress response, is closely related to the activation of autophagy that is an important and evolutionarily conserved mechanism for maintaining cellular homeostasis. Autophagy induced by ER stress mainly includes the ER stress‐mediated autophagy and ER‐phagy. The ER stress‐mediated autophagy is characterized by the generation of autophagosomes that include worn‐out proteins, protein aggregates, and damaged organelles. While the autophagosomes of ER‐phagy selectively include ER membranes, and the double membranes also derive, at least in part, from the ER. The signaling pathways of IRE1α, PERK, ATF6, and Ca2+ are necessary for the activation of ER stress‐mediated autophagy, while the receptor‐mediated selective ER‐phagy degrades the ER is Atg40/FAM134B. The ER stress‐mediated autophagy and ER‐phagy not only have differences, but also have connections. The activation of ER‐phagy requires the core autophagy machinery, and the ER‐phagy may be a branch of ER stress‐mediated autophagy that selectively targets the ER. However, the determined factors that control the changeover switch between ER stress‐mediated autophagy and ER‐phagy are largely obscure, which may be associated with the type of cells and the extent of stimulation. This review summarized the crosstalk between ER stress‐mediated autophagy and ER‐phagy and their signaling networks. Additionally, we discussed the possible factors that influence the type of autophagy induced by ER stress. Under ER stress condition, the ER stress‐mediated autophagy and ER‐phagy can be activated, and both involved in UPR and the core autophagy machinery. However, the determined factors that control the changeover switch between ER stress‐mediated autophagy and ER‐phagy, are unclear. And we speculate that the type of cells and the extent of stimulation may be associated with the type of autophagy induced by ER stress.

  August 30, 2017   doi: 10.1002/jcp.26137   open full text
• EPA blocks TNF‐α‐induced inhibition of sugar uptake in Caco‐2 cells via GPR120 and AMPK.
  Rosa Castilla‐Madrigal, Jaione Barrenetxe, María J. Moreno‐Aliaga, María Pilar Lostao.
  Journal of Cellular Physiology. August 30, 2017
  The aim of the present work was to investigate in Caco‐2 cells whether eicosapentaenoic acid (EPA), an omega‐3 polyunsaturated fatty acid, could block the inhibitory effect of tumor necrosis factor‐α (TNF‐α) on sugar transport, and identify the intracellular signaling pathways involved. After pre‐incubation of the Caco‐2 cells with TNF‐α and EPA for 1 hr, EPA prevented the inhibitory effect of the cytokine on α‐methyl‐d‐glucose (αMG) uptake (15 min) and on SGLT1 expression at the brush border membrane, measured by Western blot. The ERK1/2 inhibitor PD98059 and the AMPK activator AICAR also prevented the inhibitory effect of TNF‐α on both αMG uptake and SGLT1 expression. Interestingly, the AMPK inhibitor, Compound C, abolished the ability of EPA to prevent TNF‐α‐induced reduction of sugar uptake and transporter expression. The GPR120 antagonist, AH7614, also blocked the preventive effect of EPA on TNF‐α‐induced decrease of αMG uptake and AMPK phosphorylation. In summary, TNF‐α inhibits αMG uptake by decreasing SGLT1 expression in the brush border membrane through the activation of ERK1/2 pathway. EPA prevents the inhibitory effect of TNF‐α through the involvement of GPR120 and AMPK activation. TNF‐α decreases SGLT1 expression in the plasma membrane by activation of ERK and inhibition of AMPK pathways. EPA prevents TNF‐α decrease of sugar uptake by activating AMPK pathways. EPA blocks TNF‐α effect on sugar uptake through GPR120.

  August 30, 2017   doi: 10.1002/jcp.26115   open full text
• β‐asarone inhibited cell growth and promoted autophagy via P53/Bcl‐2/Bclin‐1 and P53/AMPK/mTOR pathways in Human Glioma U251 cells.
  Nanbu Wang, Qinxin Zhang, Laiyu Luo, Baile Ning, Yongqi Fang.
  Journal of Cellular Physiology. August 30, 2017
  Glioma is the most common type of primary brain tumor and has an undesirable prognosis. Autophagy plays an important role in cancer therapy, but it is effect is still not definite. P53 is an important tumor suppressor gene and protein that is closely to autophagy. Our aim was to study the effect of β‐asarone on inhibiting cell proliferation in human glioma U251 cells and to detect the effect of the inhibition on autophagy through the P53 signal pathway. For cell growth, the cells were divided into four groups: the model, β‐asarone, temozolomide (TMZ), and co‐administration groups. For cell autoghapy and the P53 pathway, the cells were divided into six groups: the model, β‐asarone, 3MA, Rapa, Pifithrin‐µ, and NSC groups. The counting Kit‐8 assay and flow cytometry (FCM) were then used to measure the cell proliferation and cycle. Electron microscopy was used to observe autophagosome formation. Cell immunohistochemistry/‐immunofluorescence, FCM and Western blot (WB) were used to examine the expression of Beclin‐1 and P53. The levels of P53 and GAPDH mRNA were detected by RT‐PCR. Using WB, we determined autophagy‐related proteins Beclin‐1, LC3‐II/I, and P62 and those of the P53 pathway‐related proteins P53, Bcl‐2, mTOR, P‐mTOR, AMPK, P‐AMPK, and GAPDH. We got the results that β‐asarone changed the cellular morphology, inhibited cell proliferation, and enhanced the expression of P53, LC3‐II/I, Beclin‐1, AMPK, and pAMPK while inhibiting the expression of P62, Bcl‐2, mTOR, and pmTOR. All the data suggested that β‐asarone could reduce the cell proliferation and promote autophagy possible via the P53 pathway in U251 cells. β‐asarone inhibits the growth of glioma U251 cells. β‐asarone promotes autophagy of U251 cells. The autophagy process possible be promoted through the P53/Bcl‐2/Bclin‐1 and P53/AMPK/mTOR signal pathway.

  August 30, 2017   doi: 10.1002/jcp.26118   open full text
• TLR4 and NFκB signaling is critical for taxol resistance in ovarian carcinoma cells.
  Nian‐Kang Sun, Shang‐Lang Huang, Ting‐Chang Chang, Chuck C.‐K. Chao.
  Journal of Cellular Physiology. August 30, 2017
  We report here that toll‐like receptor 4 (TLR4) and ABCB1 are upregulated in SKOV3 ovarian carcinoma cells that acquired resistance to the anticancer drug taxol. Silencing of TLR4 using short‐hairpin RNA sensitized taxol‐resistant SKOV3 cells to taxol (4.6 fold), whereas ectopic expression of TLR4 in parental, taxol‐sensitive SKOV3 cells or TLR4‐null HEK293 cells induced taxol resistance (∼2 fold). A sub‐lethal dose of taxol induced ABCB1 protein expression in taxol‐resistant SKOV3 cells. Inactivation of TLR4 using chemical inhibitors (CLI‐095 and AO‐I) downregulated ABCB1 protein expression and enhanced the cytotoxic activity of taxol in taxol‐resistant SKOV3 cells. While the sensitization effect of TLR4 inactivation was also detected in TOV21G ovarian cancer cells, which express moderate level of TLR4, ectopic expression of ABCB1 prevented the sensitization effect in these cells. Notably, the NFκB pathway was significantly activated by taxol, and inhibition of this pathway suppressed TLR4‐regulated ABCB1 expression. Furthermore, taxol‐induced NFκB signaling was reduced following TLR4 silencing in taxol‐resistant SKOV3 cells. Consistent with these results, ectopic expression of TLR4 in taxol‐sensitive SKOV3 cells enhanced ABCB1 expression and conferred resistance to taxol. The protective effect of exogenous TLR4 expression against taxol was reduced by treatment with NFκB inhibitor in these cells. These results demonstrate that taxol activates the TLR4‐NFκB pathway which in turn induces ABCB1 gene expression. This cellular pathway thus represents a novel target to limit resistance to taxol in ovarian cancer cells. The taxol activates the TLR4‐NFkB pathway which in turn induces ABCB1 gene expression. This cellular pathway thus represents a novel target to limit resistance to taxol in ovarian cancer cells.

  August 30, 2017   doi: 10.1002/jcp.26125   open full text
• Hypomethylation‐mediated H19 overexpression increases the risk of disease evolution through the association with BCR‐ABL transcript in chronic myeloid leukemia.
  Jing‐dong Zhou, Jiang Lin, Ting‐juan Zhang, Ji‐chun Ma, Xi‐xi Li, Xiang‐mei Wen, Hong Guo, Zi‐jun Xu, Zhao‐qun Deng, Wei Zhang, Jun Qian.
  Journal of Cellular Physiology. August 30, 2017
  Previous study has revealed that H19 expression is required for efficient tumor growth induced by BCR‐ABL in chronic myeloid leukemia (CML). Herein, we further determined H19 expression and its clinical implication in patients with CML. H19 expression and methylation were detected by real‐time quantitative PCR and real‐time quantitative methylation‐specific PCR, and then clinical implication of H19 expression was further analyzed. H19 expression was significantly up‐regulated in CML patients (p < 0.001). H19 expression with an area under receiver operating characteristic curve value of 0.824 might serve as a promising biomarker in distinguishing CML patients from controls. The patients with high H19 expression had a tendency of higher white blood cells and BCR‐ABL transcript than those with low H19 expression. H19 overexpression occurred with the higher frequency in blast crisis stage (11/11, 100%), lower in accelerated phase (3/5, 60%), and chronic phase (42/62, 66%) stages. Moreover, paired patients during disease progression with increased BCR‐ABL transcript also showed a significant upregulation of H19 expression. Meanwhile, H19 expression was decreased in follow‐up patients who achieved complete molecular remission after tyrosine kinase inhibitors‐based therapy. Epigenetic studies showed that H19 differentially methylated region/imprinting control region (DMR/ICR) was hypomethylated and associated with H19 expression in CML patients. Moreover, demethylation of H19 DMR/ICR reactivated H19 expression in K562 cells. Collectively, H19 overexpression, a frequent event in CML, was associated with higher BCR‐ABL transcript involving in disease progression. Moreover, H19 DMR/ICR hypomethylation in CML may be one of the mechanisms mediating H19 overexpression. H19 overexpression, a frequent event in CML, was associated with higher BCR‐ABL transcript involving in disease progression. H19 DMR/ICR hypomethylation in CML may be one of the mechanisms mediating H19 overexpression.

  August 30, 2017   doi: 10.1002/jcp.26119   open full text
• 12‐O‐tetradecanoylphorbol‐13‐acetate and EZH2 inhibition: A novel approach for promoting myogenic differentiation in embryonal rhabdomyosarcoma cells.
  Irene Marchesi, Luca Sanna, Milena Fais, Francesco P. Fiorentino, Antonio Giordano, Luigi Bagella.
  Journal of Cellular Physiology. August 30, 2017
  Rhabdomyosarcoma (RMS) is a soft tissue sarcoma that arises from muscle precursors affecting predominately children and young adults. It can be divided into two main classes: embryonal (eRMS) and alveolar rhabodomyosarcomas (aRMS). Despite the expression of early muscle specific genes, RMS cells fail to complete myogenesis even in differentiation conditions. We previously demonstrated that Enhancer Zeste of Homolog 2 (EZH2), the catalytic subunits of PRC2 complex, contributes to inhibit muscle differentiation in eRMS and its down‐regulation causes a partial recovery of myogenesis. 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) is a molecule able to induce differentiation in eRMS with a mechanism that involves the protein kinase C (PKC). In this paper we report that treatment with TPA reduces the expression of EZH2 without affecting levels of H3K27me3. The combination of TPA with GSK126, an inhibitor of the catalytic activity of EZH2, has a synergic effect on the induction of muscle differentiation in RD rhabdomyosarcoma cells, suggesting a new therapeutic combinatory approach for RMS treatment. Rhabdomyosarcoma (RMS) is a soft tissue sarcoma that arises from muscle precursors. We previously demonstrated that down‐regulation of EZH2 in RMS cells causes a partial recovery of myogenesis. We report that the combination of TPA with GSK126 scores a synergic effect on the induction of muscle differentiation, suggesting a new therapeutic combinatory approach for RMS treatment

  August 30, 2017   doi: 10.1002/jcp.26107   open full text
• Aquaporin1 and 3 modification as a result of chondrogenic differentiation of human mesenchymal stem cell.
  Adriana C. E. Graziano, Rosanna Avola, Giovanna Pannuzzo, Venera Cardile.
  Journal of Cellular Physiology. August 28, 2017
  Chondrocytes are cells of articular cartilage particularly sensitive to water transport and ionic and osmotic changes from extracellular environment and responsible for the production of the synovial fluid. Aquaporins (AQPs) are a family of water and small solute transport channel proteins identified in several tissues, involved in physiological pathways and in manifold human diseases. In a recent period, AQP1 and 3 seem to have a role in metabolic water regulation in articular cartilage of load bearing joints. The aim of this study was to examine the levels of AQP1 and 3 during the chondrogenic differentiation of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT). For the determination of chondrogenic markers and AQPs levels, glycosaminoglycans (GAGs) quantification, immunocytochemistry, RT‐PCR, and Western blot were used after 0, 7, 14, 21, and 28 days from the start of differentiation. At 21 days, chondrocytes derived from AT‐MSCs were able to produce augmented content of GAGs and significant quantity of SOX‐9, lubricin, aggrecan, and collagen type II, suggesting hyaline cartilage formation, in combination with an increase of AQP3 and AQP1. However, while AQP1 level decreased after 21 days; AQP3 reached higher values at 28 days. The expression of AQP1 and 3 is a manifestation of physiological adaptation of functionally mature chondrocytes able to respond to the change of their internal environment influenced by extracellular matrix. The alteration or loss of expression of AQP1 and 3 could contribute to destruction of chondrocytes and to development of cartilage damage. We examined the levels of AQP1 and 3 during the chondrogenic differentiation of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT). At 21 days, chondrocytes derived from AT‐MSCs were able to produce augmented content of GAGs and significant quantity of SOX‐9, lubricin, aggrecan and collagen type II, suggesting hyaline cartilage formation, In parallel, the same cells increased AQP3 and AQP1. AQP1 and AQP3 can be functionally involved in homeostasis and chondrogenic differentiation of AT‐MSCs.

  August 28, 2017   doi: 10.1002/jcp.26100   open full text
• Structure‐based release analysis of the JC virus agnoprotein regions: A role for the hydrophilic surface of the major alpha helix domain in release.
  A. Sami Saribas, Martyn K. White, Mahmut Safak.
  Journal of Cellular Physiology. August 28, 2017
  Agnoprotein (Agno) is an important regulatory protein of JC virus (JCV), BK virus (BKV) and simian virus 40 (SV40) and these viruses are unable to replicate efficiently in the absence of this protein. Recent 3D‐NMR structural data revealed that Agno contains two alpha‐helices (a minor and a major) while the rest of the protein adopts an unstructured conformation (Coric et al., 2017, J Cell Biochem). Previously, release of the JCV Agno from the Agno‐positive cells was reported. Here, we have further mapped the regions of Agno responsible for its release by a structure‐based systematic mutagenesis approach. Results revealed that amino acid residues (Lys22, Lys23, Phe31, Glu34, and Asp38) located either on or adjacent to the hydrophilic surface of the major alpha‐helix domain of Agno play critical roles in release. Additionally, Agno was shown to strongly interact with unidentified components of the cell surface when cells are treated with Agno, suggesting additional novel roles for Agno during the viral infection cycle. JC virus agnoprotein plays critical regulatory roles during the viral replication cycle. Previously, the release of agnoprotein from agnoprotein‐positive cells was reported. In this work, we have further mapped the region(s) responsible for its release. Data showed that the specific amino acid residues located on the hydrophilic surface (designated as release surface) of the major alpha helix domain of agnoprotein, including Lys22, Lys23, Phe31, Glu34, and Asp38 play important roles in this process. Additionally, agnoprotein was found to strongly interact with the unidentified components of the cell surface, suggesting additional roles for it during the viral replication cycle.

  August 28, 2017   doi: 10.1002/jcp.26106   open full text
• miRNAs and ovarian cancer: An overview.
  Bornali Deb, Arif Uddin, Supriyo Chakraborty.
  Journal of Cellular Physiology. August 25, 2017
  Ovarian cancer (OC) is the sixth most common cancer in women globally. However, even with the advances in detection and therapeutics it still represents the most dangerous gynecologic malignancy in women of the industrialized countries. The discovery of micro‐RNAs (miRNA), a small noncoding RNA molecule targeting multiple mRNAs and regulation of gene expression by triggering translation repression and/or RNA degradation, has revealed the existence of a new array for regulation of genes involved in cancer. This review summarizes the current knowledge regarding the role of miRNAs expression in OC. It also provides information about potential clinical relevance of circulating miRNAs for OC diagnosis, prognosis, and therapeutics. The identification of functional targets for miRNAs represents a major obstacle in our understanding of microRNA function in OC, but significant progress is being made. The better understanding of the role of microRNA expression in ovarian cancer may provide new array for the detection, diagnosis, and therapy of the OC. This review summarizes the current knowledge regarding the role of miRNAs expression in OC. It also provides information about potential clinical relevance of circulating miRNAs for OC diagnosis, prognosis, and therapeutics. The identification of functional targets for miRNAs represents a major obstacle in our understanding of microRNA function in OC, but significant progress is being made. The better understanding of the role of microRNA expression in ovarian cancer may provide new array for the detection, diagnosis, and therapy of the OC.

  August 25, 2017   doi: 10.1002/jcp.26095   open full text
• miR‐199a‐3p is involved in estrogen‐mediated autophagy through the IGF‐1/mTOR pathway in osteocyte‐like MLO‐Y4 cells.
  Jiayao Fu, Lingyu Hao, Yawen Tian, Yang Liu, Yijing Gu, Junhua Wu.
  Journal of Cellular Physiology. August 25, 2017
  To date, evidence indicates that estrogen partially modulates cellular processes through microRNAs. Autophagy is a catabolic process that is regulated by multiple factors and is associated with skeletal diseases. However, whether estrogen regulates osteocyte autophagy via microRNAs is largely unknown. In this study, we observed the up‐regulation of microRNA‐199a‐3p, a post‐transcriptional regulatory factor, in osteocytic areas in ovariectomized (OVX) mice. The mature forms of miR‐199a‐3p and pri‐miR‐199a were produced in response to estrogen signaling in osteocyte‐like MLO‐Y4 cells. Western blotting, autophagic flux detection, mRFP‐GFP‐LC3 fluorescence, and electron microscopy confirmed that miR‐199a‐3p induced autophagy in MLO‐Y4 cells, although cellular apoptosis was not affected. Additionally, we documented the ability of estrogen to mediate osteocyte autophagy. Based on our in vivo data, estrogen deficiency induced autophagy in osteocytes. Treatment of starved MLO‐Y4 cells with 17β‐estradiol suppressed the excess autophagy induced by starvation via activation of mammalian target of rapamycin (mTOR)‐related signaling cascades, while administration of rapamycin reversed the effects of 17β‐estradiol. Meanwhile, miR‐199a‐3p overexpression reversed 17β‐estradiol‐mediated regulation of autophagy in MLO‐Y4 cells. According to mechanistic studies, miR‐199a‐3p inhibited the mTOR pathway by directly binding to the 3′‐untranslated regions of insulin growth factor‐1 (IGF‐1) and mTOR. However, overexpression of miR‐199a‐3p inhibited IGF‐1 phosphorylation and mTOR‐related pathways. Knockdown of mTOR and IGF‐1 abolished estrogen signaling and restored LC3‐II expression through mTOR re‐activation, respectively. Thus, miR‐199a‐3p appears to be involved in the estrogen regulatory networks that mediate bone cell autophagy, potentially by targeting IGF‐1 and mTOR. miR‐199a‐3p is mediated by estrogen signaling in osteocyte both in vivo and in vitro. miR‐199a‐3p induces autophagy in MLO‐Y4 cells via directly binding to IGF‐1 and mTOR.

  August 25, 2017   doi: 10.1002/jcp.26101   open full text
• Mineral trioxide aggregate enhances the osteogenic capacity of periodontal ligament stem cells via NF‐κB and MAPK signaling pathways.
  Yanqiu Wang, Yixiang Zhou, Lin Jin, Xiyao Pang, Yadie Lu, Zilu Wang, Yan Yu, Jinhua Yu.
  Journal of Cellular Physiology. August 25, 2017
  Mineral trioxide aggregate (MTA), as a bioactive material, has a widespread application in clinical practice. To date, the effects of MTA on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs) remain unclear. hPDLSCs were isolated from human periodontal ligament tissues and cultured with MTA conditioned media. Cell counting kit‐8 (CCK‐8) assay was performed to assess the proliferation capacity of MTA‐treated hPDLSCs. Immunofluorescence assay, alkaline phosphatase (ALP) activity, alizarin red staining, real‐time RT‐PCR, and western blot analyses were used to investigate the odonto/osteogenic capacity of hPDLSCs as well as the involvement of NF‐κB and MAPK pathways. ALP activity assay revealed that 2 mg/ml was the optimal concentration for the induction of hPDLSCs by MTA. The protein expression of DSP, RUNX2, OCN, OSX, OPN, DMP1, ALP, and COL‐I in MTA‐treated hPDLSCs was significantly higher than those in control group (p < 0.01). When hPDLSCs were treated with the inhibitors of NF‐κB and MAPK pathways (U0126, SP600125, SB203580, and BMS345541), the effects of MTA on the differentiation of hPDLSCs were suppressed. Mechanistically, P65 was detected to transfer from cytoplasm to nuclei, as indicated by western blot and immunofluorescence assay. Moreover, MAPK‐related proteins and its downstream transcription factors were also upregulated in MTA‐treated hPDLSCs. Together, mineral trioxide aggregate can promote the odonto/osteogenic capacity of hPDLSCs via activating the NF‐κB and MAPK pathways. In this study, MTA was used to investigate its influence on the proliferation and differentiation of hPDLSCs. 2 mg/ml MTA promoted the osteo/odontogenic differentiation of hPDLSCs. Furthermore, NF‐κB and MAPK pathways were activated in MTA‐treated hPDLSCs, accompanying with the up‐regulation of downstream transcription factors.

  August 25, 2017   doi: 10.1002/jcp.26110   open full text
• Cyanidin suppresses autophagic activity regulating chondrocyte hypertrophic differentiation.
  Zhen Cao, Song Huang, Ce Dou, Qiang Xiang, Shiwu Dong.
  Journal of Cellular Physiology. August 25, 2017
  Cartilage is a kind of special connective tissue which does not contain neither blood vessels nor lymphatics and nerves. Therefore, the damage in cartilage is difficult to be repaired spontaneously. Constructing tissue engineered cartilage provides a new technique for cartilage repairing. Mesenchymal stem cells (MSCs) possess a unique capability of self‐renew and can differentiate into pre‐chondrocytes which are frequently applied as seed cells in tissue engineering. However, in regenerated cartilage the chondrocytes derived from MSCs can hardly maintain homeostasis and preferentially present hypertrophic like phenotype. We investigated the effects of cyanidin, a natural organic compound, on chondrogenic and subsequent hypertrophic differentiation of MSCs in order to seek approaches to inhibit chondrocyte hypertrophy. We evaluated the effects of cyanidin on expression of chondrogenic and hypertrophic marker genes through RT‐PCR, Western blot, alcian blue staining, and immunocytochemistry. The results showed that both chondrogenic related genes Sox9, Col2a1, and hypertrophic marker genes Runx2, Col10a1 were inhibited by cyanidin. In addition, we found that cyanidin promoted Nrf2 and p62 expression and suppressed LC3B expression during chondrogenic stage of MSCs. Meanwhile phosphorylation of IκBα and autophagosome related protein LC3B were inactivated by cyanidin during chondrocyte hypertrophic stage. Furthermore, rapamycin, an autophagy activator, abrogated the inhibitory effect of cyanidin on chondrogenic, and hypertrophic differentiation of MSCs. In conclusion, one potential mechanism of cyanidin, by which the chondrogenic and hypertrophic differentiation of MSCs were inhibited, was due to decreased autophagy activity. Our results indicated that cyanidin was a potential therapeutic agent for keeping mature chondrocyte functions. Cyanidin increased Nrf2 expression and inactivated autophagy to inhibit chondrogenic differentiation of MSCs. Besides, cyanidin inhibited NF‐κB signaling and suppressed autophagy to delay chondrocyte hypertrophic differentiation.

  August 25, 2017   doi: 10.1002/jcp.26105   open full text
• Simvastatin induces G1 arrest by up‐regulating GSK3β and down‐regulating CDK4/cyclin D1 and CDK2/cyclin E1 in human primary colorectal cancer cells.
  Ming‐Jenn Chen, An‐Ching Cheng, Ming‐Fen Lee, Yi‐Chiang Hsu.
  Journal of Cellular Physiology. August 18, 2017
  Simvastatin (SIM), a widely used cholesterol‐lowering drug, also exhibits tumor‐suppressive potentials in several types of malignancy. Colorectal cancer (CRC), the third most common malignant neoplasm, accounts for the second most leading cause of cancer‐related deaths worldwide. In the present study, we investigated the anticancer effects of SIM on CRC using primary cancer cells lines (CPs: CP1 to CP5) isolated from five Taiwanese colorectal cancer patients as a model for colorectal cancer. We treated all five CPs with SIM for 24 to 72 hours and observed the respective cell viability by an MTT assay. SIM increased DNA content of the G1 phase, but did not induce apoptosis/necrosis in CPs as shown by flow cytometry with propidium iodide (PI)/annexin V double staining and PI staining. The expression of G1 phase‐related proteins was analyzed by RT‐PCR and Western blotting. SIM suppressed cell growth and induced cell cycle G1‐arrest by suppressing the expression of CDK4/cyclin D1 and CDK2/cyclin E1, but elevating the expression of glycogen synthase kinase 3β in CPs. Our findings indicate that SIM may have antitumor activity in established colorectal cancer. This article is protected by copyright. All rights reserved

  August 18, 2017   doi: 10.1002/jcp.26156   open full text
• Tetrahydrocurcumin Ameliorates Homocysteine Mediated Mitochondrial Remodeling in Brain Endothelial Cells.
  Jonathan C. Vacek, Jyotirmaya Behera, Akash K George, Pradip K Kamat, Anuradha Kalani, Neetu Tyagi.
  Journal of Cellular Physiology. August 18, 2017
  Homocysteine (Hcy) causes endothelial dysfunction by inducing oxidative stress in most neurodegenerative disorders. This dysfunction is highly correlated with mitochondrial dynamics such as fusion and fission. However, there are no strategies to prevent Hcy induced mitochondrial remodeling. Tetrahydrocurcumin (THC) is an anti‐inflammatory and anti‐oxidant compound. We hypothesized that THC may ameliorates Hcy induced mitochondria remodeling in mouse brain endothelial cells (bEnd3) cells. bEnd3 cells were exposed to Hcy treatment in the presence or absence of THC. Cell viability and autophagic cell death were measured with MTT and MDC staining assay. Reactive oxygen species (ROS) production was determined using DCFH‐DA staining by confocal microscopy. Autophagy flux was assessed using a conventional GFP‐microtubule‐associated protein 1 light chain 3 (LC3) dot assay. Interaction of phagophore marker LC‐3 with mitochondrial receptor NIX was observed by confocal imaging. Mitochondrial fusion and fission were evaluated by western blot and RT‐PCR. Our results demonstrated that Hcy resulted in cell toxicity in a dose‐dependent manner and supplementation of THC prevented the detrimental effects of Hcy on cell survival. Furthermore, Hcy also upregulated of fission marker (DRP‐1), fusion markers (Mfn2) and autophagy marker (LC‐3). Finally, we observed that Hcy activated mitochondrial specific phagophore marker (LC‐3) was co‐localized with the mitochondrial receptor NIX, as viewed by confocal microscopy. Pretreatment of bEnd3 with THC (15µM) ameliorated Hcy induced oxidative damage, mitochondrial fission/fusion, and mitophagy. Our studies strongly suggest that THC has beneficial effects on mitochondrial remodeling and could be developed as a potential therapeutic agent against hyperhomocysteinemia (HHcy) induced mitochondrial dysfunction. This article is protected by copyright. All rights reserved

  August 18, 2017   doi: 10.1002/jcp.26145   open full text
• Chrysin Attenuates Progression of Ovarian Cancer Cells by Regulating Signaling Cascades and Mitochondrial Dysfunction.
  Whasun Lim, Soomin Ryu, Fuller W. Bazer, Sung‐Man Kim, Gwonhwa Song.
  Journal of Cellular Physiology. August 17, 2017
  Chrysin is mainly found in passion flowers, honey, and propolis acts as a potential therapeutic and preventive agent to inhibit proliferation and invasion of various human cancer cells. Although chrysin has anti‐carcinogenic effects in several cancers, little is known about its functional roles in ovarian cancer which shows poor prognosis and chemoresistance to traditional therapeutic agents. In the present study, we investigated functional roles of chrysin in progression of ovarian cancer cells using ES2 and OV90 (clear cell and serous carcinoma, respectively) cell lines. Results of the current study demonstrated that chrysin inhibited ovarian cancer cell proliferation and induced cell death by increasing reactive oxygen species (ROS) production and cytoplasmic Ca2+ levels as well as inducing loss of mitochondrial membrane potential (MMP). Moreover, chrysin activated mitogen‐activated protein kinase (MAPK) and phosphoinositide 3‐kinase (PI3K)/AKT pathways in ES2 and OV90 cells in concentration‐response experiments. Collectively, our results led us to propose that chrysin‐induced apoptotic events are mediated by the activation of PI3K and MAPK pathways in human ovarian cancer cells. This article is protected by copyright. All rights reserved

  August 17, 2017   doi: 10.1002/jcp.26150   open full text
• C‐C Motif Chemokine Ligand 2 Regulates LPS‐Induced Inflammation and ER Stress to Enhance Proliferation of Bovine Endometrial Epithelial Cells.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Sung‐Man Kim, Gwonhwa Song.
  Journal of Cellular Physiology. August 17, 2017
  Chemokines play an important role in regulating the complex immune system at the maternal‐fetal interface during pregnancy. Among various chemokines, C‐C motif chemokine ligand 2 (CCL2) plays a role in the recruitment of immune regulatory cells to implantation sites within the endometrium. In cattle, CCL2 is abundantly expressed in the uterine endometrium. However, its intracellular signaling has not been identified. In this study, we examined the effects of CCL2 on bovine endometrial (BEND) cell proliferation. CCL2 stimulated BEND cell proliferation by abundant expression of PCNA, accumulation of cells in the G2/M phase, and activation of the PI3K/AKT and MAPK signaling pathways. Moreover, CCL2 reduced endoplasmic reticulum stress and restored the inflammation‐induced reduction in BEND cell proliferation by regulating the unfolded protein response genes and cytokines. Collectively, these results demonstrated that CCL2 plays a pivotal role in reproductive tissues and may support maternal‐fetal interface to improve efficiency of pregnancy. This article is protected by copyright. All rights reserved

  August 17, 2017   doi: 10.1002/jcp.26151   open full text
• Artesunate inhibits RANKL‐induced osteoclastogenesis and bone resorption in vitro and prevents LPS‐induced bone loss in vivo.
  Cheng‐Ming Wei, Qian Liu, Fang‐Ming Song, Xi‐Xi Lin, Yi‐Ji Su, Jiake Xu, Lin Huang, Shao‐Hui Zong, Jin‐Min Zhao.
  Journal of Cellular Physiology. August 17, 2017
  Osteoclasts are multinuclear giant cells responsible for bone resorption in lytic bone diseases such as osteoporosis, arthritis, periodontitis, and bone tumors. Due to the severe side‐effects caused by the currently available drugs, a continuous search for novel bone‐protective therapies is essential. Artesunate (Art), the water‐soluble derivative of artemisinin has been investigated owing to its anti‐malarial properties. However, its effects in osteoclastogenesis have not yet been reported. In this study, Art was shown to inhibit the nuclear factor‐κB ligand (RANKL)‐induced osteoclastogenesis, the mRNA expression of osteoclastic‐specific genes, and resorption pit formation in a dose‐dependent manner in primary bone marrow‐derived macrophages cells (BMMs). Furthermore, Art markedly blocked the RANKL‐induced osteoclastogenesis by attenuating the degradation of IκB and phosphorylation of NF‐κB p65. Consistent with the in vitro results, Art inhibited lipopolysaccharide (LPS)‐induced bone resorption by suppressing the osteoclastogenesis. Together our data demonstrated that Art inhibits RANKL‐induced osteoclastogenesis by suppressing the NF‐κB signaling pathway and that it is a promising agent for the treatment of osteolytic diseases. Artesunate (Art), the water‐soluble derivative of artemisinin has been investigated owing to its anti‐malarial properties. However, its effects on osteoclastogenesis have not yet been reported. In this study, Art was shown to inhibit the nuclear factor‐kB ligand (RANKL)‐induced osteoclastogenesis and lipopolysaccharide (LPS)‐induced bone resorption.

  August 17, 2017   doi: 10.1002/jcp.25907   open full text
• Characterization and assessment of potential microRNAs involved in phosphate‐induced aortic calcification.
  Maya Fakhry, Najwa Skafi, Mohammad Fayyad‐Kazan, Firas Kobeissy, Eva Hamade, Saida Mebarek, Aida Habib, Nada Borghol, Asad Zeidan, David Magne, Hussein Fayyad‐Kazan, Bassam Badran.
  Journal of Cellular Physiology. August 04, 2017
  Medial artery calcification, a hallmark of type 2 diabetes mellitus and chronic kidney disease (CKD), is known as an independent risk factor for cardiovascular mortality and morbidity. Hyperphosphatemia associated with CKD is a strong stimulator of vascular calcification but the molecular mechanisms regulating this process remain not fully understood. We showed that calcification was induced after exposing Sprague‐Dawley rat aortic explants to high inorganic phosphate level (Pi, 6 mM) as examined by Alizarin red and Von Kossa staining. This calcification was associated with high Tissue‐Nonspecific Alkaline Phosphatase (TNAP) activity, vascular smooth muscle cells de‐differentiation, manifested by downregulation of smooth muscle 22 alpha (SM22α) protein expression which was assessed by immunoblot analysis, immunofluorescence, and trans‐differentiation into osteo‐chondrocyte‐like cells revealed by upregulation of Runt related transcription factor 2 (Runx2), TNAP, osteocalcin, and osteopontin mRNA levels which were determined by quantitative real‐time PCR. To unravel the possible mechanism(s) involved in this process, microRNA (miR) expression profile, which was assessed using TLDA technique and thereafter confirmed by individual qRT‐PCR, revealed differential expression 10 miRs, five at day 3 and 5 at day 6 post Pi treatment versus control untreated aortas. At day 3, miR‐200c, ‐155, 322 were upregulated and miR‐708 and 331 were downregulated. After 6 days of treatment, miR‐328, ‐546, ‐301a were upregulated whilst miR‐409 and miR‐542 were downregulated. Our results indicate that high Pi levels trigger aortic calcification and modulation of certain miRs. These observations suggest that mechanisms regulating aortic calcification might involve miRs, which warrant further investigations in future studies. This article is protected by copyright. All rights reserved

  August 04, 2017   doi: 10.1002/jcp.26121   open full text
• Statin regulated ERK5 stimulates tight junction formation and reduces permeability in human cardiac endothelial cells.
  Emma L. Wilkinson, James E. Sidaway, Michael J. Cross.
  Journal of Cellular Physiology. August 03, 2017
  The MEKK3/MEK5/ERK5 signaling axis is required for cardiovascular development in vivo. We analyzed the physiological role of ERK5 in cardiac endothelial cells and the consequence of activation of this kinase by the statin class of HMG Co‐A reductase inhibitor drugs. We utilized human cardiac microvascular endothelial cells (HCMECs) and altered ERK5 expression using siRNA mediated gene silencing or overexpression of constitutively active MEK5 and ERK5 to reveal a role for ERK5 in regulating endothelial tight junction formation and cell permeability. Statin treatment of HCMECs stimulated activation of ERK5 and translocation to the plasma membrane resulting in co‐localization with the tight junction protein ZO‐1 and a concomitant reduction in endothelial cell permeability. Statin mediated activation of ERK5 was a consequence of reduced isoprenoid synthesis following HMG Co‐A reductase inhibition. Statin pretreatment could overcome the effect of doxorubicin in reducing endothelial tight junction formation and prevent increased permeability. Our data provide the first evidence for the role of ERK5 in regulating endothelial tight junction formation and endothelial cell permeability. Statin mediated ERK5 activation and the resulting decrease in cardiac endothelial cell permeability may contribute to the cardioprotective effects of statins in reducing doxorubicin‐induced cardiotoxicity. Extracellular signal regulated kinase 5 (ERK5) is required for cardiovascular development in vivo. We have utilized human cardiac endothelial cells to show that ERK5 regulates tight junction formation and permeability in these cells. Statins stimulate ERK5 activity resulting in decreased permeability, which can potentially protect against the adverse effects of cardiotoxic drugs.

  August 03, 2017   doi: 10.1002/jcp.26064   open full text
• The Fam50a positively regulates ameloblast differentiation via interacting with Runx2.
  Yuri Kim, Sung‐Woong Hur, Byung‐Chul Jeong, Sin‐Hye Oh, Yun‐Chan Hwang, Sun‐Hun Kim, Jeong‐Tae Koh.
  Journal of Cellular Physiology. July 17, 2017
  Differentiated ameloblasts secret enamel matrix proteins such as amelogenin, ameloblastin, and enamelin. Expression levels of these proteins are regulated by various factors. To find a new regulatory factor for ameloblast differentiation, we performed 2D‐PAGE analysis using mouse ameloblast lineage cell line (mALCs) cultured with mineralizing medium. Of identified proteins, family with sequence similarity 50 member A (Fam50a) was significantly increased during differentiation of mALCs. Fam50a protein was also highly expressed in secretory ameloblasts of mouse tooth germs. In mALCs cultures, forced expression of Fam50a up‐regulated the expression of enamel matrix protein genes such as amelogenin, ameloblastin, and enamelin. In addition, up‐regulation of Fam50a also increased ALP activity and mineralized nodule formation in a dose‐dependent manner. In contrast, knockdown of Fam50a decreased expression levels of enamel matrix protein genes, ALP activity, and mineralized nodule formation. By fluorescence microscopy, endogenous Fam50a protein was found to be localized to the nucleus of ameloblasts. In addition, Fam50a synergistically increased Ambn transactivation by Runx2. Moreover, Fam50a increased binding affinity of Runx2 to Ambn promoter by physically interacting with Runx2. Taken together, these results suggest Fam50a might be a new positive regulator of ameloblast differentiation. Fam50a protein was selected as a novel regulator of ameloblast differentiation through 2D‐PAGE analysis. Fam50a overexpression stimulated ameloblast differentiation via interacting with Runx2. Our result suggest that Fam50a might be a positive regulator of enamel formation.

  July 17, 2017   doi: 10.1002/jcp.26038   open full text
• TNF‐α has both stimulatory and inhibitory effects on mouse monocyte‐derived osteoclastogenesis.
  Yixuan Cao, Ineke D.C. Jansen, Sara Sprangers, Teun J. de Vries, Vincent Everts.
  Journal of Cellular Physiology. July 17, 2017
  Phenotypically different osteoclasts may be generated from different subsets of precursors. To what extent the formation of these osteoclasts is influenced or mediated by the inflammatory cytokine TNF‐α, is unknown and was investigated in this study. The osteoclast precursors early blasts (CD31hiLy‐6C−), myeloid blasts (CD31+Ly‐6C+), and monocytes (CD31−Ly‐6Chi) were sorted from mouse bone marrow using flow cytometry and cultured with M‐CSF and RANKL, with or without TNF‐α. Surprisingly, TNF‐α prevented the differentiation of TRAcP+ osteoclasts generated from monocytes on plastic; an effect not seen with early blasts and myeloid blasts. This inhibitory effect could not be prevented by other cytokines such as IL‐1β or IL‐6. When monocytes were pre‐cultured with M‐CSF and RANKL followed by exposure to TNF‐α, a stimulatory effect was found. TNF‐α also stimulated monocytes’ osteoclastogenesis when the cells were seeded on bone. Gene expression analysis showed that when TNF‐α was added to monocytes cultured on plastic, RANK, NFATc1, and TRAcP were significantly down‐regulated while TNF‐αR1 and TNF‐αR2 were up‐regulated. FACS analysis showed a decreased uptake of fluorescently labeled RANKL in monocyte cultures in the presence of TNF‐α, indicating an altered ratio of bound‐RANK/unbound‐RANK. Our findings suggest a diverse role of TNF‐α on monocytes’ osteoclastogenesis: it affects the RANK‐signaling pathway therefore inhibits osteoclastogenesis when added at the onset of monocyte culturing. This can be prevented when monocytes were pre‐cultured with M‐CSF and RANKL, which ensures the binding of RANKL to RANK. This could be a mechanism to prevent unfavorable monocyte‐derived osteoclast formation away from the bone. This study describes a multifunctional role of TNF‐α on osteoclastogenesis of a particular subset of osteoclast precursors, the monocytes. TNF‐α proved to prevent osteoclast formation by monocytes when cultured on plastic, while stimulated osteoclastogenesis when the cells were first exposed to M‐CSF and RANKL followed by addition of TNF‐α, or when monocytes were seeded on bone. It is meaningful in modulating osteoclastogenesis in both physiological and inflammatory conditions.

  July 17, 2017   doi: 10.1002/jcp.26024   open full text
• Differential protein modulation by ketoprofen and ibuprofen underlines different cellular response by gastric epithelium.
  L. Brandolini, M. d'Angelo, A. Antonosante, S. Villa, L. Cristiano, V. Castelli, E. Benedetti, M Catanesi, A. Aramini, A. Luini, S Parashuraman, E Mayo, A Giordano, A. Cimini, M. Allegretti.
  Journal of Cellular Physiology. July 15, 2017
  Ketoprofen L‐lysine salt (KLS), is widely used due to its analgesic efficacy and tolerability, and L‐lysine was reported to increase the solubility and the gastric tolerance of ketoprofen. In a recent report, L‐lysine salification has been shown to exert a gastroprotective effect due to its specific ability to counteract the NSAIDs‐induced oxidative stress and up‐regulate gastroprotective proteins. In order to derive further insights into the safety and efficacy profile of KLS, in this study we additionally compared the effect of lysine and arginine, another amino acid counterion commonly used for NSAIDs salification, in control and in ethanol challenged human gastric mucosa model. KLS is widely used for the control of post‐surgical pain and for the management of pain and fever in inflammatory conditions in children and adults. It is generally well tolerated in pediatric patients, and data from three studies in >900 children indicate that oral administration is well tolerated when administered for up to 3 weeks after surgery. Since only few studies have so far investigated the effect of ketoprofen on gastric mucosa maintenance and adaptive mechanisms, in the second part of the study we applied the cMap approach to compare ketoprofen‐induced and ibuprofen‐induced gene expression profiles in order to explore compound‐specific targeted biological pathways. Among the several genes exclusively modulated by ketoprofen, our attention was particularly focused on genes involved in the maintenance of gastric mucosa barrier integrity (cell junctions, morphology and viability). The hypothesis was further validated by Real‐time PCR. This article is protected by copyright. All rights reserved

  July 15, 2017   doi: 10.1002/jcp.26102   open full text
• Visualization of stimulus‐specific heterogeneous activation of individual vascular smooth muscle cells in aortic tissues.
  Satoshi Komatsu, Toshio Kitazawa, Mitsuo Ikebe.
  Journal of Cellular Physiology. July 14, 2017
  Intercellular communication among autonomic nerves, endothelial cells (ECs), and vascular smooth muscle cells (VSMCs) plays a central role in an uninterrupted regulation of blood flow through vascular contractile machinery. Impairment of this communication is linked to development of vascular diseases such as hypertension, cerebral/coronary vasospasms, aortic aneurism, and erectile dysfunction. Although the basic concept of the communication as a whole has been studied, the spatiotemporal correlation of ECs/VSMCs in tissues at the cellular level is unknown. Here, we show a unique VSMC response to ECs during contraction and relaxation of isolated aorta tissues through visualization of spatiotemporal activation patterns of smooth muscle myosin II. ECs in the intimal layer dictate the stimulus‐specific heterogeneous activation pattern of myosin II in VSMCs within distinct medial layers. Myosin light chain (MLC) phosphorylation (active form of myosin II) gradually increases towards outer layers (approximately threefold higher MLC phosphorylation at the outermost layer than that of the innermost layer), presumably by release of an intercellular messenger, nitric oxide (NO). Our study also demonstrates that the MLC phosphorylation at the outermost layer in spontaneously hypertensive rats (SHR) during NO‐induced relaxation is quite high and approximately 10‐fold higher than that of its counterpart, the Wister–Kyoto rats (WKY), suggesting that the distinct pattern of myosin II activation within tissues is important for vascular protection against elevated blood pressure. Impairment of intercellular communication between endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) is linked to development of vascular diseases such as hypertension, cerebral, and coronary vasospasms, aortic aneurism, and erectile dysfunction. Here we show a unique VSMC response to ECs during contraction and relaxation of isolated aorta tissues through visualization of spatiotemporal activation patterns of smooth muscle myosin II.

  July 14, 2017   doi: 10.1002/jcp.25903   open full text
• Apigenin induces ROS‐dependent apoptosis and ER stress in human endometriosis cells.
  Sunwoo Park, Whasun Lim, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. July 14, 2017
  Apigenin is a plant‐derived flavonoid having antiproliferative, anti‐inflammatory, and anti‐angiogenic properties in chronic and metabolic diseases, and cancers. However, the functional role of apigenin remains to be identified in human endometriosis that is a benign inflammatory disease causing infertility, dysmenorrhea, dyspareunia, and chronic abdominal or pelvic pain. In the present study, we determined the effects of apigenin on two well‐established human endometriosis cell lines (VK2/E6E7 and End1/E6E7). Apigenin reduced proliferation and induced cell cycle arrest and apoptosis in the both endometriosis cell lines. In addition, it disrupted mitochondrial membrane potential (MMP) which was accompanied by an increase in concentration of calcium ions in the cytosol and in pro‐apoptotic proteins including Bax and cytochrome c in the VK2/E6E7 and End1/E6E7 cells. Moreover, apigenin treated cells accumulated excessive reactive oxygen species (ROS), and experienced lipid peroxidation and endoplasmic reticulum (ER) stress with activation of the unfolded protein response (UPR) regulatory proteins. Furthermore, the apigenin‐induced apoptosis in endometriosis cells was regulated via the ERK1/2, JNK, and AKT cell signaling pathways. Taken together, apigenin is a potential novel therapeutic agent to overcome current limitations in the treatment to endometriosis. Apigenin is a potential therapeutic substance, which disrupts MMP and induces excessive generation of ROS and ER stress through regulation of the ERK1/2 MAPK, JNK MAPK, and AKT signal transduction pathways, to inhibit proliferation and induce apoptosis in human endometriosis cells.

  July 14, 2017   doi: 10.1002/jcp.26054   open full text
• FHOD1 regulates cytoplasmic actin‐based spindle migration for mouse oocyte asymmetric cell division.
  Meng‐Hao Pan, Fei Wang, Yujie Lu, Feng Tang, Xing Duan, Yu Zhang, Bo Xiong, Shao‐Chen Sun.
  Journal of Cellular Physiology. July 14, 2017
  FHOD1 is a member of Diaphanous related formins (DRFs) which belongs to the Formin family. Previous studies have shown that the DFRs might affect several cellular functions such as morphogenesis, cytokinesis, cell polarity and embryonic differentiation. However, there is no evidence showing the functions of FHOD1 during oocyte meiosis. This study is aimed at exploring the roles of FHOD1 during the mammalian oocyte maturation. Immunofluorescent staining showed that FHOD1 was restricted to the nucleus in germinal vesicle (GV) stage of the oocytes, after the GV breakdown FHOD1 was primarily located at two poles of the spindle at both metaphase I and metaphase II stages. Knockdown of FHOD1 by siRNA injection did not affect polar body extrusion but generated the large polar bodies. In addition, we observed the spindle migration failure in metaphase I oocytes, with a large number of meiotic spindles anchoring in the center of cytoplasm. The expression level of cytoplasmic actin but not cortex actin was significantly reduced, indicating that FHOD1 regulates cytoplasmic actin distribution for the spindle movement. Furthermore, we found that the disruption of ROCK (the Rho‐dependent protein kinase) with inhibitor Y‐27632 caused the decreased FHOD1 protein expression. Therefore, our data indicate that FHOD1 is regulated by ROCK for cytoplasm actin assembly and spindle migration during mouse oocyte meiosis. This article is protected by copyright. All rights reserved

  July 14, 2017   doi: 10.1002/jcp.26099   open full text
• The depletion of MARVELD1 leads to murine placenta accreta via integrin β4‐dependent trophoblast cell invasion.
  Yue Chen, Hui Zhang, Fang Han, Lei Yue, Chunxiao Qiao, Yao Zhang, Peng Dou, Weizhe Liu, Yu Li.
  Journal of Cellular Physiology. July 14, 2017
  The placenta is a remarkable organ, it serves as the interface between the mother and the fetus. Proper invasion of trophoblast cells is required for a successful pregnancy. Previous studies have found that the adhesion molecule integrin β4 plays important roles during trophoblast cell invasion. Here, we found that the overall birth rate of the MARVELD1 knockout mouse is much lower than that of the wild‐type mouse (P<0.001). In E18.5 MARVELD1 knockout mice, we observed an over‐invasion of trophoblast cells, and indeed, the pregnant mice had a partial placenta accreta phenotype. The HTR8/SVneo cell line was used as an in vitro model to elucidate the underlying mechanisms of MARVELD1‐mediated trophoblast invasion. We detected a diminished expression of integrin β4 upon the downregulation of MARVELD1 and enhanced migrate and invasive abilities of trophoblast cells both in vivo and in vitro. The integrin β4 rescue assay also supported the results. In conclusion, this study found that MARVELD1 mediated the invasion of trophoblast cells via regulating the expression of integrin β4 during placenta development. This article is protected by copyright. All rights reserved

  July 14, 2017   doi: 10.1002/jcp.26098   open full text
• GLUT 1 receptor expression and circulating levels of fasting glucose in high grade serous ovarian cancer.
  Laura Pizzuti, Domenico Sergi, Chiara Mandoj, Barbara Antoniani, Francesca Sperati, Andrea Chirico, Luigi Di Lauro, Mario Valle, Alfredo Garofalo, Enrico Vizza, Giacomo Corrado, Federica Tomao, Massimo Rinaldi, Silvia Carpano, Marcello Maugeri‐Saccà, Laura Conti, Giovanna Digiesi, Paolo Marchetti, Ruggero De Maria, Antonio Giordano, Maddalena Barba, Maria A. Carosi, Patrizia Vici.
  Journal of Cellular Physiology. July 14, 2017
  In recent years, the poorly remarkable goals achieved in terms of patients’ important outcomes for ovarian cancer have fueled our interest toward the study of its metabolic roots. Within this research pipeline, we assessed the association between the expression of the glucose transporter GLUT1, as expressed at the tumor tissue level, and circulating pre‐surgical levels of fasting glucose in a case series including data from 40 patients with high FIGO stage serous ovarian cancer. Patients who provided data to the current analysis were randomly selected from a larger cohort. To our purposes, the procedures related to serum and tissue collection, storage and biomarker assessment were highly standardized and centralized at the institutional laboratories. The GLUT1 antibody SPM498 SPRING (REF. E13810) was used at a 1:500 dilution in 2 µm slides. Staining for GLUT1 was observed at the cell membrane level in all the cases assessed, but strong staining was described in 29 (72.5%) of them. The agreement between the two independent reviewers was 100%. Strong GLUT1 staining was inversely associated with circulating levels of fasting glucose, with a particularly striking difference for patients in the lowest fasting glucose tertile (p = 0.044). These results support the biological plausibility of the association of interest. If confirmed in larger studies, our findings may help clarify the potentials of biomarkers related to energy metabolism in terms of prognosis definition, treatment assignment, and outcome interpretation for patients with high FIGO stage serous ovarian cancer. Serous ovarian cystoadenocarcinoma with intense positive staining for glucose transporter protein 1 (GLUT1). A. GLUT1 x20; B. GLUT1x10.

  July 14, 2017   doi: 10.1002/jcp.26023   open full text
• The Xenoestrogens Biphenol‐A and Nonylphenol differentially regulate metalloprotease‐mediated shedding of EGFR ligands.
  Paulina Urriola‐Muñoz, Xue Li, Thorsten Maretzky, David R. McIlwain, Tak W. Mak, Juan G Reyes, Carl P. Blobel, Ricardo D. Moreno.
  Journal of Cellular Physiology. July 13, 2017
  The xenoestrogens Bisphenol‐A (BPA) and Nonylphenol (NP) are endocrine disruptors used in the plastic polymer industry to manufacture different products for human use. Previous studies have suggested a role of these compounds in the shedding of signaling molecules, such as tumor necrosis factor α (TNF‐ α). The aim of this work was to evaluate the effect of BPA and NP on the sheddase ADAM17 and its newly discovered regulators iRhom1 and iRhom2 in the release of EGFR‐ligands. We report that BPA and NP can stimulate the release of the ADAM17‐substrates HB‐EGF and TGF‐ α. In cells lacking ADAM17 (Adam17‐/‐ mEFs) BPA‐stimulated release of HB‐EGF, but not TGF‐ α, was strongly reduced, whereas NP‐stimulated shedding of HB‐EGF and TGF‐ α was completely abolished. Inactivation of both ADAM17 and the related ADAM10 (Adam10/17‐/‐ mEFs) completely prevented the release of these substrates. In the absence of iRhom1, BPA‐ or NP‐stimulated release of HB‐EGF or TGF‐ α was comparable to wild type control mEFs, conversely the BPA‐induced release of HB‐EGF was abolished in iRhom2‐/‐ mEFs. The defect in shedding of HB‐EGF in iRhom2‐/‐ mEF cells could be rescued by overexpressing iRhom2. Interestingly, the NP‐stimulated release of HB‐EGF was not affected by the absence of iRhom2, suggesting that NP could potentially activate both ADAM10 and ADAM17. We tested this hypothesis using betacellulin (BTC), an EGFR‐ligand that is a substrate for ADAM10. We found that NP, but not BPA stimulated the release of BTC in Adam17‐/‐, iRhom2‐/‐ or iRhom1/2‐/‐, but not in Adam10/17‐/‐ cells. Taken together, our results suggest that BPA and NP stimulate the release of EGFR‐ligands by differentially activating ADAM17 or ADAM10. The identification of specific effects of these endocrine disruptors on ADAM10 and ADAM17 will help to provide a better understanding of their roles in cell signaling and proinflammatory processes, and provide new potential targets for treatment of reproductive or inflammatory diseases such as asthma or breast cancer that are promoted by xenoestrogens. This article is protected by copyright. All rights reserved

  July 13, 2017   doi: 10.1002/jcp.26097   open full text
• Fasting inhibits hepatic stellate cells activation and potentiates anti‐cancer activity of Sorafenib in hepatocellular cancer cells.
  Oriana Lo Re, Concetta Panebianco, Stefania Porto, Carlo Cervi, Francesca Rappa, Stefano Di Biase, Michele Caraglia, Valerio Pazienza, Manlio Vinciguerra.
  Journal of Cellular Physiology. July 11, 2017
  Hepatocellular carcinoma (HCC) has a poor outcome. Most HCCs develop in the context of liver fibrosis and cirrhosis caused by chronic inflammation. Short‐term fasting approaches enhance the activity of chemotherapy in preclinical cancer models, other than HCC. Multi‐tyrosine kinase inhibitor Sorafenib is the mainstay of treatment in HCC. However, its benefit is frequently short‐lived. Whether fasting can alleviate liver fibrosis and whether combining fasting with Sorafenib is beneficial remains unknown. A 24 hr fasting (2% serum, 0.1% glucose)‐induced changes on human hepatic stellate cells (HSC) LX‐2 proliferation/viability/cell cycle were assessed by MTT and flow cytometry. Expression of lypolysaccharide (LPS)‐induced activation markers (vimentin, αSMA) was evaluated by qPCR and immunoblotting. Liver fibrosis and inflammation were evaluated in a mouse model of steatohepatitis exposed to cycles of fasting, by histological and biochemical analyses. A 24 hr fasting‐induced changes were also analyzed on the proliferation/viability/glucose uptake of human HCC cells exposed to Sorafenib. An expression panel of genes involved in survival, inflammation, and metabolism was examined by qPCR in HCC cells exposed to fasting and/or Sorafenib. Fasting decreased the proliferation and the activation of HSC. Repeated cycles of short term starvation were safe in mice but did not improve fibrosis. Fasting synergized with Sorafenib in hampering HCC cell growth and glucose uptake. Finally, fasting normalized the expression levels of genes which are commonly altered by Sorafenib in HCC cells. Fasting or fasting‐mimicking diet diets should be evaluated in preclinical studies as a mean to potentiate the activity of Sorafenib in clinical use. Short‐term fasting (STS) cycles inhibit hepatic stellate cells activation and hepatocellular carcinoma cells proliferation in vitro. STS are harmless in an in vivo model of liver fibrosis. STS cycles might be promising approaches to treat liver diseases.

  July 11, 2017   doi: 10.1002/jcp.25987   open full text
• Three‐dimensional cell culture models for anticancer drug screening: Worth the effort?
  Eddy‐Tim Verjans, Jordi Doijen, Walter Luyten, Bart Landuyt, Liliane Schoofs.
  Journal of Cellular Physiology. July 11, 2017
  High attrition of new oncology drug candidates in clinical trials is partially caused by the poor predictive capacity of artificial monolayer cell culture assays early in drug discovery. Monolayer assays do not take the natural three‐dimensional (3D) microenvironment of cells into account. As a result, false positive compounds often enter clinical trials, leading to high dropout rates and a waste of time and money. Over the past 2 decades, tissue engineers and cell biologists have developed a broad range of 3D in vitro culturing tools that better represent in vivo cell biology. These tools preserve the 3D architecture of cells and can be used to predict toxicity of and resistance against antitumor agents. Recent progress in tissue engineering further improves 3D models by taking into account the tumor microenvironment, which is important for metastatic progression and vascularization. However, the widespread implementation of 3D cell cultures into cell‐based research programs has been limited by various factors, including their cost and reproducibility. In addition, different 3D cell culture techniques often produce spheroids of different size and shape, which can strongly influence drug efficacy and toxicity. Hence, it is imperative to morphometrically characterize multicellular spheroids to avoid generalizations among different spheroid types. Standardized 3D culturing procedures could further reduce data variability and enhance biological relevance. Here, we critically evaluate the benefits and challenges inherent to growing cells in 3D, along with an overview of the techniques used to form spheroids. This is done with a specific focus on antitumor drug screening. Over the past few decades, a multitude of 3D cell culturing techniques have been developed of which some have proven to be valuable for cancer research. Their implementation, however, has been slowed down due to extensive variation between experimental outcomes when using different culturing systems. We provide an overview of the current 3D culturing techniques while also addressing the underlying cause of variation and the requirements before widespread implementation of 3D cell cultures for drug discovery can occur.

  July 11, 2017   doi: 10.1002/jcp.26052   open full text
• Lipoprotein(a): A missing culprit in the management of athero‐thrombosis?
  Gianna Ferretti, Tiziana Bacchetti, Thomas P. Johnston, Maciej Banach, Matteo Pirro, Amirhossein Sahebkar.
  Journal of Cellular Physiology. July 11, 2017
  Lipoprotein(a) Lp(a) is a cholesterol‐rich, LDL‐like particle that is independently associated with an increased risk for ischemic heart disease, atherosclerosis, thrombosis, and stroke. Genetic variation in the Lp(a) locus and some other genes related to Lp(a) synthesis and metabolism play a critical role in regulating plasma Lp(a) levels. The pathophysiological potential of Lp(a) is related to proatherogenic and prothrombotic effects on the vasculature. Different molecular mechanisms underlying the atherothrombotic potential of Lp(a), free apolipoprotein(a), and oxidized‐Lp(a) have been proposed. However, plasma Lp(a) assay is complicated by problems associated with quantification and standardization owing to the polymorphic nature of this lipoprotein. This review has focused on the physicochemical properties of Lp(a), the genetic aspects of Lp(a), the need for accurate determination of Lp(a), the synthesis, and recent findings on metabolism of Lp(a). Lastly, the patho‐physiological mechanisms by which Lp(a) may increase athero‐thrombosis and an overview on the therapeutic modalities to interfere with Lp(a) are summarized. This review has focused on the physicochemical properties of Lp(a), the genetic aspects of Lp(a), the need for accurate determination of Lp(a), the synthesis, and recent findings on metabolism of Lp(a). The patho‐physiological mechanisms by which Lp(a) may increase athero‐thrombosis and an overview on the therapeutic modalities to interfere with Lp(a) are summarized.

  July 11, 2017   doi: 10.1002/jcp.26050   open full text
• Poly r(C) binding protein 1‐mediated regulation of microRNA expression underlies post‐sevoflurane amelioration of acute lung injury in rats.
  Xiaohong Zhao, Xiaoxia Wang, Fei Wang, Chengjie Gao, Jian Wang.
  Journal of Cellular Physiology. July 11, 2017
  Acute lung injury (ALI) presents a pervasive health burden due to the high morbidity and mortality associated with it. Volatile anesthetics like sevoflurane has been previously shown to have organ‐protective effect, both in the context of normal physiological function in liver, and during LPS‐induced ALI. Sevoflurane was shown to exert lung protective effect during LPS‐induced ALI by modulating expression level of microRNAs (miRNAs), specifically miR‐155. The objective of the current study was to define the underlying mechanism by which sevoflurane alters miRNA expression levels. Lung injury caused by LPS and its amelioration post sevoflurane administration was first confirmed. Expression levels of different miRNA and messenger RNAs (mRNAs) encoding inflammatory cytokines were measured in a rat model of lipopolysaccharide (LPS)‐induced ALI, which were subsequently treated with either sevoflurane or vehicle control. Host of miRNAs and messenger RNAs encoding pro‐inflammatory cytokines are overexpressed during LPS‐induced ALI, which are reversed following sevoflurane administration. Mass spectrometry analysis revealed that the RNA‐binding protein, poly r(C) binding protein 1 (PCBP1) expression is induced in ALI and is repressed following sevoflurane treatment. RNA immunoprecipitation experiments revealed that PCBP1 expression dictates the altered miRNA expression and sevoflurane altered miRNA expression by suppressing PCBP1 expression. Our study thus elucidates a unique mechanism of lung protective effect of sevoflurane mediated by suppression of expression of a RNA‐binding protein that potentiates expression of pro‐inflammatory miRNAs. Our study thus elucidates a unique mechanism of lung protective effect of sevoflurane mediated by suppression of expression of a RNA‐binding protein that potentiates expression of pro‐inflammatory miRNAs.

  July 11, 2017   doi: 10.1002/jcp.26053   open full text
• Osteogenic commitment and differentiation of human mesenchymal stem cells by low‐intensity pulsed ultrasound stimulation.
  Viviana Costa, Valeria Carina, Simona Fontana, Angela De Luca, Francesca Monteleone, Stefania Pagani, Maria Sartori, Stefania Setti, Cesare Faldini, Riccardo Alessandro, Milena Fini, Gianluca Giavaresi.
  Journal of Cellular Physiology. July 11, 2017
  Low‐intensity pulsed ultrasound (LIPUS) as an adjuvant therapy in in vitro and in vivo bone engineering has proven to be extremely useful. The present study aimed at investigating the effect of 30 mW/cm2 LIPUS stimulation on commercially available human mesenchymal stem cells (hMSCs) cultured in basal or osteogenic medium at different experimental time points (7, 14, 21 days). The hypothesis was that LIPUS would improve the osteogenic differentiation of hMSC and guarantying the maintenance of osteogenic committed fraction, as demonstrated by cell vitality and proteomic analysis. LIPUS stimulation (a) regulated the balance between osteoblast commitment and differentiation by specific networks (activations of RhoA/ROCK signaling and upregulation of Ribosome constituent/Protein metabolic process, Glycolysis/Gluconeogenesis, RNA metabolic process/Splicing and Tubulins); (b) allowed the maintenance of a few percentage of osteoblast precursors (21 days CD73+/CD90+: 6%; OCT‐3/4+/NANOG+/SOX2+: 10%); (c) induced the activation of osteogenic specific pathways shown by gene expression (early: ALPL, COL1A1, late: RUNX2, BGLAP, MAPK1/6) and related protein release (COL1a1, OPN, OC), in particular in the presence of osteogenic soluble factors able to mimic bone microenvironment. To summarize, LIPUS might be able to improve the osteogenic commitment of hMSCs in vitro, and, at the same time, enhance their osteogenic differentiation. LIPUS (30 mW/cm2) might be able to maintain hMSC stemness during osteogenic differentiation, guarantying the undifferentiated state of a fraction of them. It regulates the balance between stemness maintenance and osteoblast differentiation by specific proteomic networks. LIPUS mallows the maintenance of a few percentage of multipotent hMSCs, activating osteogenic specific pathways.

  July 11, 2017   doi: 10.1002/jcp.26058   open full text
• Bryostatin and its synthetic analog, picolog rescue dermal fibroblasts from prolonged stress and contribute to survival and rejuvenation of human skin equivalents.
  Tapan K. Khan, Paul A. Wender, Daniel L. Alkon.
  Journal of Cellular Physiology. July 11, 2017
  Skin health is associated with the day‐to‐day activity of fibroblasts. The primary function of fibroblasts is to synthesize structural proteins, such as collagen, extracellular matrix proteins, and other proteins that support the structural integrity of the skin and are associated with younger, firmer, and more elastic skin that is better able to resist and recover from injury. At sub‐nanomolar concentrations (0.03–0.3 nM), bryostatin‐1 and its synthetic analog, picolog (0.1–10 nM) sustained the survival and activation of human dermal fibroblasts cultured under the stressful condition of prolonged serum deprivation. Bryostatin‐1 treatment stabilized human skin equivalents (HSEs), a bioengineered combination of primary human skin cells (keratinocytes and dermal fibroblasts) on an extracellular matrix composed of mainly collagen. Fibroblasts activated by bryostatin‐1 protected the structural integrity of HSEs. Bryostatin‐1 and picolog prolonged activation of Erk in fibroblasts to promote cell survival. Chronic stress promotes the progression of apoptosis. Dermal fibroblasts constitutively express all components of Fas associated apoptosis, including caspase‐8, an initiator enzyme of apoptosis. Prolong bryostatin‐1 treatment reduced apoptosis by decreasing caspase‐8 and protected dermal fibroblasts. Our data suggest that bryostatin‐1 and picolog could be useful in anti‐aging skincare, and could have applications in tissue engineering and regenerative medicine. At sub‐nanomolar concentrations (0.03–0.3 nM), bryostatin‐1 and its synthetic analog, picolog (0.1–10 nM) sustained the survival and activation of human dermal fibroblasts cultured under the stressful condition of prolonged serum deprivation. Bryostatin‐1 treatment stabilized human skin equivalents (HSEs), a bioengineered combination of primary human skin cells (keratinocytes and dermal fibroblasts) on an extracellular matrix composed of mainly collagen. Fibroblasts activated by bryostatin‐1 protected the structural integrity of HSEs.

  July 11, 2017   doi: 10.1002/jcp.26043   open full text
• Integrated transcriptomic and metabolomic analysis reveals adaptive changes of hibernating retinas.
  Yizhao Luan, Jingxing Ou, Vincent P. Kunze, Fengyu Qiao, Yan Wang, Lai Wei, Wei Li, Zhi Xie.
  Journal of Cellular Physiology. July 11, 2017
  Hibernation is a seasonally adaptive strategy that allows hibernators to live through extremely cold conditions. Despite the profound reduction of blood flow to the retinas, hibernation causes no lasting retinal injury. Instead, hibernators show an increased tolerance to ischemic insults during the hibernation period. To understand the molecular changes of the retinas in response to hibernation, we applied an integrative transcriptome and metabolome analysis to explore changes in gene expression and metabolites of 13‐lined ground squirrel retinas during hibernation. Metabolomic analysis showed a global decrease of ATP synthesis in hibernating retinas. Decreased glucose and galactose, increased beta‐oxidation of carnitine and decreased storage of some amino acids in hibernating retinas indicated a shift of fuel use from carbohydrates to lipids and alternative usage of amino acids. Transcriptomic analysis revealed that the down‐regulated genes were enriched in DNA‐templated transcription and immune‐related functions, while the up‐regulated genes were enriched in mitochondrial inner membrane and DNA packaging‐related functions. We further showed that a subset of genes underwent active alternative splicing events in response to hibernation. Finally, integrative analysis of the transcriptome and metabolome confirmed the shift of fuel use in the hibernating retina by the regulation of catabolism of amino acids and lipids. Through transcriptomic and metabolomic data, our analysis revealed the altered state of mitochondrial oxidative phosphorylation and the shift of energy source in the hibernating retina, advancing our understanding of the molecular mechanisms employed by hibernators. The data will also serve as a useful resource for the ocular and hibernation research communities. To understand the molecular changes of the retinas in response to hibernation, we applied an integrative transcriptome and metabolome analysis to explore changes in gene expression and metabolites of 13‐lined ground squirrel retinas during hibernation. Metabolomic analysis showed a global decrease of ATP synthesis and a shift of fuel use from carbohydrates to lipids and alternative usage of amino acids in hibernating retinas. Transcriptomic analysis revealed that the down‐regulated genes were enriched in DNA‐templated transcription and immune‐related functions, while the up‐regulated genes were enriched in mitochondrial inner membrane and DNA packaging‐related functions.

  July 11, 2017   doi: 10.1002/jcp.26030   open full text
• Wnt5a Suppresses Osteoblastic Differentiation of Human Periodontal Ligament Stem Cell‐like Cells Via Ror2/JNK signaling.
  Daigaku Hasegawa, Naohisa Wada, Shinichiro Yoshida, Hiromi Mitarai, Mai Arima, Atsushi Tomokiyo, Sayuri Hamano, Hideki Sugii, Hidefumi Maeda.
  Journal of Cellular Physiology. July 06, 2017
  Wnt5a, a non‐canonical Wnt protein, is known to play important roles in several cell functions. However, little is known about the effects of Wnt5a on osteoblastic differentiation of periodontal ligament (PDL) cells. Here, we examined the effects of Wnt5a on osteoblastic differentiation and associated intracellular signaling in human PDL stem/progenitor cells (HPDLSCs). We found that Wnt5a suppressed expression of bone‐related genes (ALP, BSP, and Osterix) and alizarin red‐positive mineralized nodule formation in HPDLSCs under osteogenic conditions. Immunohistochemical analysis revealed that a Wnt5a‐related receptor, receptor tyrosine kinase‐like orphan receptor 2 (Ror2), was expressed in rat PDL tissue. Interestingly, knockdown of Ror2 by siRNA inhibited the Wnt5a‐induced downregulation of bone‐related gene expression in HPDLSCs. Moreover, western blotting analysis showed that phosphorylation of the intracellular signaling molecule, c‐Jun N‐terminal kinase (JNK) was upregulated in HPDLSCs cultured in osteoblast induction medium with Wnt5a, but knockdown of Ror2 by siRNA downregulated the phosphorylation of JNK. We also examined the effects of JNK inhibition on Wnt5a‐induced suppression of osteoblastic differentiation of HPDLSCs. The JNK inhibitor, SP600125 inhibited the Wnt5a‐induced downregulation of bone‐related gene expression in HPDLSCs. Additionally, SP600125 inhibited the Wnt5a‐induced suppression of the alizarin red‐positive reaction in HPDLSCs. These results suggest that Wnt5a suppressed osteoblastic differentiation of HPDLSCs through Ror2/JNK signaling. Non‐canonical Wnt signaling, including Wnt5a/Ror2/JNK signaling, may function as a negative regulator of mineralization, preventing the development of non‐physiological mineralization in PDL tissue. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26086   open full text
• Targeting P‐glycoprotein and SORCIN: Dihydromyricetin strengthens anti‐proliferative efficiency of adriamycin via MAPK/ERK and Ca2+‐mediated apoptosis pathways in MCF‐7/ADR and K562/ADR.
  Yaoting Sun, Changyuan Wang, Qiang Meng, Zhihao Liu, Xiaokui Huo, Pengyuan Sun, Huijun Sun, Xiaodong Ma, Jinyong Peng, Kexin Liu.
  Journal of Cellular Physiology. July 06, 2017
  Recently, a new target Ca2+‐binding protein SORCIN was reported to participate in multidrug resistance (MDR) in cancer. Here we aim to investigate whether dihydromyricetin (DMY), a dihydroflavonol compound with anti‐inflamatory, anti‐oxidant, anti‐bacterial and anti‐tumor actions, reverses MDR in MCF‐7/ADR and K562/ADR and to elucidate its potential molecular mechanism. DMY enhanced cytotoxicity of adriamycin (ADR) by downregulating MDR1 mRNA and P‐gp expression through MAPK/ERK pathway and also inhibiting the function of P‐gp significantly. Meanwhile, DMY decreased mRNA and protein expression of SORCIN, which resulted in elevating intracellular free Ca2+. Finally, we investigated co‐administration ADR with DMY remarkably increased ADR‐induced apoptosis. Further study showed DMY elevated ROS levels and caspase‐12 protein expression, which signal apoptosis in endoplasmic reticulum. At the same time, proteins related to mitochondrial apoptosis were also changed such as Bcl‐2, Bax, caspase‐3, caspase‐9 and PARP. Finally, nude mice model also demonstrated that DMY strengthened anti‐tumor activity of ADR in vivo. In conclusion, DMY reverses MDR by downregulating P‐gp, SORCIN expression and increasing free Ca2+, as well as, inducing apoptosis in MCF‐7/ADR and K562/ADR. These fundamental findings provide evidence for further clinical research in application of DMY as an assistant agent in the treatment of cancer. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26087   open full text
• Concentration‐Dependent Metabolic Effects of Metformin in Healthy and Fanconi Anemia Lymphoblast Cells.
  Silvia Ravera, Vanessa Cossu, Barbara Tappino, Elena Nicchia, Carlo Dufour, Simona Cavani, Andrea Sciutto, Claudia Bolognesi, Marta Columbaro, Paolo Degan, Enrico Cappelli.
  Journal of Cellular Physiology. July 06, 2017
  Metformin (MET) is the drug of choice for patients with type 2 diabetes and has been proposed for use in cancer therapy and for treating other metabolic diseases. More than 14,000 studies have been published addressing the cellular mechanisms affected by MET. However, several in vitro studies have used concentrations of the drug 10–100‐fold higher than the plasmatic concentration measured in patients. Here we evaluated the biochemical, metabolic and morphologic effects of various concentrations of MET. Moreover, we tested the effect of MET on Fanconi Anemia (FA) cells, a DNA repair genetic disease with defects in energetic and glucose metabolism, as well as on human promyelocytic leukemia (HL60) cell lines. We found that the response of wild‐type cells to MET is concentration dependent. Low concentrations (15 and 150 µM) increase both oxidative phosphorylation and the oxidative stress response, acting on the AMPK/Sirt1 pathway, while the high concentration (1.5 mM) inhibits the respiratory chain, alters cell morphology, becoming toxic to the cells. In FA cells, MET was unable to correct the energetic/respiratory defect and did not improve the response to oxidative stress and DNA damage. By contrast, HL60 cells appear sensitive also at 150 µM. Our findings underline the importance of the MET concentration in evaluating the effect of this drug on cell metabolism and demonstrate that data obtained from in vitro experiments, that have used high concentrations of MET, cannot be readily translated into improving our understanding of the cellular effects of metformin when used in the clinical setting. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26085   open full text
• 17β‐estradiol rescues damages following traumatic brain injury from molecule to behavior in mice.
  Huaihai Lu, Kun Ma, Liwei Jin, He Zhu, Ruiqi Cao.
  Journal of Cellular Physiology. July 06, 2017
  Traumatic brain injury (TBI) is a public health concern, and causes cognitive dysfunction, emotional disorders, and neurodegeration, as well. The currently available treatments are all symptom‐oriented with unsatifying efficacy. It is highly demanded to understand its underlying mechanisms. Controlled cortical impact (CCI) was used to induce TBI in aged female mice subjected to ovariectomy. Brain damages were assessed with neurological severity score, brain infarction and edema. Morris water maze and elevated plus maze were applied to evaluate the levels of anxiety. Apoptosis in the hippocampus was assayed with Fluoro‐Jade B staining and TUNEL staining. Western blot was employed to measure the expression of NMDA receptor subunits and phosphorylation of ERK1/2, and biochemical assays were used to estimate oxidative stress. 17beta‐Estradiol (E2) was intraperitoneally administered at 10‐80 µg/kg once per day for 7 consecutive days before or after CCI. Chronic administration of E2 both before and immediately after CCI conferred neuroprotection, reducing neurological severity score, brain infarction and edema in TBI mice. Additionally, E2 improved many aspects of deleterious effects of TBI on the hippocampus, including neuronal apoptosis, dysfunction in spatial memory, reduction in NR2B, enhancement of oxidative stress, and activation of ERK1/2 pathway. The present study provides clue for the notion that E2 has therapeutic potential for both prevention and intervention of TBI‐induced brain damages. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26083   open full text
• Occludin as a functional marker of vascular endothelial cells on tube‐forming activity.
  Toshie Kanayasu‐Toyoda, Akiko Ishii‐Watabe, Yutaka Kikuchi, Hiroko Kitagawa, Hiroko Suzuki, Hiroomi Tamura, Minoru Tada, Takuo Suzuki, Hiroyuki Mizuguchi, Teruhide Yamaguchi.
  Journal of Cellular Physiology. July 06, 2017
  Cell therapy using endothelial progenitor cells (EPCs) is a promising strategy for the treatment of ischemic diseases. Two types of EPCs have been identified: early EPCs and late EPCs. Late EPCs are able to form tube structure by themselves, and have a high proliferative ability. The functional marker(s) of late EPCs, which relate to their therapeutic potential, have not been fully elucidated. Here we compared the gene expression profiles of several human cord blood derived late EPC lines which exhibit different tube formation activity, and we observed that the expression of occludin (OCLN) in these lines correlated with the tube formation ability, suggesting that OCLN is a candidate functional marker of late EPCs. When OCLN was knocked down by transfecting siRNA, the tube formation on Matrigel, the S phase + G2/M phase in the cell cycle, and the spheroid‐based sprouting of late EPCs were markedly reduced, suggesting the critical role of OCLN in tube formation, sprouting and proliferation. These results indicated that OCLN plays a novel role in neovascularization and angiogenesis. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26082   open full text
• Luteoloside Prevents Lipopolysaccharide‐Induced Osteolysis and Suppresses RANKL‐Induced Osteoclastogenesis Through Attenuating RANKL Signaling Cascades.
  Fangming Song, Chengming Wei, Lin Zhou, An Qin, Mingli Yang, Jennifer Tickner, Yuanjiao Huang, Jinmin Zhao, Jiake Xu.
  Journal of Cellular Physiology. July 06, 2017
  Bone destruction or osteolysis marked by excessive osteoclastic bone resorption is a very common medical condition. Identification of agents that can effectively suppress excessive osteoclast formation and function is crucial for prevention and treatment of osteolytic conditions such as periprosthetic joint infection and periprosthetic loosening. Luteoloside, a flavonoid, is a natural bioactive compound with anti‐inflammation and anti‐tumor properties. However, the effect of Luteoloside on inflammation‐induced osteolysis is unknown. Here, we found that Luteoloside exhibited a strong inhibitory effect on lipopolysaccharide (LPS)‐induced osteolysis in vivo. In addition, Luteoloside suppressed RANKL‐induced osteoclast differentiation and abrogated bone resorption in a dose‐dependent manner. Further, we found that the anti‐osteoclastic and anti‐resorptive actions of Luteoloside are mediated via blocking NFATc1 activity and the attenuation of RANKL‐mediated Ca2+ signaling as well as NF‐κB and MAPK pathways. Taken together, this study shows that Luteoloside may be a potential therapeutic agent for osteolytic bone diseases associated with abnormal osteoclast formation and function in inflammatory conditions. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26084   open full text
• Indomethacin Elicits Proteasomal Dysfunctions Develops Apoptosis Through Mitochondrial Abnormalities.
  Ayeman Amanullah, Ribhav Mishra, Arun Upadhyay, P. Purushotham Reddy, Ranabir Das, Amit Mishra.
  Journal of Cellular Physiology. July 06, 2017
  Non‐steroidal anti‐inflammatory drugs (NSAIDs) are a class of drugs that are mainly used to treat pain, inflammation and fever via cyclooxygenase‐2 (COX‐2) inhibition. There are abundant findings that uncover the hidden critical chemotherapeutics potential of NSAIDs in cancer treatment. However, still the precise mechanism by which NSAIDs could be used as an effective anti‐tumor agent in the prevention of carcinogenesis is not well understood. Here, we show that indomethacin, a well‐known NSAID, induces proteasomal dysfunction that results in accumulation of unwanted proteins, mitochondrial abnormalities and successively stimulate apoptosis in cells. We observed the interaction of indomethacin with proteasome and noticed the massive accumulation of intracellular ubiquitin‐positive proteins, which might be due to the suppression of proteasome activities. Furthermore, we also found that exposure of indomethacin causes the accumulation of critical proteasomal substrates that consequently generate severe mitochondrial abnormalities and prompt up key apoptotic events in cells. Our results demonstrate how indomethacin affects normal proteasomal functions and induces mitochondrial apoptosis in cells. These findings also improve our current understanding of how NSAIDs can exhibit crucial anti‐proliferative effects in cells. In near future, our findings may suggest a new possible strategy for the development of specific proteasome inhibitors in conjunction with other chemo‐preventive anticancer agents. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26081   open full text
• 3,4‐dihydroxybenzalacetone and caffeic acid phenethyl ester induce preconditioning ER stress and autophagy in SH‐SY5Y cells.
  Ryoichi Tomiyama, Ken Takakura, Shouhei Takatou, Thuong Manh Le, Takumi Nishiuchi, Yutaka Nakamura, Tetsuya Konishi, Seiichi Matsugo, Osamu Hori.
  Journal of Cellular Physiology. July 06, 2017
  3,4‐dihydroxybenzalacetone (DBL) and Caffeic acid phenethyl ester (CAPE) are both catechol‐containing phenylpropanoid derivatives with diverse bioactivities. In the present study, we analyzed the ability of these compounds to activate the unfolded protein response (UPR) and the oxidative stress response. When human SH‐SY5Y neuroblastoma cells were treated with DBL or CAPE, the expression of endoplasmic reticulum (ER) stress‐related genes such as HSPA5, HYOU1, DDIT3, and SEC61b increased to a larger extent in response to CAPE treatment, while that of antioxidant genes such as HMOX1, GCLM, and NQO1 increased to a larger extent in response to DBL treatment. DNA microarray analysis confirmed the strong link of these compounds to ER stress. Regarding the mechanism, activation of the UPR by these compounds was associated with enhanced levels of oxidized proteins in the ER, and N‐acetyl cysteine (NAC), which provides anti‐oxidative effects, suppressed the induction of the UPR‐target genes. Furthermore, both compounds enhanced the expression of LC3‐II, a marker of autophagy, and 4‐Phenylbutyric acid (4‐PBA), a chemical chaperone that reduces ER stress, suppressed it. Finally, pretreatment of cells with DBL, CAPE or low doses of ER stressors protected cells against a neurotoxin 6‐hydroxydopamine (6‐OHDA) in an autophagy‐dependent manner. These results suggest that DBL and CAPE induce oxidized protein‐mediated ER stress and autophagy that may have a preconditioning effect in SH‐SY5Y cells. This article is protected by copyright. All rights reserved

  July 06, 2017   doi: 10.1002/jcp.26080   open full text
• What role does the stress response have in congestive heart failure?
  Ahmed Badreddin, Youssef Fady, Hamdy Attia, Mohamed Hafez, Ahmed Khairallah, Dina Johar, Larry Bernstein.
  Journal of Cellular Physiology. July 04, 2017
  This review is concerned with cardiac malfunction as a result of an imbalance in protein proteostasis, the homeostatic balance between protein removal and regeneration in a long remodeling process involving the endoplasmic reticulum (ER) and the unfolded protein response (UPR). The importance of this is of special significance with regard to cardiac function as a high energy requiring muscular organ that has a high oxygen requirement and is highly dependent on mitochondria. The importance of mitochondria is not only concerned with high energy dependence on mitochondrial electron transport, but it also has a role in the signaling between the mitochondria and the ER under stress. Proteins made in the ER are folded as a result of sulfhydryl groups (−SH) and attractive and repulsive reactions in the tertiary structure. We discuss how this matters with respect to an imbalance between muscle breakdown and repair in a stressful environment, especially as a result of oxidative and nitrosative byproducts of mitochondrial activity. The normal repair is a remodeling, but under this circumstance, the cell undergoes or even lysosomal “self eating” autophagy, or even necrosis instead of apoptosis. We shall discuss the relationship of the UPR pathway to chronic congestive heart failure (CHF). The left ventricular wall thickening is correlated with phosphorylated eIF2/eIF2 ratio and level of dysregulation of calcium‐handling proteins, including, P‐ryanodine receptor, Ca2+ storing protein calsequestrin, Na‐Ca2+ exchanger, sarcoendoplasmic reticulum Ca2+‐ATPase (SERCA2α), ER chaperone protein calreticulin. The UPR occurs as a protein unfolding with the breaking of sulfhydryl groups in the tertiary structure. If the stress on proteostasis is for an extended period, the ability to repair may be compromised. Consequently, the opened protein skeletons may aggregate and when the stress is sufficient, the transport of protein is disabled and protein occludes the ER canal. GRP78 (also known as BiP) is a heat‐shock protein chaperone resident in the ER that binds and blocks three sensor proteins present in the ER membrane in the basal stress conditions. In unstressed conditions, binds to PERK, ATF6, and inositol‐requiring protein‐1α (IRE1α). GRP78 shifts from blocking the sensor proteins to binding the unfolded proteins, triggering UPR when unfolded protein accumulates in the ER lumen. When there is a mild stress, unfolded protein accumulates, and universal protein synthesis is inhibited and it resumes upon recovery. Activated IRE1α then splices X‐box binding protein‐1 (XBP‐1) mRNA, which leads to synthesis of the active transcription factor XBP‐1 and upregulation of UPR genes. Active ATF6 translocates from the ER to Golgi where it activates genes to increase ER protein folding. Once activated three sensor proteins PERK, IRE1α, and ATF6 lead to inhibition of translation and increased transcription of chaperones, stress response genes, and redox‐related genes. PERK phosphorylates the translation initiation factor eIF2α, thereby inhibiting global translation and reducing the load of newly synthesized. eIF2α phosphorylation allows ATF4 translation, a transcriptional activator that induces a cascade that ultimately produces proapoptotic factors. A more severe or persistent stress would lead to apoptosis.

  July 04, 2017   doi: 10.1002/jcp.26003   open full text
• PCSK9 and infection: A potentially useful or dangerous association?
  Farzad Khademi, Amir Abbas Momtazi‐borojeni, Željko Reiner, Maciej Banach, Khalid Al Al‐Rasadi, Amirhossein Sahebkar.
  Journal of Cellular Physiology. July 04, 2017
  Elevated plasma low‐density lipoprotein‐cholesterol (LDL‐C) concentration is the most important risk factor for atherosclerotic cardiovascular diseases (CVDs). Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a ubiquitously expressed serine proteinase which plays a key role in cholesterol metabolism, but has been found to be implicated in some other lipid‐independent physiological processes. In this review, the role of PCSK9 was evaluated not only concerning lipid metabolism but also hepatitis C virus (HCV) infection, bacterial infections/sepsis, and septic shock. Collected data from clinical trials revealed that treatment with PCSK9 inhibitors has beneficial effects in lowering LDL‐C via inhibition of LDL‐receptors (LDL‐R), an antiviral effect on HCV infection via down‐regulating the surface expression of LDL‐R and CD81 on hepatic cells, and a positive association with increased inflammatory responses, as well as with septic shock by down‐regulation of hepatocyte LDL‐R. On the other hand, PCSK9 inhibition by therapeutic fully humanized antibodies has positive effects in reducing elevated LDL‐C. However, their safety and tolerability is an important issue which has to be taken into consideration. In this review, the role of PCSK9 was evaluated not only concerning lipid metabolism but also hepatitis C virus (HCV) infection, bacterial infections/sepsis, and septic shock.

  July 04, 2017   doi: 10.1002/jcp.26040   open full text
• The significant role of the Golgi apparatus in cardiovascular diseases.
  Liqun Lu, Qun Zhou, Zhe Chen, Linxi Chen.
  Journal of Cellular Physiology. July 04, 2017
  The Golgi apparatus (GA) is a ribbon‐like system of stacks which consist of multiple closely apposed flattened cisternae and vesicles usually localized in the juxta‐nuclear area. As for the biological functions, the GA plays a major role in protein biosynthesis, post‐translational modification, and sorting protein from ER to plasma membrane and other destinations. Structural changes and functional disorder of the GA is associated with various diseases. Moreover, increasing evidence revealed that swelling, poor development, and other morphological alterations of the GA are linked to cardiovascular diseases such as heart failure (HF), arrhythmia, and dilated cardiomyopathy. Furthermore, dysfunction of the GA is also related to cardiovascular diseases since the GA is extremely responsible for transport, glycosylation, biosynthesis, and subcellular distribution of cardiovascular proteins. This review gives a brief overview of the intricate relationship between the GA and cardiovascular diseases. In addition, we provide a further prospective that the GA may provide diagnosis reference for cardiovascular diseases, and changes in the ultrastructure and morphology of the GA such as swelling, poor development, and fragmentation may serve as a reliable index for cardiovascular diseases. The GA presents evident changes in the structure, morphology, and polarity in different cardiovascular diseases. The GA is responsible for transport, glycosylation, biosynthesis, and subcellular distribution of proteins which have biological effects on cardiovascular system.

  July 04, 2017   doi: 10.1002/jcp.26039   open full text
• Roles of macrophage migration inhibitory factor in cartilage tissue engineering.
  Yuko Fujihara, Atsuhiko Hikita, Tsuyoshi Takato, Kazuto Hoshi.
  Journal of Cellular Physiology. July 04, 2017
  To obtain stable outcomes in regenerative medicine, understanding and controlling immunological responses in transplanted tissues are of great importance. In our previous study, auricular chondrocytes in tissue‐engineered cartilage transplanted in mice were shown to express immunological factors, including macrophage migration inhibitory factor (MIF). Since MIF exerts pleiotropic functions, in this study, we examined the roles of MIF in cartilage regenerative medicine. We made tissue‐engineered cartilage consisting of auricular chondrocytes of C57BL/6J mouse, atellocollagen gel and a PLLA scaffold, and transplanted the construct subcutaneously in a syngeneic manner. Localization of MIF was prominent in cartilage areas of tissue‐engineered cartilage at 2 weeks after transplantation, though it became less apparent by 8 weeks. Co‐culture with RAW264 significantly increased the expression of MIF in chondrocytes, suggesting that the transplanted chondrocytes in tissue‐engineered cartilage could enhance the expression of MIF by stimulation of surrounding macrophages. When MIF was added in the culture of chondrocytes, the expression of type II collagen was increased, indicating that MIF could promote the maturation of chondrocytes. Meanwhile, toluidine blue staining of constructs containing wild type (Mif+/+) chondrocytes showed increased metachromasia compared to MIF‐knockout (Mif−/−) constructs at 2 weeks. However, this tendency was reversed by 8 weeks, suggesting that the initial increase in cartilage maturation in Mif+/+ constructs deteriorated by 8 weeks. Since the Mif+/+ constructs included more iNOS‐positive inflammatory macrophages at 2 weeks, MIF might induce an M1 macrophage‐polarized environment, which may eventually worsen the maturation of tissue‐engineered cartilage in the long term. In the present study, we examined the roles of MIF in cartilage regenerative medicine. MIF increased the expression of type II collagen in chondrocytes in an autocrine manner, promoting the maturation of tissue‐engineered cartilage. Meanwhile, MIF in chondrocytes could also regulate the phenotype of surrounding macrophages toward an inflammatory state, exacerbating the maturation of transplanted tissues in the long term.

  July 04, 2017   doi: 10.1002/jcp.26036   open full text
• G protein‐coupled receptor 84 controls osteoclastogenesis through inhibition of NF‐κB and MAPK signaling pathways.
  Ji‐Wan Park, Hye‐Jin Yoon, Woo Youl Kang, Seungil Cho, Sook Jin Seong, Hae Won Lee, Young‐Ran Yoon, Hyun‐Ju Kim.
  Journal of Cellular Physiology. July 04, 2017
  GPR84, a member of the G protein‐coupled receptor family, is found predominantly in immune cells, such as macrophages, and functions as a pivotal modulator of inflammatory responses. In this study, we investigated the role of GPR84 in receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclast differentiation. Our microarray data showed that GPR84 was significantly downregulated in osteoclasts compared to in their precursors, macrophages. The overexpression of GPR84 in bone marrow‐derived macrophages suppressed the formation of multinucleated osteoclasts without affecting precursor proliferation. In addition, GPR84 overexpression attenuated the induction of c‐Fos and nuclear factor of activated T cells, cytoplasmic 1 (NFATc1), which are transcription factors that are critical for osteoclastogenesis. Furthermore, knockdown of GPR84 using a small hairpin RNA promoted RANKL‐mediated osteoclast differentiation and gene expression of osteoclastogenic markers. Mechanistically, GPR84 overexpression blocked RANKL‐stimulated phosphorylation of IκBα and three MAPKs, JNK, ERK, and p38. GPR84 also suppressed NF‐κB transcriptional activity mediated by RANKL. Conversely, GPR84 knockdown enhanced RANKL‐induced activation of IκBα and the three MAPKs. Collectively, our results revealed that GPR84 functions as a negative regulator of osteoclastogenesis, suggesting that it may be a potential therapeutic target for osteoclast‐mediated bone‐destructive diseases. GPR84 inhibits RANKL‐induced NF‐κB and MAPKs signaling pathways and subsequently attenuates the expression of c‐Fos and NFATc1, which are key transcription factors during early osteoclastogenesis. This leads to down‐regulation of osteoclastogenic gene expression, thereby ultimately suppressing osteoclast differentiation.

  July 04, 2017   doi: 10.1002/jcp.26035   open full text
• Zebrafish Tmem230a cooperates with the Delta/Notch signaling pathway to modulate endothelial cell number in angiogenic vessels.
  Silvia Carra, Lorenzo Sangiorgio, Paride Pelucchi, Solei Cermenati, Alessandra Mezzelani, Valentina Martino, Mira Palizban, Alberto Albertini, Martin Götte, James Kehler, Gianluca Deflorian, Monica Beltrame, Antonio Giordano, Rolland Reinbold, Franco Cotelli, Gianfranco Bellipanni, Ileana Zucchi.
  Journal of Cellular Physiology. July 04, 2017
  During embryonic development, new arteries, and veins form from preexisting vessels in response to specific angiogenic signals. Angiogenic signaling is complex since not all endothelial cells exposed to angiogenic signals respond equally. Some cells will be selected to become tip cells and acquire migration and proliferation capacity necessary for vessel growth while others, the stalk cells become trailer cells that stay connected with pre‐existing vessels and act as a linkage to new forming vessels. Additionally, stalk and tip cells have the capacity to interchange their roles. Stalk and tip cellular responses are mediated in part by the interactions of components of the Delta/Notch and Vegf signaling pathways. We have identified in zebrafish, that the transmembrane protein Tmem230a is a novel regulator of angiogenesis by its capacity to regulate the number of the endothelial cells in intersegmental vessels by co‐operating with the Delta/Notch signaling pathway. Modulation of Tmem230a expression by itself is sufficient to rescue improper number of endothelial cells induced by aberrant expression or inhibition of the activity of genes associated with the Dll4/Notch pathway in zebrafish. Therefore, Tmem230a may have a modulatory role in vessel‐network formation and growth. As the Tmem230 sequence is conserved in human, Tmem230 may represent a promising novel target for drug discovery and for disease therapy and regenerative medicine in promoting or restricting angiogenesis. We have identified in zebrafish, that the transmembrane protein Tmem230a is a novel regulator of angiogenesis by its capacity to regulate the number of the endothelial cells in intersegmental vessels by co‐operating with the Delta/Notch signaling pathway. Modulation of Tmem230a expression by itself is sufficient to rescue improper number of endothelial cells induced by aberrant expression or inhibition of the activity of genes associated with the Dll4/Notch pathway in zebrafish. As the Tmem230 sequence is conserved in human, tmem230 may represent a promising novel target for drug discovery and for disease therapy and regenerative medicine in promoting or restricting angiogenesis.

  July 04, 2017   doi: 10.1002/jcp.26032   open full text
• The genetic factors contributing to the development of Wilm's tumor and their clinical utility in its diagnosis and prognosis.
  Afsane Bahrami, Marjan Joodi, Mina Maftooh, Gordon A. Ferns, Mehrdad M. Ahmadi, Seyed M. Hassanian, Amir Avan.
  Journal of Cellular Physiology. July 04, 2017
  Mutations in the Wilm's tumor 1 (WT1) gene are associated with a wide spectrum of renal manifestations, ultimately leading to end‐stage kidney failure. There is an inadequate understanding of the molecular functions of WT1 in renal development, and this has limited the potential for therapeutic interventions in WT1‐related diseases. In this review, we discuss the existing data on the genetic and epigenetic abnormalities that have been described in WTs and their potential utility as biomarkers for risk stratification, prediction and prognosis in patients with WTs.

  July 04, 2017   doi: 10.1002/jcp.26021   open full text
• Transcription‐Dependent Association of HDAC2 with Active Chromatin.
  Sanzida Jahan, Jian‐Min Sun, Shihua He, James Ronald Davie.
  Journal of Cellular Physiology. July 03, 2017
  Histone deacetylase 2 (HDAC2) catalyzes deacetylation of histones at the promoter and coding regions of transcribed genes and regulates chromatin structure and transcription. To explore the role of HDAC2 and phosphorylated HDAC2 in gene regulation, we studied the location along transcribed genes, the mode of recruitment and the associated proteins with HDAC2 and HDAC2S394ph in chicken polychromatic erythrocytes. We show that HDAC2 and HDAC2S394ph are associated with transcriptionally active chromatin and located in the interchromatin channels. HDAC2S394ph was present primarly at the upstream promoter region of the transcribed CA2 and GAS41 genes, while total HDAC2 was also found within the coding region of the CA2 gene. Recruitment of HDAC2 to these genes was partially dependent upon on‐going transcription. Unmodified HDAC2 was associated with RNA binding proteins and interacted with RNA bound to the initiating and elongating forms of RNA polymerase II. HDAC2S394ph was not associated with RNA polymerase II. These results highlight the differential properties of unmodified and phosphorylated HDAC2 and the organization of acetylated transcriptionally active chromatin in the chicken polychromatic erythrocyte. This article is protected by copyright. All rights reserved

  July 03, 2017   doi: 10.1002/jcp.26078   open full text
• Src nuclear localization and its prognostic relevance in human osteosarcoma.
  Enrica Urciuoli, Ilenia Coletta, Emanuele Rizzuto, Rita De Vito, Stefania Petrini, Valentina D'Oria, Marco Pezzullo, Giuseppe Maria Milano, Raffaele Cozza, Franco Locatelli, Barbara Peruzzi.
  Journal of Cellular Physiology. July 03, 2017
  Osteosarcoma is the most common malignant bone tumor in children and young adults. The identification of proteins which exhibit different subcellular localization in low‐ versus high‐risk osteosarcoma can be instrumental to obtain prognostic information and to develop innovative therapeutic strategies. Beside the well‐characterised membrane and cytoplasmic localization of Src protein, this study evaluated the prognostic relevance of its so‐far unknown nuclear compartmentalization. We analyzed the subcellular distribution of total and activated (pY418) Src in a tissue microarray including 60 osteosarcoma samples. Immunohistochemical analyses revealed a variable pattern of Src expression and localization, ranging from negative to high‐stained nuclei combined with a substantial cytoplasmic staining for total and activated forms. The analysis of Kaplan‐Meier survival curves in relationship to the diverse permutations of cytoplasmic and nuclear staining suggested a correlation between Src subcellular localization and the overall survival of osteosarcoma patients. In order to explain this different subcellular localization, normal osteoblasts and three osteosarcoma cell lines were used to investigate the molecular mechanism. Once confirmed a variable Src localization also in these cell lines, we demonstrated a correlation between the N‐myristoyltransferase enzymes expression and activity and the Src nuclear content. In conclusion, these results described a so‐far unknown Src nuclear localization in osteosarcoma cells, suggesting that the combined detection of nuclear and cytoplasmic Src levels can be used as a prognostic marker for osteosarcoma patient survival. A correlation between the N‐myristoyltransferase enzymes and the Src subcellular localization was described as well. This article is protected by copyright. All rights reserved

  July 03, 2017   doi: 10.1002/jcp.26079   open full text
• TGF‐β and Th17 Cells Related Injuries in Patients with Sulfur Mustard Exposure.
  Yunes Panahi, Mostafa Ghanei, Saeed Hassani, Amirhossein Sahebkar.
  Journal of Cellular Physiology. July 01, 2017
  Sulfur mustard (SM) is a vesicating agent that has been employed as a chemical warfare agent. High‐dose exposure to sulfur mustard may lead to the damage of rapidly proliferating cells of bone marrow and, therefore, suppression of the immune system. This may be continued as dysfunction of the immune system, and ultimately result in secondary immune disorders. Studies have suggested a role for T cells in SM‐induced lung injury. Moreover, observations from animal studies indicate a delayed‐type hypersensitivity (DTH) response after skin exposure to SM, providing an understanding that SM can stimulate specific T cell‐mediated immune responses. On the other hand, T helper (Th) 17 cells, which are a subset of CD4+ T cells, have recently been reported to be involved in a number of inflammatory, autoimmune, and chronic fibrotic lung diseases. Furthermore, a strong association has been established between the overproduction of profibrotic cytokines like transforming growth factor (TGF)‐β and Th17 cell number. In this review, we aimed to go through the new findings about the involvement and interactions of TGF‐β and Th17 in SM‐related injuries. This article is protected by copyright. All rights reserved

  July 01, 2017   doi: 10.1002/jcp.26077   open full text
• Targeting the tumor microenvironment as a potential therapeutic approach in colorectal cancer: Rational and progress.
  Afsane Bahrami, Majid Khazaei, Seyed Mahdi Hassanian, Soodabeh ShahidSales, Mona Joudi‐Mashhad, Mina Maftouh, Mir Hadi Jazayeri, Mohammad Reza Parizade, Gordon A. Ferns, Amir Avan.
  Journal of Cellular Physiology. June 30, 2017
  Colorectal cancer (CC) is often diagnosed at a late stage when tumor metastasis may have already occurred. Current treatments are often ineffective in metastatic disease, and consequently late diagnosis is often associated with poor outcomes in CC. Alternative strategies are therefore urgently required. An interaction between epithelial cancer cells and their tissue microenvironment is a contributor to metastasis, and therefore recent studies are beginning to focus on the properties of the tumor microenvironment and the mechanism by which the metastatic cells exploit the tumor microenvironment for survival, immune evasion, and growth. We have reviewed the development of the combined therapeutic approaches that have focused on targeting the microenvironment of CC.

  June 30, 2017   doi: 10.1002/jcp.26041   open full text
• Myeloid‐derived Suppressor Cells: important contributors to tumor progression and metastasis.
  Elham Safarzadeh, Mona Orangi, Hamed Mohammadi, Farhad Babai, Behzad Baradaran.
  Journal of Cellular Physiology. June 29, 2017
  Myeloid‐derived suppressor cells (MDSCs) are traditionally considered among the major components of the immunosuppressive tumor microenvironment. However, there is currently increasing evidence indicating that MDSCs in addition to suppression of immune surveillance is also involved in an array of non‐immunological functions like augmenting metastatic potential of tumor cells. Indeed, MDSCs can promote metastasis in animal models and cancer patients through promoting premetastatic niche formation, tumor angiogenesis and invasion. Moreover, MDSC frequency and function have been associated with progressive disease and correlated with clinical outcome. This review will summarize and discusses the data demonstrating the role for MDSCs in tumor metastasis. This article is protected by copyright. All rights reserved

  June 29, 2017   doi: 10.1002/jcp.26075   open full text
• Silibinin Stimluates Apoptosis by Inducing Generation of ROS and ER Stress in Human Choriocarcinoma Cells.
  Jiyeon Ham, Whasun Lim, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. June 28, 2017
  Silibinin is a flavonolignan extracted from seeds of milk thistles. Traditionally, it has been used as a therapeutic agent for liver disorders, and now it is well‐known for its anti‐cancer effects. However, studies on anti‐cancer effects of silibinin on choriocarcinoma are very limited. Therefore, we performed proliferation and apoptosis assays to determine effects of silibinin on the viability of human choriocarcinoma (JAR and JEG3) cells. Our results showed that silibinin significantly inhibited proliferation and induced apoptosis in both JAR and JEG3 cells, and significantly increased reactive oxygen species (ROS) and lipid peroxidation. Moreover, silibinin disrupted mitochondrial function by inducing permeabilization of mitochondrial membrane potential and calcium ion efflux in JAR and JEG3 cells. Furthermore, silibinin‐induced apoptosis in choriocarcinoma cells via AKT, mitogen‐activated protein kinases (MAPK) and unfolded protein response (UPR) signal transduction. Collectively, our results suggest that silibinin is a novel therapeutic agent or dietary supplement for management of human placental choriocarcinomas. This article is protected by copyright. All rights reserved

  June 28, 2017   doi: 10.1002/jcp.26069   open full text
• Ptn functions downstream of C/EBPβ to mediate the effects of cAMP on uterine stromal cell differentiation through targeting Hand2 in response to progesterone.
  Hai‐Fan Yu, Ran Tao, Zhan‐Qing Yang, Kai Wang, Zhan‐Peng Yue, Bin Guo.
  Journal of Cellular Physiology. June 28, 2017
  Ptn is a pleiotropic growth factor involving in the regulation of cellular proliferation and differentiation, but its biological function in uterine decidualization remains unknown. Here we showed that Ptn was highly expressed in the decidual cells, and could induce the proliferation of uterine stromal cells and expression of Prl8a2 and Prl3c1 which were two well‐established differentiation markers for decidualization, suggesting an important role of Ptn in decidualization. In the uterine stromal cells, progesterone stimulated the expression of Ptn accompanied with an accumulation of intracellular cAMP level. Silencing of Ptn impeded the induction of progesterone and cAMP on the differentiation of uterine stromal cells. Administration of PKA inhibitor H89 resulted in a blockage of progesterone on Ptn expression. Further analysis evidenced that regulation of progesterone and cAMP on Ptn was mediated by C/EBPβ. During in vitro decidualization, knockdown of Ptn could weaken the up‐regulation of Prl8a2 and Prl3c1 elicited by C/EBPβ overexpression, while constitutive activation of Ptn reversed the repressive effects of C/EBPβ siRNA on the expression of Prl8a2 and Prl3c1. Meanwhile, Ptn might mediate the regulation of C/EBPβ on Hand2 which was a downstream target of Ptn in the differentiation of uterine stromal cells. Attenuation of Ptn or C/EBPβ by specific siRNA blocked the stimulation of Hand2 by progesterone and cAMP. Collectively, Ptn may play a vital role in the progesterone‐induced decidualization pathway. This article is protected by copyright. All rights reserved

  June 28, 2017   doi: 10.1002/jcp.26067   open full text
• Reprogramming mouse embryo fibroblasts to functional islets without genetic manipulation.
  Bhawna Chandravanshi, Ramesh Bhonde.
  Journal of Cellular Physiology. June 28, 2017
  The constant quest for generation of large number of islets aimed us to explore the differentiation potential of mouse embryo fibroblast cells. Mouse embryo fibroblast cells isolated from 12‐14 day old pregnant mice were characterized for their surface markers and tri‐lineage differentiation potential. They were subjected to serum free media containing a cocktail of islet differentiating reagents and analysed for the expression of pancreatic lineage transcripts .The islet like cell aggregates (ICAs) was confirmed for their pancreatic properties via immunofluorecence for C‐peptide, glucagon and somatostain. They were positive for CD markers‐ Sca1, CD44, CD73 and CD90 and negative for haematopoietic markers‐ CD34 and CD45 at both transcription and translational levels. The transcriptional analysis of the ICAs at different day points exhibited up‐regulation of islet markers (Insulin, PDX1, HNF3, Glucagon and Somatostatin) and down‐regulation of MSC‐ markers (Vimentin and Nestin). They positively stained for dithizone, C‐peptide, insulin, glucagon and somatostatin indicating intact insulin producing machinery. In‐vitro glucose stimulation assay revealed 3 fold increase in insulin secretion as compared to basal glucose with insulin content being the same in both the conditions. The preliminary in vivo data on ICA transplantation showed reversal of diabetes in streptozotocin induced diabetic mice. Our results demonstrate for the first time that mouse embryo fibroblast cells contain a population of MSC like cells which could differentiate into insulin producing cell aggregates. Hence our study could be extrapolated for isolation of MSC like cells from human, medically terminated pregnancies to generate ICAs for treating type 1 diabetic patients. This article is protected by copyright. All rights reserved

  June 28, 2017   doi: 10.1002/jcp.26068   open full text
• LncRNA‐PVT1 promotes pancreatic cancer cells proliferation and migration through acting as a molecular sponge to regulate miR‐448.
  Liang Zhao, Hongru Kong, Hongwei Sun, Zongjing Chen, Bicheng Chen, Mengtao Zhou.
  Journal of Cellular Physiology. June 28, 2017
  The identification and characterization of long non‐coding RNAs (lncRNAs) in diverse biological process has currently developed rapidly. LncRNA‐PVT1, located adjacent to the MYC locus on chromosomal region 8q24, has been reported to be associated with many biological processes. However, the function and mechanism of PVT1 in pancreatic carcinoma (PC) is poorly understood. In this present study, we first measured the level of PVT1 in the PC cell lines and tissues by quantitative real‐time PCR (qRT‐PCR), and then employed loss‐of‐function and gain‐of‐function approaches to explore the association between PVT1 expression levels and PC cell proliferation/migration ability. Furthermore, bioinformatics analysis was utilized to show that PVT1 contains binding site for miR‐448 and an inverse correlation between PVT1 and miR‐448 was obtained in PC specimens. Additionally, dual luciferase reporter assay, RNA‐binding protein immunoprecipitation (RIP) and applied biotin‐avidin pulldown system were applied to further confirm that PVT1 directly bind with microRNA binding site harboring in the PVT1 sequence. Then, SERBP1 was identified as a target of miR‐448 according to the gene expression array analysis of PC clinical samples. Together, we revealed that PVT1 functions as an endogenous ‘sponge’ by competing for miR‐448 binding to regulate the miRNA target SERBP1 and therefore promotes the proliferation and migration of PC cells. This article is protected by copyright. All rights reserved

  June 28, 2017   doi: 10.1002/jcp.26072   open full text
• Hypoxia‐induced apoptosis of mouse spermatocytes is mediated by HIF‐1α through a death receptor pathway and a mitochondrial pathway.
  Jun Yin, Bing Ni, Wei‐Gong Liao, Yu‐Qi Gao.
  Journal of Cellular Physiology. June 27, 2017
  Hypoxia in vivo induces oligozoospermia, azoospermia, and degeneration of the germinal epithelium, but the underlying molecular mechanism of this induction is not fully clarified. The aim of this study was to investigate the role of the death receptor pathway and the mitochondrial pathway in hypoxia‐induced apoptosis of mouse GC‐2spd (GC‐2) cells and the relationship between HIF‐1α and apoptosis of GC‐2 cells induced by hypoxia. GC‐2 cells were subjected to 1% oxygen for 48 hr. Apoptosis was detected by flow cytometry, TUNEL staining, LDH, caspase‐3/8/9 in the absence and presence of HIF‐1α siRNA. The protein levels of apoptosis‐related markers were determined by Western blot in the presence and absence of HIF‐1α siRNA. Mitochondrial transmembrane potential change was observed by in situ JC‐1 staining. Cell viability was assessed upon treatment of caspase‐8 and 9 inhibitors. The results indicated that hypoxia at 1% oxygen for 48 hr induced apoptosis of GC‐2 cells. A prolonged exposure of GC‐2 cells to hypoxic conditions caused downregulation of c‐FLIP, DcR2 and Bcl‐2 and upregulation of DR5, TRAIL, Fas, p53, and Bax, with an overproduction of caspase‐3/8/9. Moreover, hypoxia at this level had an effect on mitochondrial depolarization. In addition, specific inhibitors of caspase‐8/9 partially suppressed hypoxia‐induced GC‐2 cell apoptosis, and the anti‐apoptotic effects of the caspase inhibitors were additive. Of note, HIF‐1α knockdown attenuated hypoxia and induced apoptosis of GC‐2 cells. In conclusion, our data suggest that the death receptor pathway and mitochondrial pathway, which are likely mediated by HIF‐1α, contribute to hypoxia‐induced GC‐2 cell apoptosis. It has been known that hypoxia induces male infertility with the underlying mechanisms to be clarified. This study determined that the death receptor pathway and mitochondrial pathway, which are likely mediated by HIF‐1α, contribute to hypoxia‐induced apoptosis of mouse pachytene spermatocytes.

  June 27, 2017   doi: 10.1002/jcp.25974   open full text
• Role of follistatin in muscle and bone alterations induced by gravity change in mice.
  Naoyuki Kawao, Hironobu Morita, Koji Obata, Kohei Tatsumi, Hiroshi Kaji.
  Journal of Cellular Physiology. June 22, 2017
  Interactions between muscle and bone have been recently noted. We reported that the vestibular system plays crucial roles in the changes in muscle and bone induced by hypergravity in mice. However, the details of the mechanisms by which gravity change affects muscle and bone through the vestibular system still remain unknown. Here, we investigated the roles of humoral factors linking muscle to bone and myostatin‐related factors in the hypergravity‐induced changes in muscle and bone in mice with vestibular lesions (VL). Hypergravity elevated serum and mRNA levels of follistatin, an endogenous inhibitor of myostatin, in the soleus muscle of mice. VL blunted the hypergravity‐enhanced levels of follistatin in the soleus muscle of mice. Simulated microgravity decreased follistatin mRNA level in mouse myoblastic C2C12 cells. Follistatin elevated the mRNA levels of myogenic genes as well as the phosphorylation of Akt and p70S6 kinase in C2C12 cells. As for bone metabolism, follistatin antagonized the mRNA levels of osteogenic genes suppressed by activin A during the differentiation of mesenchymal cells into osteoblastic cells. Moreover, follistatin attenuated osteoclast formation enhanced by myostatin in the presence of receptor activator of nuclear factor‐κB ligand in RAW 264.7 cells. Serum follistatin levels were positively related to bone mass in mouse tibia. In conclusion, the present study provides novel evidence that hypergravity affects follistatin levels in muscle through the vestibular system in mice. Follistatin may play some roles in the interactions between muscle and bone metabolism in response to gravity change. We provide novel evidence that hypergravity affects follistatin levels in muscle through the vestibular system in mice. The present study suggests that follistatin plays some roles by both direct action on muscle and action on bone as a circulating myokine in the regulation of muscle and bone in response to gravity changes.

  June 22, 2017   doi: 10.1002/jcp.25986   open full text
• MicroRNA: Relevance to stroke diagnosis, prognosis, and therapy.
  Hamed Mirzaei, Fatemeh Momeni, Leila Saadatpour, Amirhossein Sahebkar, Mohammad Goodarzi, Aria Masoudifar, Shirin Kouhpayeh, Hossein Salehi, Hamid Reza Mirzaei, Mahmoud Reza Jaafari.
  Journal of Cellular Physiology. June 22, 2017
  Stroke is a life‐threatening disease that accounts for a considerable burden of mortality in both developing and developed world. Identification of specific biomarkers for stroke and its outcomes can greatly contribute to improved care of patients. MicroRNAs (miRNAs) are known as novel biomarkers that could be used as diagnostic, prognostic, and therapeutic biomarkers. Various studies have shown that miRNAs have key roles in the pathogenesis of stroke, and its complications and outcomes. In addition, there is evidence showing that mesenchaymal stromal cell‐derived exosomes containing miRNAs can be used for monitoring and treatment of various diseases such as stroke. Here, we summarized various aspects of miRNA applications in different stages of stroke.

  June 22, 2017   doi: 10.1002/jcp.25787   open full text
• Therapeutic application of multipotent stem cells.
  Hamed Mirzaei, Amirhossein Sahebkar, Laleh Shiri Sichani, Abdullah Moridikia, Sara Nazari, Javid Sadri Nahand, Hossein salehi, Jan Stenvang, Aria Masoudifar, Hamid R. Mirzaei, Mahmoud R. Jaafari.
  Journal of Cellular Physiology. June 22, 2017
  Cell therapy is an emerging fields in the treatment of various diseases such as cardiovascular, pulmonary, hepatic, and neoplastic diseases. Stem cells are an integral tool for cell therapy. Multipotent stem cells are an important class of stem cells which have the ability to self‐renew through dividing and developing into multiple specific cell types in a specific tissue or organ. These cells are capable to activate or inhibit a sequence of cellular and molecular pathways leading to anti‐inflammatory and anti‐apoptotic effects which might contribute to the treatment of various diseases. It has been showed that multipotent stem cells exert their therapeutic effects via inhibition/activation of a sequence of cellular and molecular pathways. Although the advantages of multipotent stem cells are numerous, further investigation is still necessary to clarify the biology and safety of these cells before they could be considered as a potential treatment for different types of diseases. This review summarizes different features of multipotent stem cells including isolation, differentiation, and therapeutic applications.

  June 22, 2017   doi: 10.1002/jcp.25990   open full text
• S‐Adenosylmethionine‐mediated apoptosis is potentiated by autophagy inhibition induced by chloroquine in human breast cancer cells.
  Donatella Delle Cave, Vincenzo Desiderio, Laura Mosca, Concetta P. Ilisso, Luigi Mele, Michele Caraglia, Giovanna Cacciapuoti, Marina Porcelli.
  Journal of Cellular Physiology. June 22, 2017
  The naturally occurring sulfonium compound S‐adenosyl‐L‐methionine (AdoMet) is an ubiquitous sulfur‐nucleoside that represents the main methyl donor in numerous methylation reactions. In recent years, it has been shown that AdoMet possesses antiproliferative properties in various cancer cells, but the molecular mechanisms at the basis of the effect induced by AdoMet have been only in part investigated. In the present study, we found that AdoMet strongly inhibited the proliferation of breast cancer cells MCF‐7 by inducing both autophagy and apoptosis. AdoMet consistently enhanced the levels of the autophagy markers beclin‐1 and LC3B‐II, and caused a significant increase of pro‐apoptotic Bax/Bcl‐2 ratio paralleled by poly (ADP ribose) polymerase (PARP) and caspase 9, and 6 cleavage. Notably, AdoMet, already at low doses, raised the percentage of cells in G2/M phase of cell cycle by down‐regulating the expression of cell cycle‐regulatory proteins cyclin B and cyclin E with a remarkable increase of p53, p27, and p21. We also evaluated the combination of AdoMet and the autophagy inhibitor chloroquine (CLC) showing that autophagy block is synergistic in inducing both growth inhibition and apoptosis. These effects were paralleled by a strong inhibition of the activity of AKT and of the downstream effector mTOR and by an increased cleavage of caspase‐6 and PARP. These data suggest, for the first time, that autophagy can act as an escape mechanism from the apoptotic activity of AdoMet, and that AdoMet could be used in combination with CLC or its analogs in the treatment of breast cancer. AdoMet induces apoptosis in breast cancer cells through the inhibition of Akt. Autophagy is also activated and is a mechanism of escape from apoptosis induced by AdoMet. Cloroquine and AdoMet combination strongly potentiates the apoptosis induced by AdoMet blocking autophagy occurrence.

  June 22, 2017   doi: 10.1002/jcp.26015   open full text
• Differentiation of human olfactory bulb‐derived neural stem cells toward oligodendrocyte.
  Hany E. Marei, Zeinab Shouman, Asma Althani, Nahla Afifi, Abd‐Elmaksoud A, Samah Lashen, Anwarul Hasan, Thomas Caceci, Roberto Rizzi, Carlo Cenciarelli, Patrizia Casalbore.
  Journal of Cellular Physiology. June 22, 2017
  In the central nervous system (CNS), oligodendrocytes are the glial element in charge of myelin formation. Obtaining an overall presence of oligodendrocyte precursor cells/oligodendrocytes (OPCs/OLs) in culture from different sources of NSCs is an important research area, because OPCs/OLs may provide a promising therapeutic strategy for diseases affecting myelination of axons. The present study was designed to differentiate human olfactory bulb NSCs (OBNSCs) into OPCs/OLs and using expression profiling (RT‐qPCR) gene, immunocytochemistry, and specific protein expression to highlight molecular mechanism(s) underlying differentiation of human OBNSCs into OPCs/OLs. The differentiation of OBNSCs was characterized by a simultaneous appearance of neurons and glial cells. The differentiation medium, containing cAMP, PDGFA, T3, and all‐trans‐retinoic acid (ATRA), promotes OBNSCs to generate mostly oligodendrocytes (OLs) displaying morphological changes, and appearance of long cytoplasmic processes. OBNSCs showed, after 5 days in OLs differentiation medium, a considerable decrease in the number of nestin positive cells, which was associated with a concomitant increase of NG2 immunoreactive cells and few O4(+)‐OPCs. In addition, a significant up regulation in gene and protein expression profile of stage specific cell markers for OPCs/OLs (CNPase, Galc, NG2, MOG, OLIG1, OLIG2, MBP), neurons, and astrocytes (MAP2, β‐TubulinIII, GFAP) and concomitant decrease of OBNSCs pluripotency markers (Oct4, Sox2, Nestin), was demonstrated following induction of OBNSCs differentiation. Taken together, the present study demonstrate the marked ability of a cocktail of factors containing PDGFA, T3, cAMP, and ATRA, to induce OBNSCs differentiation into OPCs/OLs and shed light on the key genes and pathological pathways involved in this process. The present study demonstrate the marked ability of a cocktail of factors containing PDGFA, T3, cAMP, and ATRA, to induce OBNSCs differentiation into OPCs/OLs and shed light on the key genes and pathological pathways involved in this process.

  June 22, 2017   doi: 10.1002/jcp.26008   open full text
• MSCs ameliorates DPN induced cellular pathology via [Ca2+]i homeostasis and scavenging the pro‐inflammatory cytokines.
  Harish C. Chandramoorthy, Ismaeel Bin‐Jaliah, Hussian Karari, Prasanna Rajagopalan, Mohammed Eajaz Ahmed Shariff, Ahmed Al‐Hakami, Suliman M. Al‐Humayad, Fawzi A. Baptain, Humeda Suekit Ahmed, Hanaa Z. Yassin, Mohamed A. Haidara.
  Journal of Cellular Physiology. June 22, 2017
  The MSCs of various origins are known to ameliorate or modulate cell survival strategies. We investigated, whether UCB MSCs could improve the survival of the human neuronal cells and/or fibroblast assaulted with DPN sera. The results showed, the co‐culture of UCB MSCs with human neuronal cells and/or fibroblasts could effectively scavenge the pro‐inflammatory cytokines TNF‐α, IL‐1β, IFN‐ɤ and IL − 12 and control the pro‐apoptotic expression of p53/Bax. Further co‐culture of UCB MSCs have shown to induce anti‐inflammatory cytokines like IL‐4, IL‐10 and TGF‐β and anti‐apoptotic Bclxl/Bcl2 expression in the DPN sera stressed cells. Amelioration of elevated [Ca2+]i and cROS, the portent behind the NFκB/Caspase‐3 mediated inflammation in DPN rescued the cells from apoptosis. The results of systemic administration of BM MSCs improved DPN pathology in rat as extrapolated from human cell model. The BM MSCs ameliorated prolonged distal motor latency (control: 0.70 ± 0.06, DPN: 1.29 ± 0.13 m/s DPN + BM MSCs: 0.89 ± 0.02 m/s, p < 0.05) and lowered high amplitude of compound muscle action potentials (CMAPs) (control: 12.36 ± 0.41, DPN: 7.52 ± 0.61 mV, DPN + MSCs: 8.79 ± 0.53 mV, p < 0.05), while slowly restoring the plasma glucose levels. Together, all these results showed that administration of BM or UCB MSCs improved the DPN via ameliorating pro‐inflammatory cytokine signaling and [Ca2+]i homeostasis. Mesenchymal stem cells ameliorate diabetic peripheral neuropathy through modulating the intracellular milieu of calcium/ROS mediated downstream apoptotic neuronal cell death.

  June 22, 2017   doi: 10.1002/jcp.26009   open full text
• The influence of rAAV2‐mediated SOX2 delivery into neonatal and adult human RPE cells; a comparative study.
  Razie Ezati, Azadeh Etemadzadeh, Zahra‐Soheila Soheili, Shahram Samiei, Ehsan Ranaei Pirmardan, Malihe Davari, Hoda Shams Najafabadi.
  Journal of Cellular Physiology. June 22, 2017
  Cell replacement is a promising therapy for degenerative diseases like age‐related macular degeneration (AMD). Since the human retina lacks regeneration capacity, much attention has been directed toward persuading for cells that can differentiate into retinal neurons. In this report, we have investigated reprogramming of the human RPE cells and concerned the effect of donor age on the cellular fate as a critical determinant in reprogramming competence. We evaluated the effect of SOX2 over‐expression in human neonatal and adult RPE cells in cultures. The coding region of human SOX2 gene was cloned into adeno‐associated virus (AAV2) and primary culture of human neonatal/adult RPE cells were infected by recombinant virus. De‐differentiation of RPE to neural/retinal progenitor cells was investigated by quantitative real‐time PCR and ICC for neural/retinal progenitor cells’ markers. Gene expression analysis showed 80‐fold and 12‐fold over‐expression for SOX2 gene in infected neonatal and adult hRPE cells, respectively. The fold of increase for Nestin in neonatal and adult hRPE cells was 3.8‐fold and 2.5‐fold, respectively. PAX6 expression was increased threefold and 2.5‐fold in neonatal/adult treated cultures. Howbeit, we could not detect rhodopsin, and CHX10 expression in neonatal hRPE cultures and expression of rhodopsin in adult hRPE cells. Results showed SOX2 induced human neonatal/adult RPE cells to de‐differentiate toward retinal progenitor cells. However, the increased number of PAX6, CHX10, Thy1, and rhodopsin positive cells in adult hRPE treated cultures clearly indicated the considerable generation of neuro‐retinal terminally differentiated cells.

  June 22, 2017   doi: 10.1002/jcp.25991   open full text
• ABC of multifaceted dystrophin glycoprotein complex (DGC).
  Hina F. Bhat, Saima S. Mir, Khalid B. Dar, Zuhaib F. Bhat, Riaz A. Shah, Nazir A. Ganai.
  Journal of Cellular Physiology. June 22, 2017
  Dystrophin protein in association with several other cellular proteins and glycoproteins leads to the formation of a large multifaceted protein complex at the cell membrane referred to as dystrophin glycoprotein complex (DGC), that serves distinct functions in cell signaling and maintaining the membrane stability as well as integrity. In accordance with this, several findings suggest exquisite role of DGC in signaling pathways associated with cell development and/or maintenance of homeostasis. In the present review, we summarize the established facts about the various components of this complex with emphasis on recent insights into specific contribution of the DGC in cell signaling at the membrane. We have also discussed the recent advances made in exploring the molecular associations of DGC components within the cells and the functional implications of these interactions. Our review would help to comprehend the composition, role, and functioning of DGC and may lead to a deeper understanding of its role in several human diseases. The dystrophin glycoprotein complex (DGC) is a membrane associated protein complex that plays a very crucial role in maintaining the integrity and stability of membranes. Growing evidence points toward a central role of DGC complex in membrane integrity, synapse, and crucial cell signaling pathways. Furthermore,a large number of muscular pathologies and synapse related disorders are being associated with mutations and/or deregulation of DGC signaling.

  June 22, 2017   doi: 10.1002/jcp.25982   open full text
• Intranuclear and higher‐order chromatin organization of the major histone gene cluster in breast cancer.
  Andrew J. Fritz, Prachi N. Ghule, Joseph R. Boyd, Coralee E. Tye, Natalie A. Page, Deli Hong, David J. Shirley, Adam S. Weinheimer, Ahmet R. Barutcu, Diana L. Gerrard, Seth Frietze, Andre J. van Wijnen, Sayyed K. Zaidi, Anthony N. Imbalzano, Jane B. Lian, Janet L. Stein, Gary S. Stein.
  Journal of Cellular Physiology. June 22, 2017
  Alterations in nuclear morphology are common in cancer progression. However, the degree to which gross morphological abnormalities translate into compromised higher‐order chromatin organization is poorly understood. To explore the functional links between gene expression and chromatin structure in breast cancer, we performed RNA‐seq gene expression analysis on the basal breast cancer progression model based on human MCF10A cells. Positional gene enrichment identified the major histone gene cluster at chromosome 6p22 as one of the most significantly upregulated (and not amplified) clusters of genes from the normal‐like MCF10A to premalignant MCF10AT1 and metastatic MCF10CA1a cells. This cluster is subdivided into three sub‐clusters of histone genes that are organized into hierarchical topologically associating domains (TADs). Interestingly, the sub‐clusters of histone genes are located at TAD boundaries and interact more frequently with each other than the regions in‐between them, suggesting that the histone sub‐clusters form an active chromatin hub. The anchor sites of loops within this hub are occupied by CTCF, a known chromatin organizer. These histone genes are transcribed and processed at a specific sub‐nuclear microenvironment termed the major histone locus body (HLB). While the overall chromatin structure of the major HLB is maintained across breast cancer progression, we detected alterations in its structure that may relate to gene expression. Importantly, breast tumor specimens also exhibit a coordinate pattern of upregulation across the major histone gene cluster. Our results provide a novel insight into the connection between the higher‐order chromatin organization of the major HLB and its regulation during breast cancer progression. Positional gene enrichment identified the major histone gene cluster at chromosome 6p22 as one of the most significantly upregulated (and not amplified) clusters of genes from the normal‐like MCF10A to premalignant MCF10AT1 and metastatic MCF10CA1a cells. This cluster is subdivided into three sub‐clusters of histone genes that are organized into hierarchical topologically associating domains (TADs) with the histone genes located at sub‐TAD boundaries. These sub‐clusters interact more frequently with each other than the regions in‐between them, suggesting that the histone sub‐clusters form an active chromatin hub.

  June 22, 2017   doi: 10.1002/jcp.25996   open full text
• Nanoparticles as new tools for inhibition of cancer angiogenesis.
  Nasser Hashemi Goradel, Farshid Ghiyami‐Hour, Samira Jahangiri, Babak Negahdari, Amirhossein Sahebkar, Aria Masoudifar, Hamed Mirzaei.
  Journal of Cellular Physiology. June 22, 2017
  Angiogenesis is known as one of the hallmarks of cancer. Multiple lines evidence indicated that vascular endothelium growth factor (VEGF) is a key player in the progression of angiogenesis and exerts its functions via interaction with tyrosine kinase receptors (TKRs). These receptors could trigger a variety of cascades that lead to the supply of oxygen and nutrients to tumor cells and survival of these cells. With respect to pivotal role of angiogenesis in the tumor growth and survival, finding new therapeutic approaches via targeting angiogenesis could open a new horizon in cancer therapy. Among various types of therapeutic strategies, nanotechnology has emerged as new approach for the treatment of various cancers. Nanoparticles (NPs) could be used as effective tools for targeting a variety of therapeutic agents. According to in vitro and in vivo studies, NPs are efficient in depriving tumor cells from nutrients and oxygen by inhibiting angiogenesis. However, the utilization of NPs are associated with a variety of limitations. It seems that new approaches such as NPs conjugated with hydrogels could overcome to some limitations. In the present review, we summarize various mechanisms involved in angiogenesis, common anti‐angiogenesis strategies, and application of NPs for targeting angiogenesis in various cancers.

  June 22, 2017   doi: 10.1002/jcp.26029   open full text
• Cholinergic and dopaminergic neuronal differentiation of human adipose tissue derived mesenchymal stem cells.
  Hany El Sayed Marei, Aya El‐Gamal, Asma Althani, Nahla Afifi, Ahmed Abd‐Elmaksoud, Amany Farag, Carlo Cenciarelli, Caceci Thomas, Hasan Anwarul.
  Journal of Cellular Physiology. June 22, 2017
  Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into various cell types such as cartilage, bone, and fat cells. Recent studies have shown that induction of MSCs in vitro by growth factors including epidermal growth factor (EGF) and fibroblast growth factor (FGF2) causes them to differentiate into neural like cells. These cultures also express ChAT, a cholinergic marker; and TH, a dopaminergic marker for neural cells. To establish a protocol with maximum differentiation potential, we examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP‐9, retinoic acid, and heparin. Adipose‐derived MSCs were extracted and expanded in vitro for 3 passages after reaching >80% confluency, for a total duration of 9 days. Cells were then characterized by flow cytometry for CD markers as CD44 positive and CD45 negative. MSCs were then treated with neural induction media and were characterized by morphological changes and Q‐PCR. Differentiated MSCs expressed markers for immature and mature neurons; β Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons. Improved protocols for MSCs induction will facilitate and ensure the reproducibility and standard production of MSCs for therapeutic applications in neurodegenerative diseases. We examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP‐9, retinoic acid, and heparin. MSCs were then treated with neural induction media and were characterized by morphological changes and Q‐PCR. Differentiated MSCs expressed markers for immature and mature neurons; β Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons.

  June 22, 2017   doi: 10.1002/jcp.25937   open full text
• ESRRB plays a crucial role in the promotion of porcine cell reprogramming.
  Fan Yang, Yahui Ren, Huan Li Huayan Wang.
  Journal of Cellular Physiology. June 21, 2017
  The estrogen‐related receptor b (ESRRB) is an orphan nuclear receptor and targets many genes involved in self‐renewal and pluripotency. In mouse ES cells, overexpression of ESRRB can maintain LIF‐independent self‐renewal in the absence of Nanog. However, the fundamental features of porcine ESRRB remain elusive. In this study, we revealed the expression profiles of ESRRB in both porcine pluripotent stem cells and early stage embryos and dissected the functional domains of ESRRB protein to prove that ESRRB is a key transcription factor that enhanced porcine pluripotent gene activation. Addition of ESRRB into the cocktail of core pluripotent factors Oct4, Sox2, Klf4, and c‐Myc (OSKM + E) could significantly enhance the reprogramming efficiency and the formation of alkaline phosphatase positive colonies. Conversely, knockdown of ESRRB in piPSCs significantly reduced the expression level of pluripotent genes, minimized the alkaline phosphatase activity, and initiated the porcine induced pluripotent stem cell differentiation. Therefore, porcine ESRRB is a crucial transcription factor to improve the self‐renewal of piPSCs. This article is protected by copyright. All rights reserved

  June 21, 2017   doi: 10.1002/jcp.26063   open full text
• The novel role of pyrvinium in cancer therapy.
  Amir A. Momtazi‐borojeni, Elham Abdollahi, Faezeh Ghasemi, Michele Caraglia, Amirhossein Sahebkar.
  Journal of Cellular Physiology. June 20, 2017
  Pyrvinium pamoate (PP) is a quinoline‐derived cyanine dye which was officially approved by FDA for its anthelmintic properties and therapeutic function against animal‐like protists such as Cryptosporidium parvum and Plasmodium falciparum in the 1950s. In the last 10 years, several studies have shown the novel activity of pyrvinium in tumor therapy. Some investigations have indicated that pyrvinium could delay or inhibit tumor cell proliferation in cancer models including colon, breast, lung and prostate cancer, and some hematological malignancies. In this review, we discuss multiple critical signaling pathways and mechanisms underlying the anticancer effects of PP. In details, pyrvinium acts through the following main mechanisms: (i) energy and autophagy depletion; and (ii) inhibition of Akt and Wnt‐β‐catenin‐dependent pathways. Interestingly, pyrvinium has also shown potent anti‐cancer stem cell activity. The overwhelming insights into the mechanism of anticancer properties of PP can help establishing novel and future anti‐tumor treatment strategies. This review presents the mechanisms of anticancer properties of pyrvinium that can help establishing novel and future anti‐tumor treatment strategies.

  June 20, 2017   doi: 10.1002/jcp.26006   open full text
• High fat diet attenuates hyperglycemia, body composition changes, and bone loss in male streptozotocin‐induced type 1 diabetic mice.
  Adriana Lelis Carvalho, Victoria E. DeMambro, Anyonya R. Guntur, Phuong Le, Kenichi Nagano, Roland Baron, Francisco José Albuquerque de Paula, Katherine J. Motyl.
  Journal of Cellular Physiology. June 20, 2017
  There is a growing and alarming prevalence of obesity and the metabolic syndrome in type I diabetic patients (T1DM), particularly in adolescence. In general, low bone mass, higher fracture risk and increased marrow adipose tissue (MAT) are features of diabetic osteopathy in insulin deficient subjects. On the other hand, type 2 diabetes (T2DM) is associated with normal or high bone mass, a greater risk of peripheral fractures and no change in MAT. Therefore, we sought to determine the effect of weight gain on bone turnover in insulin deficient mice. We evaluated the impact of a 6‐week high‐fat (HFD) rich in medium chain fatty acids or low‐fat diet (LFD) on bone mass and MAT in a streptozotocin (STZ)‐induced model using male C57BL/6J mice at 8 weeks of age. Dietary intervention was initiated after diabetes confirmation. At the endpoint, lower non‐fasting glucose levels were observed in diabetic mice fed with high fat diet compared to diabetic mice fed the low fat diet (STZ‐LFD). Compared to euglycemic controls, the STZ‐LFD had marked polydipsia and polyphagia, as well as reduced lean mass, fat mass and bone parameters. Interestingly, STZ‐HFD mice had higher bone mass, namely less cortical bone loss and more trabecular bone than STZ‐LFD. Thus, we found that a HFD, rich in medium chain fatty acids, protects against bone loss in a T1DM mouse model. Whether this may also translate to T1DM patients who are overweight or obese in respect to maintenance of bone mass remains to be determined through longitudinal studies. This article is protected by copyright. All rights reserved

  June 20, 2017   doi: 10.1002/jcp.26062   open full text
• High density lipoprotein (HDL) reverses palmitic acid induced energy metabolism imbalance by switching CD36 and GLUT4 signaling pathways in cardiomyocyte.
  Su‐Ying Wen, Bharath Kumar Velmurugan, Cecilia Hsuan Day, Chia‐Yao Shen, Li‐Chin Chun, Yi‐Chieh Tsai, Yueh‐Min Lin, Ray‐Jade Chen, Chia‐Hua Kuo, Chih‐Yang Huang.
  Journal of Cellular Physiology. June 16, 2017
  In our previous study palmitic acid (PA) induced lipotoxicity and switches energy metabolism from CD36 to GLUT4 in H9c2 cells. Low level of high density lipoprotein (HDL) is an independent risk factor for cardiac hypertrophy. Therefore, we in the present study investigated whether HDL can reverse PA induced lipotoxicity in H9c2 cardiomyoblast cells. In this study, we treated H9c2 cells with PA to create a hyperlipidemia model in vitro and analyzed for CD36 and GLUT4 metabolic pathway proteins. CD36 metabolic pathway proteins (phospho‐AMPK, SIRT1, PGC1α, PPARα, CPT1β, and CD36) were decreased by high PA (150 and 200 μg/μl) concentration. Interestingly, expression of GLUT4 metabolic pathway proteins (p‐PI3K and pAKT) were increased at low concentration (50 μg/μl) and decreased at high PA concentration. Whereas, phospho‐PKCζ, GLUT4 and PDH proteins expression was increased in a dose dependent manner. PA treated H9c2 cells were treated with HDL and analyzed for cell viability. Results showed that HDL treatment induced cell proliferation efficiency in PA treated cells. In addition, HDL reversed the metabolic effects of PA: CD36 translocation was increased and reduced GLUT4 translocation, but HDL treatment significantly increased CD36 metabolic pathway proteins and reduced GLUT4 pathway proteins. Rat neonatal cardiomyocytes showed similar results. In conclusion, HDL reversed palmatic acid‐induced lipotoxicity and energy metabolism imbalance in H9c2 cardiomyoblast cells and in neonatal rat cardiomyocyte cells. In our previous study palmitic acid (PA) induced lipotoxicity and switches energy metabolism from CD36 to GLUT4 in H9c2 cells. Low level of high density lipoprotein (HDL) is an independent risk factor for cardiac hypertrophy. Therefore, we in the present study investigated whether HDL can reverse PA induced lipotoxicity in H9c2 cardiomyoblast cells. In conclusion, HDL reversed palmatic acid‐induced lipotoxicity and energy metabolism imbalance in H9c2 cardiomyoblast cells and in neonatal rat cardiomyocyte cells.

  June 16, 2017   doi: 10.1002/jcp.26007   open full text
• Microtubule actin crosslinking factor 1 promotes osteoblast differentiation by promoting β‐catenin/TCF1/Runx2 signaling axis.
  Lifang Hu, Peihong Su, Chong Yin, Yan Zhang, Runzhi Li, Kun Yan, Zhihao Chen, Dijie Li, Ge Zhang, Liping Wang, Zhiping Miao, Airong Qian, Cory J. Xian.
  Journal of Cellular Physiology. June 16, 2017
  Osteoblast differentiation is a multistep process delicately regulated by many factors, including cytoskeletal dynamics and signaling pathways. Microtubule actin crosslinking factor 1 (MACF1), a key cytoskeletal linker, has been shown to play key roles in signal transduction and in diverse cellular processes; however, its role in regulating osteoblast differentiation is still needed to be elucidated. To further uncover the functions and mechanisms of action of MACF1 in osteoblast differentiation, we examined effects of MACF1 knockdown (MACF1‐KD) in MC3T3‐E1 osteoblastic cells on their osteoblast differentiation and associated molecular mechanisms. The results showed that knockdown of MACF1 significantly suppressed mineralization of MC3T3‐E1 cells, down‐regulated the expression of key osteogenic genes alkaline phosphatase (ALP), runt‐related transcription factor 2 (Runx2) and type I collagen α1 (Col Iα1). Knockdown of MACF1 dramatically reduced the nuclear translocation of β‐catenin, decreased the transcriptional activation of T cell factor 1 (TCF1), and down‐regulated the expression of TCF1, lymphoid enhancer‐binding factor 1 (LEF1), and Runx2, a target gene of β‐catenin/TCF1. In addition, MACF1 knockdown increased the active level of glycogen synthase kinase‐3β (GSK‐3β), which is a key regulator for β‐catenin signal transduction. Moreover, the reduction of nuclear β‐catenin amount and decreased expression of TCF1 and Runx2 were significantly reversed in MACF1‐KD cells when treated with lithium chloride, an agonist for β‐catenin by inhibiting GSK‐3β activity. Taken together, these findings suggest that knockdown of MACF1 in osteoblastic cells inhibits osteoblast differentiation through suppressing the β‐catenin/TCF1‐Runx2 axis. Thus, a novel role of MACF1 in and a new mechanistic insight of osteoblast differentiation are uncovered. This article is protected by copyright. All rights reserved

  June 16, 2017   doi: 10.1002/jcp.26059   open full text
• Histomorphology and innate immunity during the progression of osteoarthritis: Does synovitis affect cartilage degradation?
  Huan Wang, Qingguo Wang, Meijuan Yang, Lili Yang, Weili Wang, Haobin Ding, Dong Zhang, Jing Xu, Xuezhang Tang, Haitao Ding, Qingfu Wang.
  Journal of Cellular Physiology. June 15, 2017
  Osteoarthritis (OA) is a common chronic degenerative disease that affects all joints. At present, the pathological processes and mechanisms of OA are still unclear. Innate immunity, a key player in damage to the structure of the joint and the mechanism by which the host attempts to repair OA, affects all pathological stages of the disease. In the present study, our aim was to assess changes in innate immunity during the pathological processes of OA in articular cartilage (AC) and the synovial membrane (SM), which are the major structures in joints, and to systematically examine the histological changes in AC and SM in mild, moderate and severe cases of OA, in order to further speculate about the manner in which the interactions of AC and SM are facilitated by innate immunity. Histological methods (including HE and Safranin O‐fast green staining), immunofluorescent double staining, TUNEL stain, and Western blots were used to assess the morphological changes within AC and SM tissues in healthy and mild, moderate, or severe OA rats. Our results showed that the damage to AC and SM within the joints progressively worsened in different degrees during the course of the disease, and that the innate immune system was closely involved in the AC and SM during each stage of OA. These findings also confirmed that SM may affect the pathological changes in AC through the innate immune system, and therefore affect the progress of OA. In the present study, our aim was to assess changes in innate immunity during the pathological processes of OA in articular cartilage (AC) and the synovial membrane (SM), which are the major structures in joints, and to systematically examine the histological changes in AC and SM in mild, moderate, and severe cases of OA, in order to further speculate about the manner in which the interactions of AC and SM are facilitated by innate immunity.

  June 15, 2017   doi: 10.1002/jcp.26011   open full text
• MicroRNA: A Novel Target of Curcumin in Cancer Therapy.
  Hamed Mirzaei, Aria Masoudifar, Amirhossein Sahebkar, Naser Zare, Javid Sadri Nahand, Bahman Rashidi, Emadodin Mehrabian, Mohsen Mohammadi, Hamid Reza Mirzaei, Mahmoud Reza Jaafari.
  Journal of Cellular Physiology. June 15, 2017
  Curcumin is known as a natural dietary polyphenol which is extracted from Curcuma longa L. It has been shown that curcumin has a variety of pharmacological effects such as antioxidant, anti‐cancer, anti‐inflammatory, and anti‐microbial activities. Anti‐cancer effects of curcumin are due to targeting of a wide range of cellular and molecular pathways involved in cancer pathogenesis including NF‐kB, MAPK, PTEN, P53, and microRNAs (miRNA) network. Multiple lines of evidence have indicated that curcumin exerts its therapeutic effects via regulating miRNA expression (e.g. miR‐1, miR‐7, miR‐9, miR‐34a, miR‐181, miR‐21 and miR‐19) which could lead to the regulation of underlying cellular and molecular pathways involved in cancer pathogenesis. Exosomes are one of the important classes of biological vehicles which could be released from various types of cells such as cancer cells and stem cells and could change the behavior of recipient cells. It has been shown that treatment of cancer cells with different dose of curcumin leads to the release of exosomes containing curcumin. These exosomes could induce anti‐cancer properties in recipient cells and reduce tumor growth. Hence, exosomes containing curcumin could be applied as powerful tools for cancer treatment. Here, we highlighted various miRNAs which could be affected by curcumin in various types of cancer. Moreover, we highlight exosomes containing curcumin as suitable therapeutic tools in cancer therapy. This article is protected by copyright. All rights reserved

  June 15, 2017   doi: 10.1002/jcp.26055   open full text
• Humanized mice: A brief overview on their diverse applications in biomedical research.
  Shigeyoshi Fujiwara.
  Journal of Cellular Physiology. June 15, 2017
  Model animals naturally differ from humans in various respects and results from the former are not directly translatable to the latter. One approach to address this issue is humanized mice that are defined as mice engrafted with functional human cells or tissues. In humanized mice, we can investigate the development and function of human cells or tissues (including their products encoded by human genes) in the in vivo context of a small animal. As such, humanized mouse models have played important roles that cannot be substituted by other animal models in various areas of biomedical research. Although there are obvious limitations in humanized mice and we may need some caution in interpreting the results obtained from them, it is reasonably expected that they will be utilized in increasingly diverse areas of biomedical research, as the technology for preparing humanized mice are rapidly improved. In this review, I will describe the methodology for generating humanized mice and overview their recent applications in various disciplines including immunology, infectious diseases, drug metabolism, and neuroscience. Humanized mice, defined as mice engrafted with functional human cells or tissues, provide an excellent small animal model in which development and function of human cells/tissues can be directly analyzed in vivo. Here I describe the methodology of generating humanized mice and their recent applications in diverse areas of biomedical research.

  June 15, 2017   doi: 10.1002/jcp.26022   open full text
• Estrogen receptor β regulates the tumoral suppressor PTEN to modulate pituitary cell growth.
  Pablo A. Perez, Juan P. Petiti, Florencia Picech, Carolina B. Guido, Liliana dV Sosa, Ezequiel Grondona, Jorge H. Mukdsi, Ana L. De Paul, Alicia I. Torres, Silvina Gutierrez.
  Journal of Cellular Physiology. June 15, 2017
  In this study, we focused on ERβ regulation in the adenohypophysis under different estrogenic milieu, by analyzing whether ER modulates the phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expression and its subcellular localization on anterior pituitary glands from Wistar rats and GH3 lactosomatotroph cells that over‐expressed ERβ. ERβ was regulated in a cyclic manner, and underwent dynamic changes throughout the estrous cycle, with decreased ERβ+ cells in estrus and under E2 treatment, but increased in ovariectomized rats. In addition, the ERα/β ratio increased in estrus and under E2 stimulation, but decreased in ovariectomized rats. Double immunofluorescence revealed that lactotroph and somatotroph ERβ+ were significantly decreased in estrus. Also, variations in the PTEN expression was observed, which was diminished with high E2 conditions but augmented with low E2 milieu. The subcellular localization of this phosphatase was cell cycle‐dependent, with remarkable changes in the immunostaining pattern: nuclear in arrested pituitary cells but cytoplasmic in stimulated cells, and responding differently to ER agonists, with only DPN being able to increase PTEN expression and retaining it in the nucleus. Finally, ERβ over‐expression increased PTEN with a noticeable subcellular redistribution, and with a significant nuclear signal increase in correlation with an increase of cells in G0/G1 phase. These results showed that E2 is able to inhibit ERβ expression and suggests that the tumoral suppressor PTEN might be one of the signaling proteins by which E2, through ERβ, acts to modulate pituitary cell proliferation, thereby adapting endocrine populations in relation with hormonal necessities. Pituitary ERβ and PTEN were regulated in a cyclic manner throughout the rat estrous cycle, diminished with high E2 conditions but augmented with low E2 milieu. The ERβ agonist, DPN, was able to increase pituitary PTEN expression and retaining it in the nucleus. ERβ over‐expression increased PTEN in correlation with an increment of GH3 cells in G0/G1 phase.

  June 15, 2017   doi: 10.1002/jcp.26025   open full text
• Cytoskeletal remodeling and regulation of cell fate in the hypertensive neonatal pulmonary artery in response to stress.
  Dina Johar.
  Journal of Cellular Physiology. June 15, 2017
  Neonatal pulmonary hypertension (PHN) is a lethal progressive disease that occurs in prenatal circulatory transition. Mechanical wall strain caused by cardiac pulsation integrates with hypoxia to generate rapidly progressive myocyte cytoskeleton disassembly and failure to exert force generation. The physiological responses to such an interaction have not been investigated. The persistent phenotype does not respond to traditional vasodilator therapy; hence, there is a need for new treatment strategies to improve the morbidity and mortality outcomes. We reviewed the current research methods, models, and markers of persistent PHN relevant to oxidative and nitrosative stress as well as cell fate commitment, with an emphasis on apoptosis and proliferation. We surveyed potential investigations into the role of senescence in neonatal PHN cell fate decision programming during vasodilator treatment and suggested putative drug targets to improve clinical outcomes. We identified important signaling intermediates of senescence and cell cycle entry regulation in hypertensive pulmonary arterial tissues. Identifying the concerted interplay between ROS/RNS generation, senescence, and inflammatory signaling will lead to therapeutic targets to combat vascular remodeling in PHN; the factor most limiting to treatment responses is irreversible fibrotic thickening. Prevention of the latter will help reduce mortality and morbidity in infants with PHN and prolong the window of opportunity for vasodilator therapies.

  June 15, 2017   doi: 10.1002/jcp.25950   open full text
• Mild mitochondrial uncoupling induces HSL/ATGL‐independent lipolysis relying on a form of autophagy in 3T3‐L1 adipocytes.
  Stéphane Demine, Silvia Tejerina, Benoît Bihin, Marc Thiry, Nagabushana Reddy, Patricia Renard, Martine Raes, Michel Jadot, Thierry Arnould.
  Journal of Cellular Physiology. June 15, 2017
  Obesity is characterized by an excessive triacylglycerol accumulation in white adipocytes. Various mechanisms allowing the tight regulation of triacylglycerol storage and mobilization by lipid droplet‐associated proteins as well as lipolytic enzymes have been identified. Increasing energy expenditure by inducing a mild uncoupling of mitochondria in adipocytes might represent a putative interesting anti‐obesity strategy as it reduces the adipose tissue triacylglycerol content (limiting alterations caused by cell hypertrophy) by stimulating lipolysis through yet unknown mechanisms, limiting the adverse effects of adipocyte hypertrophy. Herein, the molecular mechanisms involved in lipolysis induced by a mild uncoupling of mitochondria in white 3T3‐L1 adipocytes were characterized. Mitochondrial uncoupling‐induced lipolysis was found to be independent from canonical pathways that involve lipolytic enzymes such as HSL and ATGL. Finally, enhanced lipolysis in response to mitochondrial uncoupling relies on a form of autophagy as lipid droplets are captured by endolysosomal vesicles. This new mechanism of triacylglycerol breakdown in adipocytes exposed to mild uncoupling provides new insights on the biology of adipocytes dealing with mitochondria forced to dissipate energy. The enhanced lipolysis observed in 3T3‐L1 adipocytes in response to mitochondrial uncoupling relies on a form of autophagy (and not HSL/ATGL) as lipid droplets are captured by endolysosomal vesicles. This is a new mechanism of triacylglycerol breakdown in adipocytes exposed to mild uncoupling that provides new insights on the biology of adipocytes dealing with mitochondria forced to dissipate energy.

  June 15, 2017   doi: 10.1002/jcp.25994   open full text
• Selective expression of long non‐coding RNAs in a breast cancer cell progression model.
  Kirsten M. Tracy, Coralee E. Tye, Natalie A. Page, Andrew J. Fritz, Janet L. Stein, Jane B. Lian, Gary S. Stein.
  Journal of Cellular Physiology. June 15, 2017
  Long non‐coding RNAs (lncRNAs) are acknowledged as regulators of cancer biology and pathology. Our goal was to perform a stringent profiling of breast cancer cell lines that represent disease progression. We used the MCF‐10 series, which includes the normal‐like MCF‐10A, HRAS‐transformed MCF‐10AT1 (pre‐malignant), and MCF‐10CA1a (malignant) cells, to perform transcriptome wide sequencing. From these data, we have identified 346 lncRNAs with dysregulated expression across the progression series. By comparing lncRNAs from these datasets to those from an additional set of cell lines that represent different disease stages and subtypes, MCF‐7 (early stage, luminal), and MDA‐MB‐231 (late stage, basal), 61 lncRNAs that are associated with breast cancer progression were identified. Querying breast cancer patient data from The Cancer Genome Atlas, we selected a lncRNA, IGF‐like family member 2 antisense RNA 1 (IGFL2‐AS1), of potential clinical relevance for functional characterization. Among the 61 lncRNAs, IGFL2‐AS1 was the most significantly decreased. Our results indicate that this lncRNA plays a role in downregulating its nearest neighbor, IGFL1, and affects migration of breast cancer cells. Furthermore, the lncRNAs we identified provide a valuable resource to mechanistically and clinically understand the contribution of lncRNAs in breast cancer progression. The gene expression patterns of long non‐coding RNAs were profiled in a model of breast cancer progression. The most downregulated long non‐coding RNA, IGFL2‐AS1 was found to affect the expression of neighboring genes and migration of breast cancer cells.

  June 15, 2017   doi: 10.1002/jcp.25997   open full text
• Nociceptor plasticity: A closer look.
  Maria Caterina Pace, Maria Beatrice Passavanti, Lorenzo De Nardis, Fabio Bosco, Pasquale Sansone, Vincenzo Pota, Manlio Barbarisi, Antonio Palagiano, Fabio Arturo Iannotti, Elisabetta Panza, Caterina Aurilio.
  Journal of Cellular Physiology. June 15, 2017
  Nociceptors are receptors specifically involved in detecting a tissue damage and transducing it in an electrical signal. Nociceptor activation provoked by any kind of acute lesion is related to the release of several mediators of inflammation, within the framework of a process defined as “peripheral sensitization.” This results in an exaggerated response to the painful stimulus, clinically defined as “primary hyperalgesia.” The concept of “neuroplasticity” may explain the adaptive mechanisms carried out by the Nervous System in relation to a “harmful” damage; also, neuroplasticity mechanisms are also fundamental for rehabilitative intervention protocols. Here we review several studies that addressed the role of different receptors and ionic channels discovered on nociceptor surface and their role in pain perception. The changes in expression, distribution, and functioning of receptors and ionic channels are thought to be a part of the neuroplasticity property, through which the Nervous System constantly adapts to external stimuli. Moreover, some of the reviewed mediators are also been associated to “central sensitization,” a process that results in pain chronicization when the painful stimulation is particularly prolonged or intense, and lastly leads to the memorization of the uncomfortable painful perception. Peripheral sensitization results in an exaggerated response to the painful stimulus, clinically defined as “primary hyperalgesia.” The concept of “neuroplasticity” may explain the adaptive mechanisms carried out by the Nervous System in relation to a “harmful” damage. Also, Central sensitization, a process that results in pain chronicization when the painful stimulation is particularly prolonged or intense, may as well be intended as a plastic property of the Nervous System.

  June 15, 2017   doi: 10.1002/jcp.25993   open full text
• The effect of substrate stiffness on cancer cell volume homeostasis.
  Meng Wang, Na Chai, Baoyong Sha, Manyun Guo, Jian Zhuang, Feng Xu, Fei Li.
  Journal of Cellular Physiology. June 14, 2017
  Existing studies on the mechanism of cell volume regulation are mainly relevant to ion channels and osmosis in extracellular fluid. Recently, accumulating evidence has shown that cellular mechanical microenvironment also influences the cell volume. Herein, we investigated the regulation of substrate stiffness on the cell volume homeostasis of MCF‐7 cells and their following migration behaviors. We found that cell volume increases with increasing substrate stiffness, which could be affected by blocking the cell membrane anion permeability and dopamine receptor. In addition, the cell migration is significantly inhibited by decreasing the cell volume using tamoxifen and such inhibition effect on migration is enhanced by increasing substrate stiffness. The cell membrane anion permeability might be the linker between cellular mechanical microenvironment and cellular volume homeostasis regulation. This work revealed the regulation of substrate stiffness on cell volume homeostasis for the first time, which would provide a new perspective into the understanding of cancer metastasis and a promising anti‐cancer therapy through regulation of cell volume homeostasis. This work revealed the regulation of substrate stiffness on cell volume homeostasis for the first time, which could be affected by blocking the cell membrane anion permeability and dopamine receptor. In addition, the cell migration is significantly inhibited by decreasing the cell volume using tamoxifen and such inhibition effect on migration is enhanced by increasing substrate stiffness.

  June 14, 2017   doi: 10.1002/jcp.26026   open full text
• Low shear stress induces endothelial reactive oxygen species via the AT1R/eNOS/NO pathway.
  Yuelin Chao, Peng Ye, Linlin Zhu, Xiangquan Kong, Xinliang Qu, Junxia Zhang, Jie Luo, Hongfeng Yang, Shaoliang Chen.
  Journal of Cellular Physiology. June 14, 2017
  Reactive oxygen species (ROS) contribute to many aspects of physiological and pathological cardiovascular processes. However, the underlying mechanism of ROS induction by low shear stress (LSS) remains unclear. Accumulating evidence has shown that the angiotensin II type 1 receptor (AT1R) is involved in inflammation, apoptosis, and ROS production. Our aim was to explore the role of AT1R in LSS‐mediated ROS induction. We exposed human umbilical vein endothelial cells (HUVECs) to LSS (3 dyn/cm2) for different periods of time. Western blotting and immunofluorescence showed that LSS significantly induced AT1R expression in a time‐dependent manner. Using immunohistochemistry, we also noted a similar increase in AT1R expression in the inner curvature of the aortic arch compared to the descending aorta in C57BL/6 mice. Additionally, HUVECs were cultured with a fluorescent probe, either DCFH, DHE or DAF, after being subjected to LSS. Cell chemiluminescence and flow cytometry results revealed that LSS stimulated ROS levels and suppressed nitric oxide (NO) generation in a time‐dependent manner, which was reversed by the AT1R antagonist Losartan. We also found that Losartan markedly increased endothelial NO synthase (eNOS) phosphorylation at Ser(633,1177) and dephosphorylation at Thr(495), which involved AKT and ERK. Moreover, the ROS level was significantly reduced by endogenous and exogenous NO donors (L‐arginine, SNP) and increased by the eNOS inhibitor L‐NAME. Overall, we conclude that LSS induces ROS via AT1R/eNOS/NO. LSS induced ROS via AT1R/eNOS/NO pathway. Low shear stress increased reactive oxygen species via upregulation of AT1R, which was closely related to eNOS phosphorylation and NO level. AKT and ERK was involved in eNOS phosphorylation.

  June 14, 2017   doi: 10.1002/jcp.26016   open full text
• Hypomorphic conditional deletion of E11/Podoplanin reveals a role in osteocyte dendrite elongation.
  Katherine A. Staines, Behzad Javaheri, Peter Hohenstein, Robert Fleming, Ekele Ikpegbu, Erin Unger, Mark Hopkinson, David J. Buttle, Andrew A. Pitsillides, Colin Farquharson.
  Journal of Cellular Physiology. June 14, 2017
  The transmembrane glycoprotein E11/Podoplanin (Pdpn) has been implicated in the initial stages of osteocyte differentiation. However, its precise function and regulatory mechanisms are still unknown. Due to the known embryonic lethality induced by global Pdpn deletion, we have herein explored the effect of bone‐specific Pdpn knockdown on osteocyte form and function in the post‐natal mouse. Extensive skeletal phenotyping of male and female 6‐week‐old Oc‐cre;Pdpnflox/flox (cKO) mice and their Pdpnflox/flox controls (fl/fl) has revealed that Pdpn deletion significantly compromises tibial cortical bone microarchitecture in both sexes, albeit to different extents (p < 0.05). Consistent with this, we observed an increase in stiffness in female cKO mice in comparison to fl/fl mice (p < 0.01). Moreover, analysis of the osteocyte phenotype by phalloidin staining revealed a significant decrease in the dendrite volume (p < 0.001) and length (p < 0.001) in cKO mice in which deletion of Pdpn also modifies the bone anabolic loading response (p < 0.05) in comparison to age‐matched fl/fl mice. Together, these data confirm a regulatory role for Pdpn in osteocyte dendrite formation and as such, in the control of osteocyte function. As the osteocyte dendritic network is known to play vital roles in regulating bone modeling/remodeling, this highlights an essential role for Pdpn in bone homeostasis. E11/Pdpn has been implicated in the initial stages of osteocyte differentiation; however, its precise function and regulatory mechanisms are still unknown. Using a bone‐specific Pdpn knockout mouse, we have confirmed a regulatory role for Pdpn in osteocyte dendrite formation and as such, in the control of osteocyte function.

  June 14, 2017   doi: 10.1002/jcp.25999   open full text
• E2F is involved in radioresistance of carbon ion induced apoptosis via Bax/caspase 3 signal pathway in human hepatoma cell.
  Yi Xie, Jing Si, Yu‐Pei Wang, Hong‐Yan Li, Cui‐Xia Di, Jun‐Fang Yan, Yan‐Cheng Ye, Yan‐Shan Zhang, Hong Zhang.
  Journal of Cellular Physiology. June 13, 2017
  Deletion of p53, most common genetic alteration, is observed in human tumors and reported to lead to improve in cell radioresistance. Heavy‐ion irradiation (IR) could induce p53−/− cancer cells apoptosis. However, little is known regarding the molecular mechanism in this type of cell apoptosis. The present studies have focused on mechanisms state of signaling pathways as an activator of the cell fate decisions induced by heavy ion IR without p53. Carbon ion IR could induce up‐regulation of E2F1 expression in cancer cells. This phenomenon was not observed in X‐ray IR group. Up‐regulation of E2F1 could cause a higher reduction in clonogenic survival, low level of cellular activity, G2/M phase arrest, promotion of apoptosis rate, up‐regulation of phosphor‐Rb, Bax, and cleaved‐caspase 3 proteins expressions without p53. Changes of E2F1 expressions could partly alter radioresistance in cancer cells. The results were suggested that heavy ion IR could induce p53−/− cancer cells apoptosis via E2F1 signal pathway. Our study provides a scientific rationale for the clinical use of heavy ion as radiotherapy in patients with p53‐deficient tumors, which are often resistant to radiotherapy. The results were suggested that heavy ion IR could induce p53−/− cancer cells apoptosis via E2F1 signal pathway. Our study provides a scientific rationale for the clinical use of heavy ion as radiotherapy in patients with p53‐deficient tumors, which are often resistant to radiotherapy.

  June 13, 2017   doi: 10.1002/jcp.26005   open full text
• Combination of nanotechnology with vascular targeting agents for effective cancer therapy.
  Rana Jahanban‐Esfahlan, Khaled Seidi, Behnaz Banimohamad‐Shotorbani, Ali Jahanban‐Esfahlan, Bahman Yousefi.
  Journal of Cellular Physiology. June 13, 2017
  As a young science, nanotechnology promptly integrated into the current oncology practice. Accordingly, various nanostructure particles were developed to reduce drug toxicity and allow the targeted delivery of various diagnostic and therapeutic compounds to the cancer cells. New sophisticated nanosystems constantly emerge to improve the performance of current anticancer modalities. Targeting tumor vasculature is an attractive strategy to fight cancer. Though the idea was swiftly furthered from basic science to the clinic, targeting tumor vasculature had a limited potential in patients, where tumors relapse due to the development of multiple drug resistance and metastasis. The aim of this review is to discuss the advantages of nanosystem incorporation with various vascular targeting agents, including endogen anti‐angiogenic agents; (ii) inhibitors of angiogenesis‐related growth factors, (iii) inhibitors of tyrosine kinase receptors; (iv) inhibitors of angiogenesis‐related signaling pathways, (v) inhibitors of tumor endothelial cell‐associated markers, and (vi) tumor vascular disrupting agents. We also review the efficacy of nanostructures as natural vascular targeting agents. The efficacy of each approach in cancer therapy is further discussed. This article is protected by copyright. All rights reserved

  June 13, 2017   doi: 10.1002/jcp.26051   open full text
• Angiogenesis biomarkers and their targeting ligands as potential targets for tumor angiogenesis.
  Mohammad Mashreghi, Hassan Azarpara, Mahere Rezazade Bazaz, Arash Jafari, Aria Masoudifar, Hamed Mirzaei, Mahmoud Reza Jaafari.
  Journal of Cellular Physiology. June 13, 2017
  Angiogenesis is known as one of the hallmarks in cancer which could play a key role in providing oxygen and nutrients for tumor cells. It has been shown that tumor cannot grow without sufficient development of new blood vessels. Accordingly, targeting angiogenesis, especially endothelial cells, could be considered as a common therapeutic target in tumors and more investigation on already existing biomarkers and potentially new biomarkers of endothelial cells seems to be necessary in cancer therapy. Moreover, the use of effective targeting approaches such as proteins and peptides, aptamers, and small molecules is an important step for targeting biomarkers associated with endothelial cells and angiogenesis in cancer therapy. These agents are FDA approved, or are currently under investigation in pre‐clinical and clinical studies. Among various biomarkers for angiogenesis microRNAs are suitable candidates for target therapy. These molecules play key roles in tumor angiogenesis which exert their effect via targeting a variety of cellular and molecular pathways involved in tumor angiogenesis. Here, we summarize a variety of biomarkers which their expressions or their functions could change the function of endothelial cells in tumor microenvironments. Moreover, we highlighted various therapeutic agents which could target these biomarkers. This article is protected by copyright. All rights reserved

  June 13, 2017   doi: 10.1002/jcp.26049   open full text
• PP2A deactivation is a common event in oral cancer and reactivation by FTY720 shows promising therapeutic potential.
  Bharath K. Velmurugan, Chien‐Hung Lee, Shang‐Lun Chiang, Chun‐Hung Hua, Mei‐Chung Chen, Shu‐Hui Lin, Kun‐Tu Yeh, Ying‐Chin Ko.
  Journal of Cellular Physiology. June 12, 2017
  Protein phosphatase 2A (PP2A) is a tumor suppressor gene, that has been frequently deactivated in many types of cancer. However, its molecular and clinical relevance in oral squamous cell carcinoma (OSCC) remain unclear. Here we show that, PP2A deactivation is a common event in oral cancer cells and hyperphosphorylation in its tyrosine‐307 (Y307) residue contributes to PP2A deactivation. PP2A restoration by FTY720 treatment reduced cell growth and decreased GSK‐3β phosphorylation without significantly altering other PP2A targets. We further detected PP2A phosphorylation in 262 OSCC tissues. Increased expression of p‐PP2A in the tumor tissues was significantly correlated with higher N2/N3‐stage (aOR = 2.1, 95%CI: 1.2–3.8). Patients with high p‐PP2A expression had lower overall survival rates than those with low expression. Hazard ratio analysis showed that, high p‐PP2A expression was significantly associated with mortality density (aOR = 2.2, 95%CI: 1.2–4.0) and lower 10‐year overall survival (p = 0.027) in lymph node metastasis. However, no interaction was observed between p‐PP2A expression and lymph node metastasis. All our results suggest that PP2A is frequently deactivated in oral cancer and determines poor outcome, restoring its expression by FTY720 can be an alternative therapeutic approach in OSCC.

  June 12, 2017   doi: 10.1002/jcp.26001   open full text
• Antitumoral potential, antioxidant activity and carotenoid content of two Southern Italy tomato cultivars extracts: San Marzano and Corbarino.
  Daniela Barone, Letizia Cito, Giuseppina Tommonaro, Agnese A. Abate, Danila Penon, Rocco De Prisco, Antonella Penon, Iris M. Forte, Elisabetta Benedetti, Annamaria Cimini, Paola Indovina, Barbara Nicolaus, Francesca Pentimalli, Antonio Giordano.
  Journal of Cellular Physiology. June 12, 2017
  Gastric cancer represents a diffuse and aggressive neoplasm, whose mortality index is among the highest in the world. Predisposing factors are E‐cadherin mutations, Helicobacter pylori infection, and a diet rich in salted and smoked food, with a low intake of fresh fruits and vegetables. Here, we analyzed the effect of total lipophilic extracts of two Southern Italy tomato varieties, San Marzano and Corbarino, on an in vitro model of gastric cancer, YCC‐1, YCC‐2 and YCC‐3 cell lines, characterized by different aggressiveness. Our results showed a possible role of these two varieties of tomatoes against typical neoplastic features. The treatment with tomato extracts affected cancer cell ability to grow both in adherence and in semisolid medium, reducing also cell migration ability. No toxic effects were observed on non‐tumoral cells. We found, on gastric cancer cell lines, effects on both cell cycle progression and apoptosis modulation. The extent of antineoplastic effects, however, did not seem to correlate with the carotenoid content and antioxidant activity of the two tomato varieties. Our data indicate that San Marzano and Corbarino intake might be further considered as nutritional support not only in cancer prevention, but also for cancer patient diet. We analyzed the effect of total lipophilic extracts of two Southern Italy tomato varieties, San Marzano and Corbarino, on an in vitro model of gastric cancer. The treatment with tomato extracts affected cancer cell ability to grow both in adherence and in semisolid medium, reducing also cell migration ability; these effects were caused by both cell cycle progression and apoptosis modulation. Data showed a possible role of these two varieties of tomatoes against typical neoplastic features.

  June 12, 2017   doi: 10.1002/jcp.25995   open full text
• Involvement of the Warburg effect in non‐tumor diseases processes.
  Zhe Chen, Meiqing Liu, Lanfang Li, Linxi Chen.
  Journal of Cellular Physiology. June 12, 2017
  Warburg effect, as an energy shift from mitochondrial oxidative phosphorylation to aerobic glycolysis, is extensively found in various cancers. Interestingly, increasing researchers show that Warburg effect plays a crucial role in non‐tumor diseases. For instance, inhibition of Warburg effect can alleviate pulmonary vascular remodeling in the process of pulmonary hypertension (PH). Interference of Warburg effect improves mitochondrial function and cardiac function in the process of cardiac hypertrophy and heart failure. Additionally, the Warburg effect induces vascular smooth muscle cell proliferation and contributes to atherosclerosis. Warburg effect may also involve in axonal damage and neuronal death, which are related with multiple sclerosis. Furthermore, Warburg effect significantly promotes cell proliferation and cyst expansion in polycystic kidney disease (PKD). Besides, Warburg effect relieves amyloid β‐mediated cell death in Alzheimer's disease. And Warburg effect also improves the mycobacterium tuberculosis infection. Finally, we also introduce some glycolytic agonists. This review focuses on the newest researches about the role of Warburg effect in non‐tumor diseases, including PH, tuberculosis, idiopathic pulmonary fibrosis (IPF), failing heart, cardiac hypertrophy, atherosclerosis, Alzheimer's diseases, multiple sclerosis, and PKD. Obviously, Warburg effect may be a potential therapeutic target for those non‐tumor diseases. Warburg effect plays a crucial role in non‐tumor diseases. This review focuses on the newest researches about the role of Warburg effect in non‐tumor diseases, including pulmonary hypertension, tuberculosis, idiopathic pulmonary fibrosis, failing heart, cardiac hypertrophy, atherosclerosis, Alzheimer's diseases, multiple sclerosis, and polycystic kidney disease. Obviously, Warburg effect may be a potential therapeutic target for those non‐tumor diseases.

  June 12, 2017   doi: 10.1002/jcp.25998   open full text
• IFN‐gamma priming of adipose‐derived stromal cells at “physiological” hypoxia and under acute hypoxic stress.
  E.R. Andreeva, O.O. Udartseva, O.V. Zhidkova, S.V. Buravkov, M.I. Ezdakova, L.B. Buravkova.
  Journal of Cellular Physiology. June 10, 2017
  Multipotent mesenchymal stromal cells (MSCs) are considered cue regulators of tissue remodelling. Their activity is strongly governed by local milieu, where O2 level is most important. The elevation of inflammatory mediators and acute O2 lowering may additionally modulate MSC activity. In present paper the priming effects of IFN‐gamma on adipose tissue‐derived MSCs (ASCs) at tissue‐related O2 level (5%) and acute hypoxic stress (0.1% O2) were assessed as alterations of ASCs' CFU‐F, proliferation, migration, osteo‐commitment. IFN‐gamma priming provoked ROS elevation, cell growth slowdown, attenuation of both spontaneous and induced osteodifferentiation of tissue O2‐adapted ASCs. The prominent changes in ASC cytoskeleton‐related gene transcription was detected. IFN‐gamma exposure shifted the ASC paracrine profile, suppressing the production of VEGF and IL‐8, while MCP‐1 and IL‐6 were stimulated. Conditioned medium of IFN‐gamma‐primed ASCs did not activate vessel growth in the CAM assay, but induced endothelial cell migration in “wound closure”. Short‐term hypoxia suppressed CFU‐F number, IFN‐gamma‐induced elevation of IL‐6 and endothelial cell migration, while it abolished IFN‐gamma‐provoked VEGF inhibition. After N‐acetyl cysteine treatment ROS level was partly abolished providing additional enhancement of IL‐6 and suppression of IL‐8 and VEGF production. These findings demonstrated that paracrine activity of ASCs in part may be governed by ROS level. Thus, this study first demonstrated that IFN‐gamma priming itself and in combination with acute O2 deprivation could supply dual effects on ASC functions providing both stimulatory and hampering effects. The equilibrium of these factors is a substantial requirement for the execution of MSC remodelling functions. This article is protected by copyright. All rights reserved

  June 10, 2017   doi: 10.1002/jcp.26046   open full text
• The Roles of Signaling Pathways in Bone Repair and Regeneration.
  Maryam Majidinia, Bahman Yousefi.
  Journal of Cellular Physiology. June 07, 2017
  Regenerative medicine has sparked interest in potential strategies for bone repair. Bone defects are widespread and could be caused by trauma, congenital malformations, infections, and surgery. Although bone has a large self‐healing capacity, some defects or fractures are too big to regenerate. To regenerate bone structures which can be used for treatment of patients, bone growth must be induced by a number of bioactive implantable materials, cell types and intracellular and extracellular molecular signaling pathways. Since mesenchymal stem cells (MSCs) and their differentiation during remodeling processes have important roles in bone regeneration, it is believed that understanding molecular signaling pathways involved is crucial to the development of bone implants, bone substitute materials, and cell‐based scaffolds for bone regeneration. In this review, we briefly introduce concepts in fracture repair and regeneration following bone injuries, and then discuss the current clinical methods in bone regeneration. In the next section, we review the involvement of the various key signaling pathways in bone regeneration. This article is protected by copyright. All rights reserved

  June 07, 2017   doi: 10.1002/jcp.26042   open full text
• Curcumin as a potential candidate for treating hyperlipidemia: A review of cellular and metabolic mechanisms.
  Yunes Panahi, Yasin Ahmadi, Manouchehr Teymouri, Thomas P. Johnston, Amirhossein Sahebkar.
  Journal of Cellular Physiology. June 06, 2017
  Curcumin is an herbal polyphenol extensively investigated for antioxidant, anti‐inflammatory, and hypolipidaemic properties. In the present review, the efficacy of curcumin for improving a plasma lipid profile has been evaluated and compared with statins, a well‐known class of medicines for treating hypercholesterolemia and hyperlipidaemia. Curcumin is presumably most effective in reducing triglyceride (TG), while statins are most efficient in lowering low‐density lipoproteins‐cholesterol (LDL‐C). Additionally, various molecular and metabolic mediators of cholesterol and plasma lipid homeostasis are discussed in relation to how they are modulated by curcumin or statins. Overall, curcumin influences the same mediators of plasma lipid alteration as statins do. Almost all the pathways through which cholesterol trafficking takes place are affected by these agents. These include gastrointestinal absorption of dietary cholesterol, hepatocellular removal of plasma cholesterol, the mediators of reverse cholesterol transport, and removal of cholesterol from peripheral tissues. Moreover, the reactive oxygen species (ROS) scavenging potential of curcumin limits the risk of lipid peroxidation that triggers inflammatory responses causing cardiovascular diseases (CVD) and atherosclerosis. Taken together, curcumin could be used as a safe and well‐tolerated adjunct to statins to control hyperlipidaemia more effectively than statins alone. In the present review, the efficacy of curcumin for improving a plasma lipid profile has been evaluated and compared with statins, a well‐known class of medicines for treating hypercholesterolemia and hyperlipidaemia.

  June 06, 2017   doi: 10.1002/jcp.25756   open full text
• TGF‐β inhibits osteogenesis by upregulating the expression of ubiquitin ligase SMURF1 via MAPK‐ERK signaling.
  Xuewu Sun, Ziang Xie, Yan Ma, Xin Pan, Jiying Wang, Zhijun Chen, Peihua Shi.
  Journal of Cellular Physiology. June 05, 2017
  High incidence of osteoporotic fractures emphasizes the necessity of developing effective measures to promote osteogenesis. In our study, we investigated a possible role of MAPK‐ERK signaling in the TGF‐β‐mediated osteoblastic differentiation. Our results indicated that TGF‐β activated the MAPK‐ERK pathway and inhibited osteogenesis in mesenchymal pluripotent cell line, C3H10T1/2, and preosteoblastic cell line, MC3T3 cells. And the downregulation of MAPK‐ERK signaling using pharmacological inhibitor U0126 and RNA interference rescued osteoblast differentiation suppressed by TGF‐β, which was confirmed by Alkaline phosphatase (ALP) staining and alizarrn red staining, and the enhanced expression of osteogenesic markers. Western blotting analysis indicated that TGF‐β induced protein expression of E3 ubiquitin‐protein ligase SMURF1, which contributed to the degradation of RUNX2 and SMAD1 as evidenced by SMURF1 inhibition using RNA interference and proteasome inhibitor MG132. Moreover, we observed that the expression of SMURF1 was decreased, while that of SMAD1 and RUNX2 increased by MAPK‐ERK inhibitor U0126 in TGF‐β‐treated differentiating preosteoblasts, suggesting that MAPK‐ERK regulated the transcription of osteogenesis‐related genes. Furthermore, a synergistic effect between U0126 and bone morphogenic protein (BMP)‐2 on osteoblast differentiation and bone formation was observed both in cell cultures and experimental animals. In conclusion, our results revealed that TGF‐β inhibited osteoblastic differentiation by inducing the MAPK‐ERK pathway which upregulated the expression of ubiquitin ligase SMURF1 and resulted in reduced presence of osteogenic proteins. In addition, the potentiation of BMP‐2 on osteogenic activity by ERK1/2 inhibitor U0126 suggests that it may have potential clinical utility for promoting osteogenesis in bone fracture repair. TGF‐β activates MAPK‐ERK signaling to inhibit osteogenesis through Smurf‐1.MAPK‐ERK inhibitor U0126 and BMP‐2 have a synergistic positive effect on osteoblast differentiation and bone formation.

  June 05, 2017   doi: 10.1002/jcp.25920   open full text
• Swelling‐induced chloride current in glioblastoma proliferation, migration, and invasion.
  Raymond Wong, Wenliang Chen, Xiao Zhong, James T. Rutka, Zhong‐Ping Feng, Hong‐Shuo Sun.
  Journal of Cellular Physiology. June 05, 2017
  Glioblastoma (GBM) remains as the most common and aggressive brain tumor. The survival of GBM has been linked to the aberrant activation of swelling‐induced chloride current ICl,swell. In this study, we investigated the effects of ICl,swell on cell viability, proliferation, and migration in the human GBM cell lines, U251 and U87, using a combination of patch clamp electrophysiology, MTT, colony formation, wound healing assays and Western immunoblotting. First, we showed that the specific inhibitor of ICl,swell, DCPIB, potently reduced the ICl,swell in U87 cells. Next, in both U87 and U251 cells, we found that DCPIB reduced GBM viability, proliferation, colony formation, migration, and invasion. In addition, our Western immunoblot assay showed that DCPIB‐treated U251 cells had a reduction in JAK2, STAT3, and Akt phosphorylation, thus, suggesting that DCPIB potentially suppresses GBM functions through inhibition of the JAK2/STAT3 and PI3K/Akt signaling pathways. Therefore, the ICl,swell may be a potential drug target for GBM. Illustration of the cellular pathways involved in the effects of ICl,swell inhibition on GBM proliferation, migration and invasion.

  June 05, 2017   doi: 10.1002/jcp.25891   open full text
• Bone loss in C57BL/6J‐OlaHsd mice, a substrain of C57BL/6J carrying mutated alpha‐synuclein and multimerin‐1 genes.
  Tamar Liron, Bitya Raphael, Sahar Hiram‐Bab, Itai A. Bab, Yankel Gabet.
  Journal of Cellular Physiology. June 05, 2017
  The inbred mouse strain C57BL/6 is commonly used for the generation of transgenic mouse and is a well established strain in bone research. Different vendors supply different substrains of C57BL/6J as wild‐type animals when genetic drift did not incur any noticeable phenotype. However, we sporadically observed drastic differences in the bone phenotype of “WT” C57BL/6J mice originating from different labs and speculated that these variations are attributable, at least in part, to the variation between C57BL/6J substrains, which is often overlooked. C57BL/6J‐OlaHsd is a commonly used substrain that despite a well defined deletion in the alpha‐synuclein (Snca) and multimerin‐1 (Mmrn1) genes, was reported to display no obvious phenotype and is used as WT control. Here, we compared the bone phenotype of C57BL/6J‐OlaHsd (6J‐OLA) to C57BL/6J‐RccHsd (6J‐RCC) and to the original C57BL/6J (6J‐JAX). Using μCT analysis, we found that 6J‐OLA mice display a significantly lower trabecular bone mass compared to 6J‐RCC and 6J‐JAX. PCR analysis revealed that both the Snca and Mmrn1 genes are expressed in bone tissue of 6J‐RCC animals but not of 6J‐OLA mutants, suggesting either one or both genes play a role in bone metabolism. In vitro analysis demonstrated increase in osteoclasts number and decreased osteoblast mineralization in cells derived from 6J‐OLA compared with 6J‐RCC. Our data may shed light on unexplained differences in basal bone measurements between different research centers and reiterate the importance of specifying the exact substrain type. In addition, our findings describe the physiological role for Mmrn1 and/or Snca in bone remodeling. The C57Bl/6J‐OlaHsd substrain commonly considered as wild‐type mice, display a low bone mass phenotype compared to the original 6J substrain. These differences are attributable to the physiological role of alpha‐synuclein and multimerin in osteoblast and osteoclast functions. These findings also reiterate the critical importance of using littermates as controls for transgenic mice.

  June 05, 2017   doi: 10.1002/jcp.25895   open full text
• Phosphoregulation of K+‐Cl− cotransporters during cell swelling: Novel insights.
  Rachelle Frenette‐Cotton, Andrée‐Anne Marcoux, Alexandre P. Garneau, Micheline Noel, Paul Isenring.
  Journal of Cellular Physiology. June 05, 2017
  The K+‐Cl− cotransporters (KCCs) belong to the cation‐Cl− cotransporter family and consist of four isoforms and many splice variants. Their main role is to promote electroneutral efflux of K+ and Cl− ions across the surface of many cell types and, thereby, to regulate intracellular ion concentration, cell volume, and epithelial salt movement. These transport systems are induced by an increase in cell volume and are less active at lower intracellular [Cl−] (Cli), but the mechanisms at play are still ill‐defined. In this work, we have exploited the Xenopus laevis expression system to study the role of lysine‐deficient protein kinases (WNKs), protein phosphatases 1 (PP1s), and SPS1‐related proline/alanine‐rich kinase (SPAK) in KCC4 regulation during cell swelling. We have found that WNK4 and PP1 regulate KCC4 activity as part of a common signaling module, but that they do not exert their effects through SPAK or carrier dephosphorylation. We have also found that the phosphatases at play include PP1α and PP1γ1, but that WNK4 acts directly on the PP1s instead of the opposite. Unexpectedly, however, both cell swelling and a T926A substitution in the C‐terminus of full‐length KCC4 led to higher levels of heterologous K+‐Cl− cotransport and overall carrier phosphorylation. These results imply that the response to cell swelling must also involve allosteric‐sensitive kinase‐dependent phosphoacceptor sites in KCC4. They are thus partially inconsistent with previous models of KCC regulation. We have found that both cell swelling and a T926A substitution in the C‐terminus of full‐lenght KCC4 lead to higher levels of heterologous K‐Cl cotranport by increasing overall carrier phosphorylation. These results imply that the response to cell swelling could involve allosteric‐sensitive kinase‐dependent phosphoacceptor sites in KCC4. They are thus partially inconsistent with previous models of KCC regulation.

  June 05, 2017   doi: 10.1002/jcp.25899   open full text
• Bone marrow adipocytes support haematopoietic stem cell survival.
  Domenico Mattiucci, Giulia Maurizi, Valerio Izzi, Lorenzo Cenci, Marco Ciarlantini, Stefania Mancini, Emanuela Mensà, Raffaele Pascarella, Marco Vivarelli, Attilio Olivieri, Pietro Leoni, Antonella Poloni.
  Journal of Cellular Physiology. June 02, 2017
  In bone marrow (BM), haematopoietic elements are mingled with adipocytes (BM‐A), which are the most abundant stromal component in the niche. BM‐A progressively increase with ageing, eventually occupying up to 50% of BM cavities. In this work, the role played by BM‐A was explored by studying primary human BM‐A isolated from hip surgery patients at the molecular level, through microarray analysis, and at the functional level, by assessing their relationship with primary human haematopoietic stem cells (HSC) by the long‐term culture initiating cell (LTC‐IC) assay. Findings demonstrated that BM‐A are capable of supporting HSC survival in the LTC‐IC assay, since after 5 weeks of co‐culture, HSC were still able to proliferate and differentiate. Furthermore, critical molecules such as C‐X‐C motif chemokine 12 (CXCL12), interleukin (IL)‐8, colony‐stimulating factor 3 (CSF3), and leukaemia inhibitory factor (LIF), were expressed at similar levels in BM‐A and in primary human BM mesenchymal stromal cells (BM‐MSC), whereas IL‐3 was higher in BM‐A. Interestingly, BM‐A displayed a different gene expression profile compared with subcutaneous adipose tissue adipocytes (AT‐A) collected from abdominal surgery patients, especially in terms of regulation of lipid metabolism, stemness genes and white‐to‐brown differentiation pathways. Accordingly, analysis of the gene pathways involved in haematopoiesis regulation showed that BM‐A are more closely related to BM‐MSC than to AT‐A. The present data suggest that BM‐A play a supporting role in the haematopoietic niche and directly sustain HSC survival. This article is protected by copyright. All rights reserved

  June 02, 2017   doi: 10.1002/jcp.26037   open full text
• Hydroxychloroquine affects bone resorption both in vitro and in vivo.
  Tim Both, M. Carola Zillikens, Marijke Koedam, Marijn Vis, Wai‐Kwan Lam, Angelique E.A. M. Weel, Johannes P.T. M. van Leeuwen, P. Martin van Hagen, Bram C.J. van der Eerden, Paul L.A. van Daele.
  Journal of Cellular Physiology. May 30, 2017
  We recently showed that patients with primary Sjögren syndrome (pSS) have significantly higher bone mineral density (BMD) compared to healthy controls. The majority of those patients (69%) was using hydroxychloroquine (HCQ), which may have favorable effects on BMD. The aim of the study was to evaluate whether HCQ modulates osteoclast function. Osteoclasts were cultured from PBMC‐sorted monocytes for 14 days and treated with different HCQ doses (control, 1 and 5 µg/ml). TRAP staining and resorption assays were performed to evaluate osteoclast differentiation and activity, respectively. Staining with an acidification marker (acridine orange) was performed to evaluate intracellular pH at multiple timepoints. Additionally, a fluorescent cholesterol uptake assay was performed to evaluate cholesterol trafficking. Serum bone resorption marker β‐CTx was evaluated in rheumatoid arthritis patients. HCQ inhibits the formation of multinuclear osteoclasts and leads to decreased bone resorption. Continuous HCQ treatment significantly decreases intracellular pH and significantly enhanced cholesterol uptake in mature osteoclasts along with increased expression of the lowdensity lipoprotein receptor. Serum β‐CTx was significantly decreased after six months of HCQ treatment. In agreement with our clinical data, we demonstrate that HCQ suppresses bone resorption in vitro and decreases the resorption marker β‐CTx in vivo. We also showed that HCQ decreases the intracellular pH in mature osteoclasts and stimulates cholesterol uptake, suggesting that HCQ induces osteoclastic lysosomal membrane permeabilization (LMP) leading to decreased resorption without changes in apoptosis. We hypothesize that skeletal health of patients with increased risk of osteoporosis and fractures may benefit from HCQ by preventing BMD loss. This article is protected by copyright. All rights reserved

  May 30, 2017   doi: 10.1002/jcp.26028   open full text
• Microtubules Regulate Brush Border Formation.
  Facundo M. Tonucci, Anabela Ferretti, Evangelina Almada, Pamela Cribb, Rodrigo Vena, Florencia Hidalgo, Cristián Favre, Matt J. Tyska, Irina Kaverina, M. Cecilia Larocca.
  Journal of Cellular Physiology. May 26, 2017
  Most epithelial cells contain apical membrane structures associated to bundles of actin filaments, which constitute the brush border. Whereas microtubule participation in the maintenance of the brush border identity has been characterized, their contribution to de novo microvilli organization remained elusive. Hereby, using a cell model of individual enterocyte polarization, we found that nocodazole induced microtubule depolymerization prevented the de novo brush border formation. Microtubule participation in brush border actin organization was confirmed in polarized kidney tubule MDCK cells. We also found that centrosome, but not Golgi derived microtubules, were essential for the initial stages of brush border development. During this process, microtubule plus ends acquired an early asymmetric orientation towards the apical membrane, which clearly differs from their predominant basal orientation in mature epithelia. In addition, overexpression of the microtubule plus ends associated protein CLIP170, which regulate actin nucleation in different cell contexts, facilitated brush border formation. In combination, the present results support the participation of centrosomal microtubule plus ends in the activation of the polarized actin organization associated to brush border formation, unveiling a novel mechanism of microtubule regulation of epithelial polarity. This article is protected by copyright. All rights reserved

  May 26, 2017   doi: 10.1002/jcp.26033   open full text
• The nucleus is irreversibly shaped by motion of cell boundaries in cancer and non‐cancer cells.
  Vincent J. Tocco, Yuan Li, Keith G. Christopher, James H. Matthews, Varun Aggarwal, Lauren Paschall, Hendrik Luesch, Jonathan D Licht, Richard B. Dickinson, Tanmay P. Lele.
  Journal of Cellular Physiology. May 25, 2017
  Actomyosin stress fibers impinge on the nucleus and can exert compressive forces on it. These compressive forces have been proposed to elongate nuclei in fibroblasts, and lead to abnormally shaped nuclei in cancer cells. In these models, the elongated or flattened nuclear shape is proposed to store elastic energy. However, we found that deformed shapes of nuclei are unchanged even after removal of the cell with micro‐dissection, both for smooth, elongated nuclei in fibroblasts and abnormally shaped nuclei in breast cancer cells. The lack of shape relaxation implies that the nuclear shape in spread cells does not store any elastic energy, and the cellular stresses that deform the nucleus are dissipative, not static. During cell spreading, the deviation of the nucleus from a convex shape increased in MDA‐MB‐231 cancer cells, but decreased in MCF‐10A cells. Tracking changes of nuclear and cellular shape on micropatterned substrata revealed that fibroblast nuclei deform only during deformations in cell shape and only in the direction of nearby moving cell boundaries. We propose that motion of cell boundaries exert a stress on the nucleus, which allows the nucleus to mimic cell shape. The lack of elastic energy in the nuclear shape suggests that nuclear shape changes in cells occur at constant surface area and volume. This article is protected by copyright. All rights reserved

  May 25, 2017   doi: 10.1002/jcp.26031   open full text
• Chick derived induced pluripotent stem cells by the poly‐cistronic transposon with enhanced transcriptional activity.
  Masafumi Katayama, Takashi Hirayama, Tetsuya Tani, Katsuhiko Nishimori, Manabu Onuma, Tomokazu Fukuda.
  Journal of Cellular Physiology. May 24, 2017
  Induced pluripotent stem (iPS) cell technology lead terminally differentiated cells into the pluripotent stem cells through the expression of defined reprogramming factors. Although, iPS cells have been established in a number of mammalian species, including mouse, human, and monkey, studies on iPS cells in avian species are still very limited. To establish chick iPS cells, six factors were used within the poly‐cistronic reprogramming vector (PB‐R6F), containing M3O (MyoD derived transactivation domain fused with Oct3/4), Sox2, Klf4, c‐Myc, Lin28, and Nanog. The PB‐R6F derived iPS cells were alkaline‐phosphatase and SSEA‐1 positive, which are markers of pluripotency. Elevated levels of endogenous Oct3/4 and Nanog genes were detected in the established iPS cells, suggesting the activation of the FGF signaling pathway is critical for the pluripotent status. Histological analysis of teratoma revealed that the established chick iPS cells have differentiation ability into three‐germ‐layer derived tissues. This is the first report of establishment of avian derived iPS cells with a single poly‐cistronic transposon based expression system. The establishment of avian derived iPS cells could contribute to the genetic conservation and modification of avian species. This is the first report of establishment of avian derived iPS cells with a single poly‐cistronic transposon based expression system. Our established iPS cells depend on the activation of FGF signaling pathway, but does not to the LIF signaling.

  May 24, 2017   doi: 10.1002/jcp.25947   open full text
• DJ‐1/Park7 Sensitive Na+/H+ Exchanger 1 (NHE1) in CD4+ T Cells.
  Yuetao Zhou, Xiaolong Shi, Hong Chen, Shaqiu Zhang, Madhuri S. Salker, Andreas F. Mack, Michael Föller, Tak W. Mak, Yogesh Singh, Florian Lang.
  Journal of Cellular Physiology. May 24, 2017
  DJ‐1/Park7 is a redox‐sensitive chaperone protein counteracting oxidation and presumably contributing to the control of oxidative stress responses and thus inflammation. DJ‐1 gene deletion exacerbates the progression of Parkinson's disease presumably by augmenting oxidative stress. Formation of reactive oxygen species (ROS) is paralleled by activation of the Na+/H+ exchanger 1 (NHE1). ROS formation in CD4+ T cells plays a decisive role in regulating inflammatory responses. In the present study, we explored whether DJ‐1 is expressed in CD4+ T cells, and affects ROS production as well as NHE1 in those cells. To this end, DJ‐1 and NHE1 transcript, and protein levels were quantified by qRT‐PCR and Western blotting, respectively, intracellular pH (pHi) utilizing bis‐(2‐carboxyethyl)‐5‐(and‐6)‐carboxyfluorescein (BCECF) fluorescence, NHE activity from realkalinization after an ammonium pulse, and ROS production utilizing 2′,7′ −dichlorofluorescin diacetate (DCFDA) fluorescence. As a result DJ‐1 was expressed in CD4+ T cells. ROS formation, NHE1 transcript levels, NHE1 protein, and NHE activity were higher in CD4+ T cells from DJ‐1 deficient mice than in CD4+ T cells from wild type mice. Antioxidant N‐acetyl‐cysteine (NAC) and protein tyrosine kinase (PTK) inhibitor staurosporine decreased the NHE activity in DJ‐1 deficient CD4+ T cells, and blunted the difference between DJ‐1−/− and DJ‐1+/+ CD4+ T cells, an observation pointing to a role of ROS in the up‐regulation of NHE1 in DJ‐1−/− CD4+ T cells. In conclusion, DJ‐1 is a powerful regulator of ROS production as well as NHE1 expression and activity in CD4+ T cells. J. Cell. Physiol. 9999: 1–10, 2017. © 2017 Wiley Periodicals, Inc. Our study described that DJ‐1 deficient CD4+ T cells leads to upregulation of NHE1 and enhanced ROS production, therefore results in increase in NHE1 activity. NHE1 regulation in DJ‐1 deficient CD4+ T cells is dependent on ROS and PTK signaling. Thus, our data first time described that link between DJ‐1 proteins with responses on ROS production and NHE1 functions.

  May 24, 2017   doi: 10.1002/jcp.25516   open full text
• Characterization of Runx2 phosphorylation sites required for TGF‐β1‐mediated stimulation of matrix metalloproteinase‐13 expression in osteoblastic cells.
  Balasubramanian Arumugam, Mariappanadar Vairamani, Nicola C. Partridge, Nagarajan Selvamurugan.
  Journal of Cellular Physiology. May 24, 2017
  Transforming growth factor‐beta1 (TGF‐β1), a highly abundant growth factor in skeletal tissues, stimulates matrix metalloproteinase‐13 (MMP‐13) expression in osteoblastic cells. MMP‐13 plays a critical role in bone remodeling. Runx2, a bone transcription factor, is required for TGF‐β1‐mediated stimulation of MMP‐13 expression in osteoblastic cells. In this study, the molecular mechanism responsible for TGF‐β1‐stimulation of MMP‐13 expression via Runx2 in osteoblastic cells was elucidated. TGF‐β1 stimulated the phosphorylation of Runx2 at serine amino acids, and ERK inhibition blocked this effect in rat (UMR106‐01) and human (MG‐63) osteoblastic cells. Pretreatment with okadaic acid, a serine‐threonine phosphatase inhibitor, increased Runx2 serine phosphorylation in osteoblastic cells. When cells were pretreated with an ERK inhibitor, TGF‐β1‐mediated stimulation of MMP‐13 mRNA expression decreased. Nano‐ESI/LC/MS analysis identified that TGF‐β1 stimulates Runx2 phosphorylation at three serine amino acids. Transient transfection of mouse mesenchymal stem cells (C3H10T1/2) with Runx2 serine mutant constructs decreased TGF‐β1‐mediated Runx2 serine phosphorylation. A luciferase reporter assay identified that TGF‐β1 stimulated MMP‐13 promoter activity in these cells only in the presence of the wild Runx2 construct, and not with mutant Runx2. Thus, TGF‐β1 stimulates the phosphorylation of Runx2 at three serine amino acids, and this event is required for MMP‐13 expression in osteoblastic cells. Hence, this study contributes to the knowledge of events governing bone remodeling and bone‐related diseases. A schematic diagram representing the TGF‐β1‐induced stimulation of ERK‐dependent Runx2 phosphorylation, with the corresponding sites, required for MMP‐13 promoter activation in human osteoblast cells.

  May 24, 2017   doi: 10.1002/jcp.25964   open full text
• Isorhapontigenin induced cell growth inhibition and apoptosis by targeting EGFR‐related pathways in prostate cancer.
  Cuicui Zhu, Qingyi Zhu, Zhaomeng Wu, Yingying Yin, Dan Kang, Shan Lu, Ping Liu.
  Journal of Cellular Physiology. May 24, 2017
  Isorhapontigenin (ISO), a naturally phytopolyphenol compound existing in Chinese herb, apples, and various vegetables, has attracted extensive interest in recent years for its diverse pharmacological characteristics. Increasing evidences reveal that ISO can inhibit cancer cell growth by induced apoptosis, however, the molecular mechanisms is not fully understood. In this study, we found for the first time that ISO apparently induced cell growth inhibition and apoptosis by targeting EGFR and its downstream signal pathways in prostate cancer (PCa) cells both in vitro and in vivo, whereas no obviously effect on normal prostate cells. From the results, we found that ISO competitively targeted EGFR with EGF and inhibited EGFR auto‐phosphorylation, and then decreased the levels of p‐Erk1/2, p‐PI3 K, and p‐AKT, and further induced down‐regulation of p‐FOXO1 and promoted FOXO1 nuclear translocation; and finally resulted in a significantly up‐regulation of Bim/p21/27/Bax/cleaved Caspase‐3/cleaved PARP‐1 and a markedly down‐regulation of Sp1/Bcl‐2/XIAP/Cyclin D1. Moreover, our experimental data demonstrated that treatment of ISO decreased protein level of AR via both inhibiting the expression of AR gene and promoting the ubiquitination/degradation of AR proteins in proteasome. In vivo, we also found that ISO inhibited the growth of subcutaneous xenotransplanted tumor in nude mice by inducing PCa cell growth inhibition and apoptosis. Taken together, all findings here clearly implicated that EGFR‐related signal pathways, including EGFR‐PI3K‐Akt and EGFR‐Erk1/2 pathways, were involved in ISO‐induced cell growth inhibition and apoptosis in PCa cells, providing a more solid theoretical basis for the application of ISO to treat patients with prostate cancer in clinic. In this study, we find for the first time that ISO competitively targets EGFR with EGF and inhibits the auto‐phosphorylation of EGFR at Tyr1173, resulting in inactivation of EGFR and its downstream signal pathways, including EGFR/PI3K/Akt pathway, and EGFR/Erk pathway; and finally induce cell growth inhibition and apoptosis by further activating FOXO1 and inactivating androgen receptor (AR) in prostate cancer cells.

  May 24, 2017   doi: 10.1002/jcp.25968   open full text
• Hematopoietic stem/progenitor cell differentiation towards myeloid lineage is modulated by LIGHT/LIGHT receptor signaling.
  Weikai Chen, Xin Lv, Changlong Liu, Ruoping Chen, Jianhe Liu, Haiyan Dai, Gang‐Ming Zou.
  Journal of Cellular Physiology. May 24, 2017
  The cytokine LT‐related inducible ligand that competes for glycoprotein D binding to herpesvirus entry mediator on T cells (LIGHT) is a member of the tumor necrosis factor (TNF) superfamily. It is expressed primarily on activated T lymphocytes, and detectable on monocytes, granulocytes, and immune dendritic cells. It mainly plays a role in immune regulation including T cell activation and dendritic cell maturation. We recently reported its role as an inducer in embryonic stem cell differentiation, but its role in regulation of adult stem cell has not been defined. In the present study, we examined the expression of LIGHT receptor in Lin−c‐kit+Sca‐1+ hematopoietic stem/progenitor cells (HSC/HPCs). We found that HSC express HVEM, a LIGHT receptor, on its surface. We further identified the role of LIGHT in promoting myeloid differentiation of HSCs driven by granulocyte‐monocyte colony stimulating factor (GM‐CSF). Further studies showed that LIGHT enhances both GM‐CSF and GM‐CSF receptor (GM‐CSFR) expression in HSCs. LIGHT stimulation increases PU.1 expression in HSC/HPCs. In vivo administration of LIGHT increases the colony‐forming unit‐granulocyte/monocyte (CFU‐GM) colony formation and plasma GM‐CSF level. Altogether, the data suggest LIGHT promote myeloid differentiation of HSC/HPCs. In the present study, we examined the expression of LIGHT receptor in Lin‐c‐kit+Sca‐1+ hematopoietic stem/progenitor cells (HSC/HPCs). We found that HSC express HVEM, a LIGHT receptor, on its surface. We further identified the role of LIGHT in promoting myeloid differentiation of HSCs driven by granulocyte‐monocyte colony stimulating factor (GM‐CSF).

  May 24, 2017   doi: 10.1002/jcp.25967   open full text
• Acyl‐CoA synthetase short‐chain family member 2 (ACSS2) is regulated by SREBP‐1 and plays a role in fatty acid synthesis in caprine mammary epithelial cells.
  Huifen Xu, Jun Luo, Gongzhen Ma, Xueying Zhang, Dawei Yao, Ming Li, Juan J. Loor.
  Journal of Cellular Physiology. May 24, 2017
  Sterol regulatory element binding protein 1 (SREBP‐1) is well‐known as the master regulator of lipogenesis in rodents. Acyl‐CoA synthetase short‐chain family member 2 (ACSS2) plays a key role in lipogenesis by synthesizing acetyl‐CoA from acetate for lipogenesis. ATP citrate lyase (ACLY) catalyzes the conversion of citrate and coenzyme A to acetyl‐CoA, hence, it is also important for lipogenesis. Although ACSS2 function in cancer cells has been elucidated, its essentiality in ruminant mammary lipogenesis is unknown. Furthermore, ACSS2 gene promoter and its regulatory mechanisms have not known. Expression of ACSS2 was high in lipid synthesizing tissues, and its expression increased during lactation compared with non‐lactating period. Simultaneous knockdown of both ACSS2 and ACLY by siRNA in primary goat mammary epithelial cells decreased (p < 0.05) the mRNA abundance of genes associated with de novo fatty acid synthesis (FASN, ACACA, SCD1) and triacylglycerol (TAG) synthesis (DGAT1, DGAT2, GPAM, and AGPAT6). Genes responsible for lipid droplet formation and secretion (PLIN2 and PLIN3) and fatty acid oxidation (ATGL, HSL, ACOX, and CPT1A) all decreased (p < 0.05) after ACSS2 and ACLY knockdown. Total cellular TAG content and lipid droplet formation also decreased. Use of a luciferase reporter assay revealed a direct regulation of ACSS2 by SREBP‐1. Furthermore, SREBP‐1 interacted with an SRE (SREBP response element) spanning at −475 to −483 bp on the ACSS2 promoter. Taken together, our results revealed a novel pathway that SREBP‐1 may regulate fatty acid and TAG synthesis by regulating the expression of ACSS2. The expression of ACSS2 is crucial for triacylglycerol synthesis in goat mammary epithelial cells. Its expression is controlled by the transcription factor SREBP‐1 through an SRE element on the promoter region.

  May 24, 2017   doi: 10.1002/jcp.25954   open full text
• End stage renal disease‐induced hypercalcemia may promote aortic valve calcification via Annexin VI enrichment of valve interstitial cell derived‐matrix vesicles.
  Lin Cui, Nabil A. Rashdan, Dongxing Zhu, Elspeth M. Milne, Paul Ajuh, Gillian Milne, Miep H. Helfrich, Kelvin Lim, Sai Prasad, Daniel A. Lerman, Alex T. Vesey, Marc R. Dweck, William S. Jenkins, David E. Newby, Colin Farquharson, Vicky E. Macrae.
  Journal of Cellular Physiology. May 24, 2017
  Patients with end‐stage renal disease (ESRD) have elevated circulating calcium (Ca) and phosphate (Pi), and exhibit accelerated progression of calcific aortic valve disease (CAVD). We hypothesized that matrix vesicles (MVs) initiate the calcification process in CAVD. Ca induced rat valve interstitial cells (VICs) calcification at 4.5 mM (16.4‐fold; p < 0.05) whereas Pi treatment alone had no effect. Ca (2.7 mM) and Pi (2.5 mM) synergistically induced calcium deposition (10.8‐fold; p < 0.001) in VICs. Ca treatment increased the mRNA of the osteogenic markers Msx2, Runx2, and Alpl (p < 0.01). MVs were harvested by ultracentrifugation from VICs cultured with control or calcification media (containing 2.7 mM Ca and 2.5 mM Pi) for 16 hr. Proteomics analysis revealed the marked enrichment of exosomal proteins, including CD9, CD63, LAMP‐1, and LAMP‐2 and a concomitant up‐regulation of the Annexin family of calcium‐binding proteins. Of particular note Annexin VI was shown to be enriched in calcifying VIC‐derived MVs (51.9‐fold; p < 0.05). Through bioinformatic analysis using Ingenuity Pathway Analysis (IPA), the up‐regulation of canonical signaling pathways relevant to cardiovascular function were identified in calcifying VIC‐derived MVs, including aldosterone, Rho kinase, and metal binding. Further studies using human calcified valve tissue revealed the co‐localization of Annexin VI with areas of MVs in the extracellular matrix by transmission electron microscopy (TEM). Together these findings highlight a critical role for VIC‐derived MVs in CAVD. Furthermore, we identify calcium as a key driver of aortic valve calcification, which may directly underpin the increased susceptibility of ESRD patients to accelerated development of CAVD. Matrix vesicle (MV) deposition in the extracellular matrix of human calcified aortic valve tissue. Aortic valve calcification was confirmed by Alizarin red and Von Kossa staining. Arrows indicate positive areas of staining. Transmission electron microscopy (TEM) shows MVs with spindle‐like projections resembling hydroxyapatite crystal needles in calcified tissue, and “empty” vesicle structures in control tissue (arrows). Scale bars = 500 µm (Alizarin red/Von Kossa); 500 nm (TEM).

  May 24, 2017   doi: 10.1002/jcp.25935   open full text
• Stimulation of oral fibroblast chemokine receptors identifies CCR3 and CCR4 as potential wound healing targets.
  Jeroen K. Buskermolen, Sanne Roffel, Susan Gibbs.
  Journal of Cellular Physiology. May 23, 2017
  The focus of this study was to determine which chemokine receptors are present on oral fibroblasts and whether these receptors influence proliferation, migration, and/or the release of wound healing mediators. This information may provide insight into the superior wound healing characteristics of the oral mucosa. The gingiva fibroblasts expressed 12 different chemokine receptors (CCR3, CCR4, CCR6, CCR9, CCR10, CXCR1, CXCR2, CXCR4, CXCR5, CXCR7, CX3CR1, and XCR1), as analyzed by flow cytometry. Fourteen corresponding chemokines (CCL5, CCL15, CCL20, CCL22, CCL25, CCL27, CCL28, CXCL1, CXCL8, CXCL11, CXCL12, CXCL13, CX3CL1, and XCL1) were used to study the activation of these receptors on gingiva fibroblasts. Twelve of these fourteen chemokines stimulated gingiva fibroblast migration (all except for CXCL8 and CXCL12). Five of the chemokines stimulated proliferation (CCL5/CCR3, CCL15/CCR3, CCL22/CCR4, CCL28/CCR3/CCR10, and XCL1/XCR1). Furthermore, CCL28/CCR3/CCR10 and CCL22/CCR4 stimulation increased IL‐6 secretion and CCL28/CCR3/CCR10 together with CCL27/CCR10 upregulated HGF secretion. Moreover, TIMP‐1 secretion was reduced by CCL15/CCR3. In conclusion, this in‐vitro study identifies chemokine receptor‐ligand pairs which may be used in future targeted wound healing strategies. In particular, we identified the chemokine receptors CCR3 and CCR4, and the mucosa specific chemokine CCL28, as having an predominant role in oral wound healing by increasing human gingiva fibroblast proliferation, migration, and the secretion of IL‐6 and HGF and reducing the secretion of TIMP‐1. Gingiva fibroblasts express 12 chemokine receptors. Fourteen chemokines were used to study proliferation (top), migration (middle), and secretion of wound healing mediators by gingiva fibroblasts (bottom). Predominantly CCR3 and CCR4 stimulation influenced these oral wound healing parameters.

  May 23, 2017   doi: 10.1002/jcp.25946   open full text
• APN/CD13 is over‐expressed by Psoriatic fibroblasts and is modulated by CGRP and IL‐4 but not by retinoic acid treatment.
  Pascale Gerbaud, Jean Guibourdenche, Rafika Jarray, Marc Conti, Patricia Palmic, Stéphanie Leclerc‐Mercier, Julie Bruneau, Olivier Hermine, Yves Lepelletier, Françoise Raynaud.
  Journal of Cellular Physiology. May 23, 2017
  Psoriasis vulgaris is a common skin inflammatory disease characterized by recurrent flare episodes associated with scaly well‐demarcated skin plaques. Skin biopsies from psoriatic patients with high PASI score (22.67 ± 8.67) and from HD were used to study APN/CD13. APN/CD13 is over‐expressed in LP and nLP compare to HD skins and fibroblasts. This over‐expression is positively correlated with specific enzymatic activity enhancement. However, discrepancies between APN/CD13 expression in LP and nLP prompt us to focus our study on APN/CD13 modulation. Calcitonin Gene Related Peptide (CGRP), a neuropeptide, positively modulated expression and activity of APN/CD13. CGRP consistently induced IL4 secretion, which is also involved in the increase of APN/CD13 expression and activity, which is significantly reversed using IL‐4 blocking antibody. Surprisingly, retinoic acid altered the APN/CD13 enzymatic activity only in nLP fibroblasts without modification of APN/CD13 expression. APN/CD13 is over‐expressed on psoriatic fibroblasts and exerted high level of activity compare to HD fibroblasts. Taken together, several factors such as CGRP and IL‐4 acted on positive regulation of APN/CD13 expression and activity. This study highlighted the interest of APN/CD13 as a new potential target, which should be investigated in psoriasis. APN/CD13 aminopeptidase‐N is upregalated in sporiatic skins compare to healthy donors. APN/CD13 is mostly increased in nLP (non lesional psoriatic) compare to LP (lesional psoriatic) skins. APN/CD13 is positively regulated by IL‐4 and CGRP.

  May 23, 2017   doi: 10.1002/jcp.25941   open full text
• Therapeutic potential of novel formulated forms of curcumin in the treatment of breast cancer by the targeting of cellular and physiological dysregulated pathways.
  Amir Tajbakhsh, Malihe Hasanzadeh, Mehdi Rezaee, Mostafa Khedri, Majid Khazaei, Soodabeh ShahidSales, Gordon A. Ferns, Seyed Mahdi Hassanian, Amir Avan.
  Journal of Cellular Physiology. May 23, 2017
  Breast cancer is among the most important causes of cancer related death in women. There is a need for novel agents for targeting key signaling pathways to either improve the efficacy of the current therapy, or reduce toxicity. There is some evidence that curcumin may have antitumor activity in breast cancer. Several clinical trials have investigated its activity in patients with breast cancer, including a recent trial in breast cancer patients receiving radiotherapy, in whom it was shown that curcumin reduced the severity of radiation dermatitis, although it is associated with low bioavailability. Several approaches have been developed to increase its absorption rate (e.g., nano crystals, liposomes, polymers, and micelles) and co‐delivery of curcumin with adjuvants as well as different conjugation to enhance its bioavailability. In particular, micro‐emulsions is an option for transdermal curcumin delivery, which has been reported to increase its absorption. Lipid‐based nano‐micelles is another approach to enhance curcumin absorption via gastrointestinal tract, while polymer‐based nano‐formulations (e.g., poly D, L‐lactic‐co‐glycolic [PLGA]) allows the release of curcumin at a sustained level. This review summarizes the current data of the therapeutic potential of novel formulations of curcumin with particular emphasis on recent preclinical and clinical studies in the treatment of breast cancer.

  May 23, 2017   doi: 10.1002/jcp.25961   open full text
• Melatonin‐induced increase of lipid droplets accumulation and in vitro maturation in porcine oocytes is mediated by mitochondrial quiescence.
  Bin He, Chao Yin, Yabin Gong, Jie Liu, Huiduo Guo, Ruqian Zhao.
  Journal of Cellular Physiology. May 23, 2017
  Melatonin, the major pineal secretory product, has a significant impact on the female reproductive system. Recently, the beneficial effects of melatonin on mammalian oocyte maturation and embryonic development have drawn increased attention. However, the exact underlying mechanisms remain to be fully elucidated. This study demonstrates that supplementing melatonin to in vitro maturation (IVM) medium enhances IVM rate, lipid droplets (LDs) accumulation as well as triglyceride content in porcine oocytes. Decrease of mitochondrial membrane potential, mitochondrial respiratory chain complex IV activity as well as mitochondrial reactive oxygen species (mROS) content indicated that melatonin induced a decrease of mitochondrial activity. The copy number of mitochondrial DNA (mtDNA) which encodes essential subunits of oxidative phosphorylation (OXPHOS), was not affected by melatonin. However, the expression of mtDNA‐encoded genes was significantly down‐regulated after melatonin treatment. The DNA methyltransferase DNMT1, which regulates methylation and expression of mtDNA, was increased and translocated into the mitochondria in melatonin‐treated oocytes. The inhibitory effect of melatonin on the expression of mtDNA was significantly prevented by simultaneous addition of DNMT1 inhibitor, which suggests that melatonin regulates the transcription of mtDNA through up‐regulation of DNMT1 and mtDNA methylation. Increase of triglyceride contents after inhibition of OXPHOS indicated that mitochondrial quiescence is crucial for LDs accumulation in oocytes. Taken together, our results suggest that melatonin‐induced reduction in mROS production and increase in IVM, and LDs accumulation in porcine oocytes is mediated by mitochondrial quiescence.

  May 23, 2017   doi: 10.1002/jcp.25876   open full text
• NLRP3 inflammasome: Its regulation and involvement in atherosclerosis.
  Zahra Hoseini, Fatemeh Sepahvand, Bahman Rashidi, Amirhossein Sahebkar, Aria Masoudifar, Hamed Mirzaei.
  Journal of Cellular Physiology. May 23, 2017
  Inflammasomes are intracellular complexes involved in the innate immunity that convert proIL‐1β and proIL‐18 to mature forms and initiate pyroptosis via cleaving procaspase‐1. The most well‐known inflammasome is NLRP3. Several studies have indicated a decisive and important role of NLRP3 inflammasome, IL‐1β, IL‐18, and pyroptosis in atherosclerosis. Modern hypotheses introduce atherosclerosis as an inflammatory/lipid‐based disease and NLRP3 inflammasome has been considered as a link between lipid metabolism and inflammation because crystalline cholesterol and oxidized low‐density lipoprotein (oxLDL) (two abundant components in atherosclerotic plaques) activate NLRP3 inflammasome. In addition, oxidative stress, mitochondrial dysfunction, endoplasmic reticulum (ER) stress, and lysosome rupture, which are implicated in inflammasome activation, have been discussed as important events in atherosclerosis. In spite of these clues, some studies have reported that NLRP3 inflammasome has no significant effect in atherogenesis. Our review reveals that some molecules such as JNK‐1 and ASK‐1 (upstream regulators of inflammasome activation) can reduce atherosclerosis through inducing apoptosis in macrophages. Notably, NLRP3 inflammasome can also cause apoptosis in macrophages, suggesting that NLRP3 inflammasome may mediate JNK‐induced apoptosis, and the apoptotic function of NLRP3 inflammasome may be a reason for the conflicting results reported. The present review shows that the role of NLRP3 in atherogenesis can be significant. Here, the molecular pathways of NLRP3 inflammasome activation and the implications of this activation in atherosclerosis are explained.

  May 23, 2017   doi: 10.1002/jcp.25930   open full text
• Targeting RAS signaling pathway as a potential therapeutic target in the treatment of colorectal cancer.
  Afsane Bahrami, Seyed Mahdi Hassanian, Soodabeh ShahidSales, Zahra Farjami, Malihe Hasanzadeh, Kazem Anvari, Amir Aledavood, Mina Maftouh, Gordon A. Ferns, Majid Khazaei, Amir Avan.
  Journal of Cellular Physiology. May 23, 2017
  The V‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) is frequently dysregulated in colorectal cancer (CRC). It is involved in the modulation of several downstream effectors, that include: Raf/Mek/Erk, PI3K/Akt, RalGDS/p38MAPK, and Rac/Rho, and thereby influences tumorigenesis, the invasive behaviors of tumor cell, and resistance to therapy. There is growing evidence exploring the use of drugs that target these pathways in the treatment of CRC. Cetuximab has been approved for CRC patients without a KRAS mutation, or for EGFR‐expressing metastatic CRC, although some of the patients have a mutation of KRAS and NRAS. This review summarizes the recent knowledge about the therapeutic potential of targeting RAS with particular emphasis on recent preclinical and clinical studies in treatment of CRC.

  May 23, 2017   doi: 10.1002/jcp.25890   open full text
• Fibroblast dynamics as an in vitro screening platform for anti‐fibrotic drugs in primary myelofibrosis.
  Ciprian Tomuleasa, Sonia Selicean, Grigore Gafencu, Bobe Petrushev, Laura Pop, Cristian Berce, Anca Jurj, Adrian Trifa, Ana‐Maria Rosu, Sergiu Pasca, Lorand Magdo, Mihnea Zdrenghea, Delia Dima, Alina Tanase, Ioana Frinc, Anca Bojan, Ioana Berindan‐Neagoe, Gabriel Ghiaur, Stefan O. Ciurea.
  Journal of Cellular Physiology. May 23, 2017
  Although the cause for bone marrow fibrosis in patients with myelofibrosis remains controversial, it has been hypothesized that it is caused by extensive fibroblast proliferation under the influence of cytokines generated by the malignant megakaryocytes. Moreover, there is no known drug therapy which could reverse the process. We studied the fibroblasts in a novel system using the hanging drop method, evaluated whether the fibroblasts obtain from patients are part of the malignant clone of not and, using this system, we screen a large library of FDA‐approved drugs to identify potential drugs candidates that might be useful in the treatment of this disease, specifically which would inhibit fibroblast proliferation and the development of bone marrow fibrosis. We have found that the BM fibroblasts are not part of the malignant clone, as previously suspected and two immunosuppressive medications—cyclosporine and mycophenolate mophetil, as most potent suppressors of the fibroblast collagen production thus potentially inhibitors of bone marrow fibrosis production in myelofibrosis. Although the cause for bone marrow fibrosis in patients with myelofibrosis remains controversial, it has been hypothesized that it is caused by extensive fibroblast proliferation under the influence of cytokines generated by the malignant megakaryocytes. Moreover, there is no known drug therapy which could reverse the process. We studied the fibroblasts in a novel system using the hanging drop method, evaluated whether the fibroblasts obtain from patients are part of the malignant clone of not and, using this system, we screen a large library of FDA‐approved drugs to identify potential drugs candidates that might be useful in the treatment of this disease, specifically which would inhibit fibroblast proliferation and the development of bone marrow fibrosis. We have found that the BM fibroblasts are not part of the malignant clone, as previously suspected and two immunosuppressive medications—cyclosporine and mycophenolate mophetil, as most potent suppressors of the fibroblast collagen production thus potentially inhibitors of bone marrow fibrosis production in myelofibrosis.

  May 23, 2017   doi: 10.1002/jcp.25902   open full text
• Genetic variants as potential predictive biomarkers in advanced colorectal cancer patients treated with oxaliplatin‐based chemotherapy.
  Afsane Bahrami, Forouzan Amerizadeh, Seyed Mahdi Hassanian, Soodabeh ShahidSales, Majid Khazaei, Mina Maftouh, Majid Ghayour‐Mobarhan, Gordon A. Ferns, Amir Avan.
  Journal of Cellular Physiology. May 23, 2017
  Chemotherapy regimen containing oxaliplatin is often the first‐line treatment for patient with advanced colorectal cancer. Oxaliplatin binds to DNA, leading to the formation of crosslinks and bulky adducts. Approximately 50% of patients with CRC benefit from treatment with oxaliplatin. It is possible that genetic variants in biological pathways involved in drug transportation, drug metabolism, DNA damage repair, and cell cycle modulation might affect the activity, or efficacy of oxaliplatin. Because oxaliplatin resistance may be related to these genetic variants and may therefore be an important reason for treatment failure, we have summarized the genetic variations that have been reported to be predictive markers of the response to oxaliplatin based therapy in patients with advanced CRC.

  May 23, 2017   doi: 10.1002/jcp.25966   open full text
• GD2‐targeted immunotherapy and potential value of circulating microRNAs in neuroblastoma.
  Sharareh Gholamin, Hamed Mirzaei, Seyed‐Mostafa Razavi, Seyed Mahdi Hassanian, Leila Saadatpour, Aria Masoudifar, Soodabeh ShahidSales, Amir Avan.
  Journal of Cellular Physiology. May 23, 2017
  Neuroblastoma (NB) with various clinical presentation is a known childhood malignancy. Despite significant progress in treatment of NB afflicted patients, high risk disease is usually associated with poor outcome, resulting in long‐term survival of less that 50%. Known as a disease most commonly originated form the nerve roots, the variants involved in NB imitation and progression remain to be elucidated. The outcome of low to intermediate risk disease is favorable whereas the high risk NB disease with dismal prognosis, positing the necessity of novel approaches for early detection and prognostication of advanced disease. Tailored immunotherapy approaches have shown significant improvement in high‐risk NB patients. It has found a link between Gangliosides and progression of NB. The vast majority of neuroblastoma tumors express elevated levels of GD2, opening new insight into using anti‐GD2 drugs as potential treatments for NBs. Implication of anti‐GD2 monoclonal antibodies for treatment of high risk NBs triggers further investigation to unearth novel biomarkers as prognostic and response biomarker to guide additional multimodal tailored treatment approaches. A growing body of evidence supports the usefulness of miRNAs to evaluate high risk NBs response to anti‐GD2 drugs and further prevent drug‐related toxicities in refractory or recurrent NBs. miRNAs and circulating proteins in body fluids (plasma and serum) present as potential biomarkers in early detection of NBs. Here, we summarize various biomarkers involved in diagnosis, prognosis and response to treatment in patients with NB. We further attempted to overview prognostic biomarkers in response to treatment with anti‐GD2 drugs.

  May 23, 2017   doi: 10.1002/jcp.25793   open full text
• The therapeutic potential of targeting the BRAF mutation in patients with colorectal cancer.
  Afsane Bahrami, AmirReza Hesari, Majid Khazaei, Seyed Mahdi Hassanian, Gordon A. Ferns, Amir Avan.
  Journal of Cellular Physiology. May 23, 2017
  Colorectal cancer is among the most lethal malignancies globally. BRAF is a member of the RAS/RAF/MEK/ERK signaling pathway. Its constitutive activation can result in increased cellular growth, development, invasion, and resistance to therapy. A mutation of the BRAF gene is present in 5–10% of metastatic colorectal cancers. BRAF mutations have been found to predict a lack of benefit to anti‐EGFR therapy in metastatic CRC. Furthermore, CRC containing the BRAF V600E mutation display an innate resistance to BRAF inhibitors. The mechanisms of cell resistance can be explained at least in part by ERK dependent and ERK in‐dependent pathway. Clinical trials evaluating the combinations of BRAF, PI3K, EGFR, and/or MEK inhibitors have revealed promising activity in BRAF mutant containing CRCs. There may be some benefit from future studies that focus on improving the efficacy of combined therapy in CRC with respect to the sustained effects. The aim of current review is to give an overview about the current status and prospective regarding the therapeutic potential of targeting BRAF mutant colorectal cancer.

  May 23, 2017   doi: 10.1002/jcp.25952   open full text
• Roles of neurotrophins in skeletal tissue formation and healing.
  Yu‐Wen Su, Xin‐Fu Zhou, Bruce K. Foster, Brian L. Grills, Jiake Xu, Cory J. Xian.
  Journal of Cellular Physiology. May 23, 2017
  Neurotrophins and their receptors are key molecules that are known to be critical in regulating nervous system development and maintenance and have been recognized to be also involved in regulating tissue formation and healing in skeletal tissues. Studies have shown that neurotrophins and their receptors are widely expressed in skeletal tissues, implicated in chondrogenesis, osteoblastogenesis, and osteoclastogenesis, and are also involved in regulating tissue formation and healing events in skeletal tissue. Increased mRNA expression for neurotrophins NGF, BDNF, NT‐3, and NT‐4, and their Trk receptors has been observed in injured bone tissues, and NT‐3 and its receptor, TrkC, have been identified to have the highest induction at the injury site in a drill‐hole injury repair model in both bone and the growth plate. In addition, NT‐3 has also recently been shown to be both an osteogenic and angiogenic factor, and this neurotrophin can also enhance expression of the key osteogenic factor, BMP‐2, as well as the major angiogenic factor, VEGF, to promote bone formation, vascularization, and healing of the injury site. Further studies, however, are needed to investigate if different neurotrophins have differential roles in skeletal repair, and if NT‐3 can be a potential target of intervention for promoting bone fracture healing. Increasing evidence now suggests that neurotrophins also have important roles in bone. Among the neurotrophins and receptors, the NT‐3/TrkC signaling (induced far more prominently compared to other NTs and other Trk receptors) may potentially play a prominent role in the bone healing.

  May 23, 2017   doi: 10.1002/jcp.25936   open full text
• Connexin43 intercellular communication drives the early differentiation of human bone marrow stromal cells into osteoblasts.
  Julie Talbot, Régis Brion, Audrey Lamora, Mathilde Mullard, Sarah Morice, Dominique Heymann, Franck Verrecchia.
  Journal of Cellular Physiology. May 23, 2017
  Although it has been demonstrated that human bone marrow stromal cells (hBMSCs) express the ubiquitous connexin43 (Cx43) and form functional gap junctions, their role in the early differentiation of hBMSCs into osteoblasts remains poorly documented. Using in vitro assays, we show that Cx43 expression and gap junctional intercellular communication (GJIC) are increased during the differentiation of hBMSCs into osteoblasts, both at the protein and mRNA levels. Two independent procedures to reduce GJIC, a pharmacological approach with GJIC inhibitors (18α‐glycyrrhetinic acid and Gap27 peptide) and a molecular approach using small interfering RNA against Cx43, demonstrated that the presence of Cx43 and functional junctional channels are essential to the ability of hBMSCs to differentiate into osteoblasts in vitro. In addition, a reduced GJIC decreases the expression of Runx2, the major transcription factor implicated in the control of osteoblast commitment and early differentiation of hBMSCs into osteoblasts, suggesting that GJIC mediated by Cx43 is implicated in this process. Together our results demonstrate that GJIC mediated by the Cx43 channels plays a central role throughout the differentiation of hBMSC into osteoblasts, from the early stages to the process of mineralization. Our results demonstrate that GJIC mediated by the Cx43 channels plays a central role throughout the differentiation of hBMSC into osteoblasts, from the early stages to the process of mineralization.

  May 23, 2017   doi: 10.1002/jcp.25938   open full text
• TWEAK promotes migration and invasion in MEFs through a mechanism dependent on ERKs activation and Fibulin 3 down‐regulation.
  Celia Sequera, Ana Vázquez‐Carballo, María Arechederra, Sonia Fernández‐Veledo, Almudena Porras.
  Journal of Cellular Physiology. May 23, 2017
  TWEAK regulates multiple physio‐pathological processes in fibroblasts such as fibrosis. It also induces migration and invasion in tumors and it can activate p38 MAPK in various cell types. Moreover, p38α MAPK promotes migration and invasion in several cancer cells types and in mouse embryonic fibroblasts (MEFs). However, it remains unknown if TWEAK could promote migration in fibroblasts and whether p38α MAPK might play a role. Our results reveal that TWEAK activates ERKs, Akt, and p38α/β MAPKs and reduces secreted Fibulin 3 in MEFs. TWEAK also increases migration and invasion in wt and p38α deficient MEFs, which indicates that p38α MAPK is not required to mediate these effects. In contrast, ERKs inhibition significantly decreases TWEAK‐induced migration and Fibulin 3 knock‐down mimics TWEAK effect. These results indicate that both ERKs activation and Fibulin 3 down‐regulation would contribute to mediate TWEAK pro‐migratory effect. In fact, the additional regulation of ERKs and/or p38β as a consequence of Fibulin 3 decrease might be also involved in the pro‐migratory effect of TWEAK in MEFs. In conclusion, our studies uncover novel mechanisms by which TWEAK would favor tissue repair by promoting fibroblasts migration. TWEAK promotes migration of mouse embryonic fibroblasts through ERKs activation and Fibulin 3 down‐regulation. The additional regulation of ERKs and/or p38beta as a consequence of Fibulin 3 decrease might be also involved in this pro‐migratory effect of TWEAK.

  May 23, 2017   doi: 10.1002/jcp.25942   open full text
• Identification and characterization of site‐specific N‐glycosylation in the potassium channel Kv3.1b.
  Paul Christian Vicente, Jin Young Kim, Jeong‐Ju Ha, Min‐Young Song, Hyun‐Kyung Lee, Dong‐Hyun Kim, Jin‐Sung Choi, Kang‐Sik Park.
  Journal of Cellular Physiology. May 19, 2017
  The potassium ion channel Kv3.1b is a member of a family of voltage‐gated ion channels that are glycosylated in their mature form. In the present study, we demonstrate the impact of N‐glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. Large quantities of asparagine 229 (N229)‐glycosylated Kv3.1b reached the plasma membrane, whereas N220‐glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). These ER‐retained Kv3.1b proteins were susceptible to degradation, when co‐expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. Mass spectrometry analysis revealed a complex type Hex3HexNAc4Fuc1 glycan as the major glycan component of the N229‐glycosylated Kv3.1b protein, as opposed to a high‐mannose type Man8GlcNAc2 glycan for N220‐glycosylated Kv3.1b. Taken together, these results suggest that trafficking‐dependent roles of the Kv3.1b potassium channel are dependent on N229 site‐specific glycosylation and N‐glycan structure, and operate through a mechanism whereby specific N‐glycan structures regulate cell surface expression. Site‐specific N‐glycosylation and N‐glycan structure regulate cell surface trafficking and expression of the Kv3.1b potassium channel. Mass spectrometry analysis identified Man8GlcNAc2 (high‐mannose type) and Hex3HexNAc4Fuc1 (complex type) as the glycan moieties on N220 and N229.

  May 19, 2017   doi: 10.1002/jcp.25915   open full text
• Transplantation of mesenchymal stem cells overexpressing IL10 attenuates cardiac impairments in rats with myocardial infarction.
  Xin Meng, Jianping Li, Ming Yu, Jian Yang, Minjuan Zheng, Jinzhou Zhang, Chao Sun, Hongliang Liang, Liwen Liu.
  Journal of Cellular Physiology. May 19, 2017
  Mesenchymal stem cell (MSC) has been well known to exert therapeutic potential for patients with myocardial infarction (MI). In addition, interleukin‐10 (IL10) could attenuate MI through suppressing inflammation. Thus, the combination of MSC implantation with IL10 delivery may extend health benefits to ameliorate cardiac injury after MI. Here we established overexpression of IL10 in bone marrow‐derived MSC through adenoviral transduction. Cell viability, apoptosis, and IL10 secretion under ischemic challenge in vitro were examined. In addition, MSC was transplanted into the injured hearts in a rat model of MI. Four weeks after the MI induction, MI, cardiac functions, apoptotic cells, and inflammation cytokines were assessed. In response to in vitro oxygen‐glucose deprivation (OGD), IL10 overexpression in MSC (Ad.IL10‐MSC) enhanced cell viability, decreased apoptosis, and increased IL10 secretion. Consistently, the implantation of Ad.IL10‐MSCs into MI animals resulted in more reductions in myocardial infarct size, cardiac impairment, and cell apoptosis, compared to the individual treatments of either MSC or IL10 administration. Moreover, the attenuation of both systemic and local inflammations was most prominent for Ad.IL10‐MSC treatment. IL10 overexpression and MSC may exert a synergistic anti‐inflammatory effect to alleviate cardiac injury after MI. IL10 overexpression in mesenchymal stem cells enhances cell viability and decreases apoptosis. Implantation of IL10‐overexpressed mesenchymal stem cells into myocardial infarct animals results in more reductions in myocardial infarct size, cardiac impairment, and cell apoptosis.

  May 19, 2017   doi: 10.1002/jcp.25919   open full text
• All‐trans retinoic acid ameliorates hepatic stellate cell activation via suppression of thioredoxin interacting protein expression.
  Hiroki Shimizu, Toshiaki Tsubota, Keita Kanki, Goshi Shiota.
  Journal of Cellular Physiology. May 19, 2017
  Activation of hepatic stellate cells (HSCs) is the effector factor of hepatic fibrosis and hepatocellular carcinoma (HCC) development. Accumulating evidence suggests that retinoic acids (RAs), derivatives of vitamin A, contribute to prevention of liver fibrosis and carcinogenesis, however, regulatory mechanisms of RAs still remain exclusive. To elucidate RA signaling pathway, we previously performed a genome‐wide screening of RA‐responsive genes by in silico analysis of RA‐response elements, and identified 26 RA‐responsive genes. We found that thioredoxin interacting protein (TXNIP), which inhibits antioxidant activity of thioredoxin (TRX), was downregulated by all‐trans retinoic acid (ATRA). In the present study, we demonstrate that ATRA ameliorates activation of HSCs through TXNIP suppression. HSC activation was attenuated by TXNIP downregulation, whereas potentiated by TXNIP upregulation, indicating that TXNIP plays a crucial role in activation of HSCs. Notably, we showed that TXNIP‐mediated HSC activation was suppressed by antioxidant N‐acetylcysteine. In addition, ATRA treatment or downregulation of TXNIP clearly declined oxidative stress levels in activated HSCs. These data suggest that ATRA plays a key role in inhibition of HSC activation via suppressing TXNIP expression, which reduces oxidative stress levels. ATRA plays a key role in inhibition of HSC activation via suppressing TXNIP expression, which reduces oxidative stress levels.

  May 19, 2017   doi: 10.1002/jcp.25921   open full text
• The role of Runx2 in facilitating autophagy in metastatic breast cancer cells.
  Manish Tandon, Ahmad H. Othman, Vivek Ashok, Gary S. Stein, Jitesh Pratap.
  Journal of Cellular Physiology. May 19, 2017
  Breast cancer metastases cause significant patient mortality. During metastases, cancer cells use autophagy, a catabolic process to recycle nutrients via lysosomal degradation, to overcome nutritional stress for their survival. The Runt‐related transcription factor, Runx2, promotes cell survival under metabolic stress, and regulates breast cancer progression and bone metastases. Here, we identify that Runx2 enhances autophagy in metastatic breast cancer cells. We defined Runx2 function in cellular autophagy by monitoring microtubule‐associated protein light chain (LC3B‐II) levels, an autophagy‐specific marker. The electron and confocal microscopic analyses were utilized to identify alterations in autophagic vesicles. The Runx2 knockdown cells accumulate LC3B‐II protein and autophagic vesicles due to reduced turnover. Interestingly, Runx2 promotes autophagy by enhancing trafficking of LC3B vesicles. Our mechanistic studies revealed that Runx2 promotes autophagy by increasing acetylation of α‐tubulin sub‐units of microtubules. Inhibiting autophagy decreased cell adhesion and survival of Runx2 knockdown cells. Furthermore, analysis of LC3B protein in clinical breast cancer specimens and tumor xenografts revealed significant association between high Runx2 and low LC3B protein levels. Our studies reveal a novel regulatory mechanism of autophagy via Runx2 and provide molecular insights into the role of autophagy in metastatic cancer cells. We demonstrate a novel function of Runx2 in facilitating autophagy by altering cytoskeletal elements during cellular stress conditions such as nutrient deprivation. Our results in bone metastatic MDA‐MB‐231 breast cancer cells show that Runx2 promotes autophagy via α‐tubulin acetylation and autophagosome trafficking. Since autophagy promotes tumor cell adhesion, the Runx2‐mediated autophagy supporting cell adhesion highlights its function in metastatic breast cancer cell survival.

  May 19, 2017   doi: 10.1002/jcp.25916   open full text
• Gax suppresses chemerin/CMKLR1‐induced preadipocyte biofunctions through the inhibition of Akt/mTOR and ERK signaling pathways.
  Yunqi Jiang, Ping Liu, Wenlin Jiao, Juan Meng, Jinbo Feng.
  Journal of Cellular Physiology. May 19, 2017
  Adipose tissue is closely associated with angiogenesis and vascular remodeling. Chemerin is involved in inflammatory reaction and vascular dysfunction. However, the mechanisms of chemerin participating in vascular remodeling and whether Growth arrest‐specific homeobox (Gax) can effectively intervene it remain obscured. Here, 3T3‐F442A preadipocytes were cultured, injected into athymic mice to model fat pads, and treated respectively with Ad‐chemerin, Ad‐Gax, or specific inhibitors in vitro and in vivo. MTT, flow cytometry, Western blotting, and imunohisto(cyto)‐chemistry analyses showed that chemerin enhanced the expression of FABP4 and VEGF, activated Akt/mTOR and ERK pathways, increased the cell percent of S phase, decreased the percent of G0‐G1 phase and apoptotic cells, and augmented neovascular density in fat pads. Inversely, Gax suppressed the expression of these adipogenic and vasifactive markers and these signaling proteins, decreased the percent of S phase cells, and increased those of G0‐G1 phase and apoptotic cells, and reduced the neovascular density. Our results indicate that chemerin‐CMKLR1 activates Akt/mTOR and ERK pathways and facilitates preadipocyte proliferation, adipogenesis, and angiogenesis. Contrarily, Gax weakens the effect of chemerin on preadipocyte biofunctions. Chemerin/CMKLR1 activates Akt/mTOR/p70SK1/4EBP1 and ERK1/2 pathways and enhances preadipocyte proliferation, adipogenesis, and angiogenesis. Contrarily, Gax weakens the effect of chemerin on the preadipocyte biofunctions.

  May 19, 2017   doi: 10.1002/jcp.25918   open full text
• Global chondrocyte gene expression after a single anabolic loading period: Time evolution and re‐inducibility of mechano‐responses.
  Simone Scholtes, Elisabeth Krämer, Melanie Weisser, Wolfgang Roth, Reto Luginbühl, Tobias Grossner, Wiltrud Richter.
  Journal of Cellular Physiology. May 19, 2017
  Aim of this study was a genome‐wide identification of mechano‐regulated genes and candidate pathways in human chondrocytes subjected to a single anabolic loading episode and characterization of time evolution and re‐inducibility of the response. Osteochondral constructs consisting of a chondrocyte‐seeded collagen‐scaffold connected to β‐tricalcium‐phosphate were pre‐cultured for 35 days and subjected to dynamic compression (25% strain, 1 Hz, 9 × 10 min over 3 hr) before microarray‐profiling was performed. Proteoglycan synthesis was determined by 35S‐sulfate‐incorporation over 24 hr. Cell viability and hardness of constructs were unaltered by dynamic compression while proteoglycan synthesis was significantly stimulated (1.45‐fold, p = 0.016). Among 115 significantly regulated genes, 114 were up‐regulated, 48 of them ≥ twofold. AP‐1‐relevant transcription factors FOSB and FOS strongly increased in line with elevated ERK1/2‐phosphorylation and rising MAP3K4 expression. Expression of proteoglycan‐synthesizing enzymes CHSY1 and GALNT4 was load‐responsive as were factors associated with the MAPK‐, TGF‐β‐, calcium‐, retinoic‐acid‐, Wnt‐, and Notch‐signaling pathway which were significantly upregulated SOX9, and BMP6 levels rose significantly also after multiple loading episodes at daily intervals even at the 14th cycle with no indication for desensitation. Canonical pSmad2/3 and pSmad1/5/9‐signaling showed no consistent regulation. This study associates novel genes with mechanoregulation in chondrocytes, raising SOX9 protein levels with anabolic loading and suggests that more pathways than so far anticipated apparently work together in a complex network of stimulators and feedback‐regulators. Upregulation of mechanosensitive indicators extending differentially into the resting time provides crucial knowledge to maximize cartilage matrix deposition for the generation of high‐level cartilage replacement tissue. We here identified mechano‐regulation of SOX9 in chondrocytes in the center of a complex signaling network of an anabolic loading context and describe its robust re‐inducibility in daily intervals without desensitization. We suggest to use SOX9 protein levels and a promising selection of critical mechanosensitive genes as diagnostic markers to optimize loading regimes for future refinement of anabolic loading protocols to enhance cartilage tissue functionality in vitro and of tissue formation after surgical therapy in patients in vivo.

  May 19, 2017   doi: 10.1002/jcp.25933   open full text
• Acute ethanol exposure‐induced autophagy‐mediated cardiac injury via activation of the ROS‐JNK‐Bcl‐2 pathway.
  Zhongxin Zhu, Yewei Huang, Lingchun Lv, Youli Tao, Minglong Shao, Congcong Zhao, Mei Xue, Jia Sun, Chao Niu, Yang Wang, Sunam Kim, Weitao Cong, Wei Mao, Litai Jin.
  Journal of Cellular Physiology. May 19, 2017
  Binge drinking is associated with increased cardiac autophagy, and often triggers heart injury. Given the essential role of autophagy in various cardiac diseases, this study was designed to investigate the role of autophagy in ethanol‐induced cardiac injury and the underlying mechanism. Our study showed that ethanol exposure enhanced the levels of LC3‐II and LC3‐II positive puncta and promoted cardiomyocyte apoptosis in vivo and in vitro. In addition, we found that ethanol induced autophagy and cardiac injury largely via the sequential triggering of reactive oxygen species (ROS) accumulation, activation of c‐Jun NH2‐terminal kinase (JNK), phosphorylation of Bcl‐2, and dissociation of the Beclin 1/Bcl‐2 complex. By contrast, inhibition of ethanol‐induced autophagic flux with pharmacologic agents in the hearts of mice and cultured cells significantly alleviated ethanol‐induced cardiomyocyte apoptosis and heart injury. Elimination of ROS with the antioxidant N‐acetyl cysteine (NAC) or inhibition of JNK with the JNK inhibitor SP600125 reduced ethanol‐induced autophagy and subsequent autophagy‐mediated apoptosis. Moreover, metallothionein (MT), which can scavenge reactive oxygen and nitrogen species, also attenuated ethanol‐induced autophagy and cell apoptosis in MT‐TG mice. In conclusion, our findings suggest that acute ethanol exposure induced autophagy‐mediated heart toxicity and injury mainly through the ROS‐JNK‐Bcl‐2 signaling pathway. Acute ethanol‐induces cardiac dysfunction and cardiomyocyte apoptosis through upregulated autophagy. Acute ethanol modulation of autophagic activity may be mediated by the ROS‐JNK‐Bcl‐2 signaling pathway.

  May 19, 2017   doi: 10.1002/jcp.25934   open full text
• Autophagy maintains the integrity of endothelial barrier in LPS‐induced lung injury.
  Dan Zhang, Jian Zhou, Le Chi Ye, Jing Li, Zhenzhou Wu, Yuping Li, Chichi Li.
  Journal of Cellular Physiology. May 19, 2017
  Understanding the role and underlying regulation mechanism of autophagy in lipopolysaccharide‐induced lung injury (LPS‐LI) may provide potentially new pharmacological targets for treatment of acute lung injury. The aim of this study was to investigate the functional significance of autophagy in LPS‐LI. The autophagy of human pulmonary microvascular endothelial cells (HPMVECs) and mice was inhibited before they were challenged with LPS. In vitro, permeability, vitality, and the LDH release rate of the cells were detected, the zonula occluden‐1 (ZO‐1) expression and the stress fiber formation were determined. In vivo, the lung injury was assessed. We found LPS caused high permeability and increased lactate dehydrogenase (LDH) release rate, lowered viability of the cells, inhibited the ZO‐1 expression and induced stress fiber formation, these effects were further aggravated by prohibiting the level of autophagy. Consistently, in in vivo experiments, LPS‐induced serious lung injury, which was reflected as edema, leukocyte infiltration and hemorrhage in lung tissue, and the high concentration of pro‐inflammation cytokines tumor necrosis factor (TNF)‐α and interleukin (IL)‐1β in bronchoalveolar lavage fluid (BALF). Inhibiting autophagy further exacerbated LPS‐LI. It appears that autophagy played a protective role in LPS‐LI in part through restricting the injury of lung microvascular barrier.

  May 19, 2017   doi: 10.1002/jcp.25928   open full text
• Mechanism of prolactin inhibition of miR‐135b via methylation in goat mammary epithelial cells.
  Zhi Chen, Jun Luo, ChangHui Zhang, Yue Ma, Shuang Sun, Tianyin Zhang, Juan J. Loor.
  Journal of Cellular Physiology. May 19, 2017
  Prolactin is an important endocrine activator of lactogenesis. This study investigated the function and mechanism of miR‐135b in the enhancement of lactation by prolactin in goat mammary epithelial tissue. We utilized S‐Poly (T) sequencing to evaluate changes in gene regulation in the goat mammary gland after incubation with 2.5 μg/ml prolactin and 2.5 μg/ml IGF‐1 by examining highly expressed miRNAs during early lactation and late‐lactation. The results illustrated that miR‐135b is highly expressed in the goat mammary gland during early lactation and late‐lactation, and also after treatment with 2.5 μg/ml prolactin and 2.5 μg/ml IGF‐1. We used Q‐RT PCR, Western Blot, immunofluorescence, and luciferase reporter assay analysis, and found that PRL was significantly down‐regulated in response to the expression of miR‐135b in a manner that was functionally related to TAG synthesis via the large tumor suppressor 2 gene (Lats2), an important regulator of adipocyte proliferation via Hippo Signaling. Furthermore, using bisulfite‐sequencing PCR (BSP), Q‐PCR, and Western Blot we discovered an increase in expression of DNMT I (DNA methyl transferase I) in goat mammary epithelial cells with the 2.5 μg/ml PRL incubation, which led to DNA methylation of the CpG island upstream of miR‐135b and inhibited the transcription and expression of miR‐135b. This study clarified the mechanism by which PRL induces milk formation and identified a role of miR‐135b in this process. With PRL stimulation, the expression of DNMT I in mammary epithelial cells increased, resulting in DNA methylation of the CpG island upstream of miR‐135b's, thus inhibiting transcription of miR‐135b. At the same time, down‐regulation of miR‐135b resulted in increased expression of the target gene LATS2 thereby causing proliferation and metabolic changes in the mammary gland, eventually leading to lactation.

  May 19, 2017   doi: 10.1002/jcp.25925   open full text
• Primary cilium alterations and expression changes of Patched1 proteins in niemann‐pick type C disease.
  Patrizia Formichi, Carla Battisti, Maria M. De Santi, Raffaella Guazzo, Sergio A. Tripodi, Elena Radi, Benedetta Rossi, Ermelinda Tarquini, Antonio Federico.
  Journal of Cellular Physiology. May 19, 2017
  Niemann‐Pick type C disease (NPC) is a disorder characterized by abnormal intracellular accumulation of unesterified cholesterol and glycolipids. Two distinct disease‐causing genes have been isolated, NPC1 and NPC2. The NPC1 protein is involved in the sorting and recycling of cholesterol and glycosphingolipids in the late endosomal/lysosomal system. It has extensive homology with the Patched1 (Ptc1) receptor, a transmembrane protein localized in the primary cilium, and involved in the Hedgehog signaling (Shh) pathway. We assessed the presence of NPC1 and Ptc1 proteins and evaluated the relative distribution and morphology of primary cilia in fibroblasts from five NPC1 patients and controls, and in normal fibroblasts treated with 3‐ß‐[2‐(diethylamino)ethoxy]androst‐5‐en‐17‐one (U18666A), a cholesterol transport‐inhibiting drug that is widely used to mimic NPC. Immunofluorescence and western blot analyses showed a significant decrease in expression of NPC1 and Ptc1 in NPC1 fibroblasts, while they were normally expressed in U18666A‐treated fibroblasts. Moreover, fibroblasts from NPC1 patients and U18666A‐treated cells showed a lower percentage distribution of primary cilia and a significant reduction in median cilia length with respect to controls. These are the first results demonstrating altered cytoplasmic expression of Ptc1 and reduced number and length of primary cilia, where Ptc1 is located, in fibroblasts from NPC1 patients. We suggest that the alterations in Ptc1 expression in cells from NPC1 patients are closely related to NPC1 expression deficit, while the primary cilia alterations observed in NPC1 and U18666A‐treated fibroblasts may represent a secondary event derived from a defective metabolic pathway. We assessed the presence of NPC1 and Ptc1 proteins and evaluated the relative distribution and morphology of primary cilia in fibroblasts from five NPC1 patients. We demonstrated altered cytoplasmic expression of Ptc1 and reduced number and length of primary cilia in fibroblasts from NPC1 patients.

  May 19, 2017   doi: 10.1002/jcp.25926   open full text
• Retinoic acid receptor‐related orphan receptor RORα regulates differentiation and survival of keratinocytes during hypoxia.
  Hongyu Li, Longjian Zhou, Jun Dai.
  Journal of Cellular Physiology. May 19, 2017
  Low O2 pressures present in the microenvironment of epidermis control keratinocyte differentiation and epidermal barrier function through hypoxia inducible factors (HIFs) dependent gene expression. This study focuses on investigating relations of the retinoic acid receptor‐related orphan receptor alpha (RORα) to HIF‐1α in keratinocytes under hypoxic conditions. The expression level of RORα is significantly elevated under hypoxia in both human and murine keratinocytes. Gene silencing of RORA attenuates hypoxia‐stimulated expression of genes related to late differentiation and epidermal barrier function, and leads to an enhanced apoptotic response. While the hypoxic induction of RORα is dependent on HIF‐1α, RORα is in turn critical for nuclear accumulation of HIF‐1α and activation of HIF transcriptional activity. These results collectively suggest that RORα functions as an important mediator of HIF‐1α activities in regulating keratinocyte differentiation/survival and epidermal barrier function during the oxygen sensing stage. We have identified RORα as a regulator imperative to HIF‐1α activities in promoting terminal differentiation, epidermal barrier function, and survival of keratinocytes under low O2 tension. Given highly diversified functions of HIF targets in maintaining skin homeostasis, manipulations of HIF activities through RORα ligands can represent a novel strategy for therapeutic treatment of pathophysiological conditions such as cutaneous diseases related to defective epidermal barrier functions and wound healing.

  May 19, 2017   doi: 10.1002/jcp.25924   open full text
• Involvement of testicular DAAM1 expression in zinc protection against cadmium‐induced male rat reproductive toxicity.
  Marouane Chemek, Massimo Venditti, Sana Boughamoura, Safa B. Mimouna, Imed Messaoudi, Sergio Minucci.
  Journal of Cellular Physiology. May 19, 2017
  In order to verify the effects of exposure to Cd and Zn on testicular DAAM1 gene and protein expression and also to ascertain their involvement in the protective role of Zn in prevent the testicular toxicity Cd‐induced in male offspring rats at adult age after gestational and lactational exposure, male offspring rats, from mothers receiving either tap water, Cd, Zn, or Cd + Zn during gestation and lactation periods, were scarified on postnatal days (PND) 70. The reproductive organ (testis, epididymis, and vesicle seminal) were collected, weighed, and analyzed. The results showed that exposure to Cd in utero and through lactation decreased the relative reproductive organ weight, altered the testicular histology at the interstitial and tubular levels, and causing a significant reduction in the daily sperm production (DSP) per testis and per gram of testis, and other then altering the epididymal sperm quality. Furthermore, both mRNA and protein expression of rat testicular DAAM1 were also inhibited in Cd‐treated group. Zn supply has completely corrected the most of these toxic effects. Our results imply that Zn could prevent Cd‐induced testicular toxicity and sperm quality alteration in adult male rat after gestational and lactational exposure, probably via the restoration of the testicular DAAM1 expression inhibited by Cd. The effects of exposure to Cd and Zn on testicular DAAM1 gene and protein expression and heir involvement in the protective role of Zn in prevent the testicular toxicity Cd‐induced in male offspring rats have been studied. The reproductive organ were collected, weighed and analyzed. The results showed that Cd exposure decreased the relative reproductive organ weight, altered the testicular histology, and causing a significant reduction in the daily sperm production other then altering the epididymal sperm quality. In addition, both mRNA and protein expression of rat testicular DAAM1 were also inhibited. Zn supply has completely corrected the most of these toxic effects. Our results imply that Zn could prevent Cd‐induced testicular toxicity probably via the restoration of the testicular DAAM1 expression inhibited by Cd.

  May 19, 2017   doi: 10.1002/jcp.25923   open full text
• Dual‐specificity tyrosine phosphorylation‐regulated kinase 2 regulates osteoclast fusion in a cell heterotypic manner.
  Gali Guterman‐Ram, Milena Pesic, Ayelet Orenbuch, Tal Czeiger, Anastasia Aflalo, Noam Levaot.
  Journal of Cellular Physiology. May 19, 2017
  Monocyte fusion into osteoclasts, bone resorbing cells, plays a key role in bone remodeling and homeostasis; therefore, aberrant cell fusion may be involved in a variety of debilitating bone diseases. Research in the last decade has led to the discovery of genes that regulate osteoclast fusion, but the basic molecular and cellular regulatory mechanisms underlying the fusion process are not completely understood. Here, we reveal a role for Dyrk2 in osteoclast fusion. We demonstrate that Dyrk2 down regulation promotes osteoclast fusion, whereas its overexpression inhibits fusion. Moreover, Dyrk2 also promotes the fusion of foreign‐body giant cells, indicating that Dyrk2 plays a more general role in cell fusion. In an earlier study, we showed that fusion is a cell heterotypic process initiated by fusion‐founder cells that fuse to fusion‐follower cells, the latter of which are unable to initiate fusion. Here, we show that Dyrk2 limits the expansion of multinucleated founder cells through the suppression of the fusion competency of follower cells. This study investigate the molecular mechanisms regulating osteoclast fusion. Osteoclast fusion involve a fusion initiation cell termed fusion founder which fuse to cells lacking fusion initiation abilities termed fusion followers. This study show for the fist time that the protein Dyrk2 inhibit fusion by limiting the distribution of fusion follower cells.

  May 19, 2017   doi: 10.1002/jcp.25922   open full text
• miR‐124 and miR‐9 mediated downregulation of HDAC5 promotes neurite development through activating MEF2C‐GPM6A pathway.
  Xi Gu, Congcong Fu, Lifang Lin, Shuhu Liu, Xiaohong Su, Aili Li, Qiaoqi Wu, Chunhong Jia, Peidong Zhang, Lu Chen, Xinhong Zhu, Xuemin Wang.
  Journal of Cellular Physiology. May 19, 2017
  The class IIa histone deacetylases (HDACs) play important roles in the central nervous system during diverse biological processes such as synaptic plasticity, axon regeneration, cell apoptosis, and neural differentiation. Although it is known that HDAC5 regulates neuronal differentiation, neither the physiological function nor the regulation of HDAC5 in neuronal differentiation is clear. Here, we identify HDAC5 as an inhibitor of neurite elongation and show that HDAC5 is regulated by the brain enriched microRNA miR‐124 and miR‐9. We discover that HDAC5 inhibits neurite extension both in differentiated P19 cells and primary neurons. We also show that the neuronal membrane glycoprotein GPM6A (M6a) is a direct target gene of HDAC5 regulated transcriptional factor MEF2C. HDAC5 inhibits neurite elongation, acting at least partially via a MEF2C/M6a signaling pathway. We also confirmed the miR‐124/miR‐9 regulated HDAC5‐MEF2C‐M6a pathway regulates neurite development in primary neurons. Thus, HDAC5 emerges as a cellular conductor of MEF2C and M6a activity and is regulated by miR‐124 and miR‐9 to control neurite development. We provide evidence that through the direct repression of HDAC5, miR‐124, and miR‐9 intrinsically regulate MEF2C transcription activity and M6a expression, and this ensures proper neurite elongation. Our findings further suggest that by acting in neurons, brain‐enriched miRNAs can play key roles in coordinating vertebrate central nervous system development.

  May 19, 2017   doi: 10.1002/jcp.25927   open full text
• Lipopolysaccharide can modify differentiation and immunomodulatory potential of periodontal ligament stem cells via ERK1,2 signaling.
  Tamara Kukolj, Drenka Trivanović, Ivana Okić Djordjević, Slavko Mojsilović, Jelena Krstić, Hristina Obradović, Srdja Janković, Juan Francisco Santibanez, Aleksandra Jauković, Diana Bugarski.
  Journal of Cellular Physiology. May 19, 2017
  Lipopolysaccharide (LPS) is a pertinent deleterious factor in oral microenvironment for cells which are carriers of regenerative processes. The aim of this study was to investigate the emerging in vitro effects of LPS (Escherichia coli) on human periodontal ligament stem cell (PDLSC) functions and associated signaling pathways. We demonstrated that LPS did not affect immunophenotype, proliferation, viability, and cell cycle of PDLSCs. However, LPS modified lineage commitment of PDLSCs inhibiting osteogenesis by downregulating Runx2, ALP, and Ocn mRNA expression, while stimulating chondrogenesis and adipogenesis by upregulating Sox9 and PPARγ mRNA expression. LPS promoted myofibroblast‐like phenotype of PDLSCs, since it significantly enhanced PDLSC contractility, as well as protein and/or gene expression of TGF‐β, fibronectin (FN), α‐SMA, and NG2. LPS also increased protein and gene expression levels of anti‐inflammatory COX‐2 and pro‐inflammatory IL‐6 molecules in PDLSCs. Inhibition of peripheral blood mononuclear cells (MNCs) transendothelial migration in presence of LPS‐treated PDLSCs was accompanied by the reduction of CD29 expression within MNCs. However, LPS treatment did not change the inhibitory effect of PDLSCs on mitogen‐stimulated proliferation of CD4+ and the ratio of CD4+CD25high/CD4+CD25low lymphocytes. LPS‐treated PDLSCs did not change the frequency of CD34+ and CD45+ cells, but decreased the frequency of CD33+ and CD14+ myeloid cells within MNCs. Moreover, LPS treatment attenuated the stimulatory effect of PDLSCs on CFC activity of MNCs, predominantly the CFU‐GM number. The results indicated that LPS‐activated ERK1,2 was at least partly involved in the observed effects on PDLSC differentiation capacity, acquisition of myofibroblastic attributes, and changes of their immunomodulatory features. In this study, we examined functional properties of human periodontal ligament stem cells (PDLSCs) in presence of LPS (E. coli). Our results indicated that LPS modified PDLSCs’ mesodermal lineage commitment, favoring myofibroblastic‐like phenotype, without affecting their growth and immunophenotype. LPS‐treated PDLSCs strongly inhibited transendothelial migration (TEM) of human peripheral blood mononuclear cells (MNCs). LPS‐treated PDLSCs decreased the frequency of CD33+ and CD14+ myeloid cells within MNCs. Moreover, LPS treatment attenuated the stimulatory effect of PDLSCs on CFC activity, predominantly the CFU‐GM number, of MNCs. Our results indicate that changes in differentiation and immunomodulatory properties of PDLSCs may be in partly governed by LPS‐activated ERK1,2 signaling cascade.

  May 19, 2017   doi: 10.1002/jcp.25904   open full text
• Comparative proteomic profiling of human osteoblast‐derived extracellular matrices identifies proteins involved in mesenchymal stromal cell osteogenic differentiation and mineralization.
  Marta Baroncelli, Bram C. van der Eerden, Yik‐Yang Kan, Rodrigo D. Alves, Jeroen A. Demmers, Jeroen van de Peppel, Johannes P. van Leeuwen.
  Journal of Cellular Physiology. May 19, 2017
  The extracellular matrix (ECM) is a dynamic component of tissue architecture that physically supports cells and actively influences their behavior. In the context of bone regeneration, cell‐secreted ECMs have become of interest as they reproduce tissue‐architecture and modulate the promising properties of mesenchymal stem cells (MSCs). We have previously created an in vitro model of human osteoblast‐derived devitalized ECM that was osteopromotive for MSCs. The aim of this study was to identify ECM regulatory proteins able to modulate MSC differentiation to broaden the spectrum of MSC clinical applications. To this end, we created two additional models of devitalized ECMs with different mineralization phenotypes. Our results showed that the ECM derived from osteoblast‐differentiated MSCs had increased osteogenic potential compared to ECM derived from undifferentiated MSCs and non‐ECM cultures. Proteomic analysis revealed that structural ECM proteins and ribosomal proteins were upregulated in the ECM from undifferentiated MSCs. A similar response profile was obtained by treating osteoblast‐differentiating MSCs with Activin‐A. Extracellular proteins were upregulated in Activin‐A ECM, whereas mitochondrial and membrane proteins were downregulated. In summary, this study illustrates that the composition of different MSC‐secreted ECMs is important to regulate the osteogenic differentiation of MSCs. These models of devitalized ECMs could be used to modulate MSC properties to regulate bone quality. In vitro cell‐derived extracellular matrices (ECM) mimic the ECM role in tissue architecture and in actively modulating cell behavior, and they have been proposed for tissue engineering applications. We created three models of human osteoblast‐derived ECM with extremely different mineralization phenotypes, to identify regulatory proteins to modulate mesenchymal stromal cell (MSC) differentiation. Comparative proteomic profiles of these ECMs reveal that the protein composition is important to regulate MSC behavior, and could be used to modulate MSC properties and bone quality for bone tissue engineering applications.

  May 19, 2017   doi: 10.1002/jcp.25898   open full text
• Nanotubes impregnated human olfactory bulb neural stem cells promote neuronal differentiation in trimethyltin‐induced neurodegeneration rat model.
  Hany E. Marei, Ahmed A. Elnegiry, Adel Zaghloul, Asma Althani, Nahla Afifi, Ahmed Abd‐Elmaksoud, Amany Farag, Samah Lashen, Shymaa Rezk, Zeinab Shouman, Carlo Cenciarelli, Anwarul Hasan.
  Journal of Cellular Physiology. May 18, 2017
  Neural stem cells (NSCs) are multipotent self‐renewing cells that could be used in cellular‐based therapy for a wide variety of neurodegenerative diseases including Alzheimer's diseases (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS). Being multipotent in nature, they are practically capable of giving rise to major cell types of the nervous tissue including neurons, astrocytes, and oligodendrocytes. This is in marked contrast to neural progenitor cells which are committed to a specific lineage fate. In previous studies, we have demonstrated the ability of NSCs isolated from human olfactory bulb (OB) to survive, proliferate, differentiate, and restore cognitive and motor deficits associated with AD, and PD rat models, respectively. The use of carbon nanotubes (CNTs) to enhance the survivability and differentiation potential of NSCs following their in vivo engraftment have been recently suggested. Here, in order to assess the ability of CNTs to enhance the therapeutic potential of human OBNSCs for restoring cognitive deficits and neurodegenerative lesions, we co‐engrafted CNTs and human OBNSCs in TMT‐neurodegeneration rat model. The present study revealed that engrafted human OBNSCS‐CNTs restored cognitive deficits, and neurodegenerative changes associated with TMT‐induced rat neurodegeneration model. Moreover, the CNTs seemed to provide a support for engrafted OBNSCs, with increasing their tendency to differentiate into neurons rather than into glia cells. The present study indicate the marked ability of CNTs to enhance the therapeutic potential of human OBNSCs which qualify this novel therapeutic paradigm as a promising candidate for cell‐based therapy of different neurodegenerative diseases. In order to assess the ability of CNTs to enhance the therapeutic potential of human OBNSCs for restoring cognitive deficits and neurodegenerative lesions, we co‐engrafted CNTs and human OBNSCs in TMT‐neurodegeneration rat model. The present study revealed that engrafted human OBNSCS‐CNTs restored cognitive deficits, and neurodegenerative changes associated with TMT‐induced rat neurodegeneration model. The present study indicate the marked ability of CNTs to enhance the therapeutic potential of human OBNSCs which qualify this novel therapeutic paradigm as a promising candidate for cell‐based therapy of different neurodegenerative diseases.

  May 18, 2017   doi: 10.1002/jcp.25826   open full text
• Identification of murine phosphodiesterase 5A isoforms and their functional characterization in HL‐1 cardiac cell line.
  Federica Campolo, Alessandra Zevini, Silvia Cardarelli, Lucia Monaco, Federica Barbagallo, Manuela Pellegrini, Marisa Cornacchione, Antonio Di Grazia, Valeria De Arcangelis, Daniele Gianfrilli, Mauro Giorgi, Andrea Lenzi, Andrea M. Isidori, Fabio Naro.
  Journal of Cellular Physiology. May 18, 2017
  Phosphodiesterase 5A (PDE5A) specifically degrades the ubiquitous second messenger cGMP and experimental and clinical data highlight its important role in cardiac diseases. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2, and mPde5a3). The predicted amino acidic sequences of the three murine isoforms are different in the N‐terminal regulatory domain. mPDE5A isoforms were transfected in HEK293T cells and they showed high affinity for cGMP and similar sensitivity to sildenafil inhibition. RT‐PCR analysis showed that mPde5a1, mPde5a2, and mPde5a3 had differential tissue distribution. In the adult heart, mPde5a1 and mPde5a2 were expressed at different levels whereas mPde5a3 was undetectable. Overexpression of mPDE5As induced an increase of HL‐1 number cells which progress into cell cycle. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL‐1 areas, and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms. Moreover, mPDE5A isoforms had differential subcellular localization: mPDE5A1 was mainly localized in the cytoplasm, mPDE5A2 and mPDE5A3 were also nuclear localized. These results demonstrate for the first time the existence of three PDE5A isoforms in mouse and highlight their potential role in the induction of hypertrophy. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2, and mPde5a3). Overexpression of mPDE5As induced cell cycle progression of HL‐1 cells. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL‐1 areas, and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms.

  May 18, 2017   doi: 10.1002/jcp.25880   open full text
• Transcription factor EGR1 promotes differentiation of bovine skeletal muscle satellite cells by regulating MyoG gene expression.
  WeiWei Zhang, HuiLi Tong, ZiHeng Zhang, ShuLi Shao, Dan Liu, ShuFeng Li, YunQin Yan.
  Journal of Cellular Physiology. May 18, 2017
  The transcription factor, early growth response 1 (EGR1), has important roles in various cell types in response to different stimuli. EGR1 is thought to be involved in differentiation of bovine skeletal muscle‐derived satellite cells (MDSCs); however, the precise effects of EGR1 on differentiation of MDSCs and its mechanism of action remain unknown. In the present study, a time course of EGR1 expression and the effects of EGR1 on MDSC differentiation were determined. The results demonstrated that the expression of EGR1 mRNA and protein increased significantly in differentiating MDSCs relative to that in proliferating cells. Over‐expression of the EGR1 gene in MDSCs promoted their differentiation and inhibited proliferation. Conversely, knock‐down of EGR1 inhibited differentiation of MDSCs and promoted their proliferation, indicating that EGR1 promotes MDSC differentiation. Moreover, over‐expression of EGR1 in MDSCs increased the expression of MyoG mRNA and protein, whereas its knock‐down had the opposite effect. Furthermore, ChIP‐PCR analyses demonstrated that EGR1 could bind directly to its putative binding site within the promoter region of MyoG, and determination of ERG1 subcellular localization in MDSCs demonstrated that it could relocate to the nucleus, indicating MyoG is likely an EGR1 target gene whose expression is positively regulated by this transcription factor. In conclusion, EGR1 can promote MDSC differentiation through positive regulation of MyoG gene expression. The transcription factor, EGR1, can promote differentiation of bovine skeletal muscle‐derived satellite cells. In addition, EGR1 can relocate to the nucleus and bind directly to the MyoG gene promoter, hence positively regulating MyoG gene expression. These data prove that EGR1 promotes MDSC differentiation by regulation of MyoG gene expression.

  May 18, 2017   doi: 10.1002/jcp.25883   open full text
• Loss of TET1 facilitates DLD1 colon cancer cell migration via H3K27me3‐mediated down‐regulation of E‐cadherin.
  Zhen Zhou, Hong‐Sheng Zhang, Yang Liu, Zhong‐Guo Zhang, Guang‐Yuan Du, Hu Li, Xiao‐Ying Yu, Ying‐Hui Huang.
  Journal of Cellular Physiology. May 17, 2017
  Epigenetic modifications such as histone modifications and cytosine hydroxymethylation are linked to tumorigenesis. Loss of 5‐hydroxymethylcytosine (5hmC) by ten‐eleven translocation 1 (TET1) down‐regulation facilitates tumor initiation and development. However, the mechanisms by which loss of TET1 knockdown promotes malignancy development remains unclear. Here, we report that TET1 knockdown induced epithelial‐mesenchymal transition (EMT) and increased cancer cell growth, migration and invasion in DLD1 cells. Loss of TET1 increased EZH2 expression and reduced UTX‐1 expression, thus increasing histone H3K27 tri‐methylation causing repression of the target gene E‐cadherin. Ectopic expression of the H3K27 demethylase UTX‐1 or EZH2 depletion both impeded EZH2 binding caused a loss of H3K27 methylation at epithelial gene E‐cadherin promoter, thereby suppressing EMT and tumor invasion in shTET1 cells. Conversely, UTX‐1 depletion and ectopic expression of EZH2 enhanced EMT and tumor metastasis in DLD1 cells. These findings provide insight into the regulation of TET1 and E‐cadherin and identify EZH2 as a critical mediator of E‐cadherin repression and tumor progression.This article is protected by copyright. All rights reserved

  May 17, 2017   doi: 10.1002/jcp.26012   open full text
• A systems approach to physiologic evolution: From micelles to consciousness.
  John S. Torday, William B. Miller.
  Journal of Cellular Physiology. May 16, 2017
  A systems approach to evolutionary biology offers the promise of an improved understanding of the fundamental principles of life through the effective integration of many biologic disciplines. It is presented that any critical integrative approach to evolutionary development involves a paradigmatic shift in perspective, more than just the engagement of a large number of disciplines. Critical to this differing viewpoint is the recognition that all biological processes originate from the unicellular state and remain permanently anchored to that phase throughout evolutionary development despite their macroscopic appearances. Multicellular eukaryotic development can, therefore, be viewed as a series of connected responses to epiphenomena that proceeds from that base in continuous iterative maintenance of collective cellular homeostatic equipoise juxtaposed against an ever‐changing and challenging environment. By following this trajectory of multicellular eukaryotic evolution from within unicellular First Principles of Physiology forward, the mechanistic nature of complex physiology can be identified through a step‐wise analysis of a continuous arc of vertebrate evolution based upon serial exaptations. There is no systematic way of understanding physiology at the present time. By reducing the process of physiologic evolution to an integrated series of cellular‐molecular steps from the unicellular state to complex traits, this goal has now been accomplished.

  May 16, 2017   doi: 10.1002/jcp.25820   open full text
• Promising anti‐tumor properties of bisdemethoxycurcumin: A naturally occurring curcumin analogue.
  Mahin Ramezani, Mahdi Hatamipour, Amirhosein Sahebkar.
  Journal of Cellular Physiology. May 16, 2017
  Curcuminoids are turmeric‐extracted phytochemicals with documented chemopreventive and anti‐tumor activities against several types of malignancies. Curcuminoids can modulate several molecular pathways and cellular targets involved in different stages of tumor initiation, growth, and metastasis. Bisdemethoxycurcumin (BDMC) is a minor constituent (approximately 3%) of curcuminoids that has been shown to be more stable than the other two main curcuminoids, that is, curcumin and demthoxycurcumin. Recent studies have revealed that BDMC has anti‐tumor effects exerted through a multimechanistic mode of action involving inhibition of cell proliferation, invasion and migration, metastasis and tumour growth, and induction of apoptotic death in cancer cells. The present review discusses the findings on the anti‐tumor effects of BDMC, underlying mechanisms, and the relevance of finding for translational studies in human. This review discusses the findings on the anti‐tumor effects of bisdemethoxycurcumin, underlying mechanisms, and the relevance of finding for translational studies in human.

  May 16, 2017   doi: 10.1002/jcp.25795   open full text
• The prognostic value of MGMT promoter methylation in glioblastoma: A meta‐analysis of clinical trials.
  Maryam Moradi Binabaj, Afsane Bahrami, Soodabeh ShahidSales, Marjan Joodi, Mona Joudi Mashhad, Seyed Mahdi Hassanian, Kazem Anvari, Amir Avan.
  Journal of Cellular Physiology. May 16, 2017
  The DNA repair protein O6‐Methylguanine‐DNA methyltransferase (MGMT) is suggested to be associated with resistance to alkylating agents such as Temozolomide which is being used in treatment of patients with glioblastoma (GBM). Therefore, we evaluated the associations between MGMT promoter methylation and prognosis of patients with glioblastoma (GBM). Data were extracted from publications in Embase, PubMed, and the Cochrane Library. Data on overall survival (OS), progression‐free survival (PFS), and MGMT methylation status were obtained and 4,097 subjects were enrolled. Data from 34 studies showed that MGMT methylated patients had better OS, compared to GBM unmethylated patients (pooled HRs, 0.494; 95%CI 0.412–0.591; p = 0.001). Meta‐analysis of 10 eligible studies reporting on PFS, demonstrated that MGMT promoter methylation was not significantly associated with better PFS (pooled HRs, 0.653; 95%CI 0.414–1.030; p = 0.067). GBM patients with MGMT methylation were associated with longer overall survival, although this effect was not detected for PFS. Moreover, we performed further analysis in patients underwent a comprehensive imaging evaluation. This data showed a significant association with better OS and PFS, although further studies are warranted to assess the value of emerging marker in prospective setting in patients with glioblastoma as a risk stratification biomarker in clinical management of the patients.

  May 16, 2017   doi: 10.1002/jcp.25896   open full text
• Myocardial proteases and cardiac remodeling.
  Sadaf Riaz, Asad Zeidan, Fatima Mraiche.
  Journal of Cellular Physiology. May 16, 2017
  Cardiac hypertrophy (CH), characterized by the enlargement of cardiomyocytes, fibrosis and apoptosis, is one of the leading causes of death worldwide. Despite the advances in cardiovascular research, there remains a need to further investigate the signaling pathways that mediate CH in order to identify novel therapeutic targets. One of the hallmarks of CH is the remodeling of the extracellular matrix (ECM). Multiple studies have shown an important role of cysteine proteases and matrix metalloproteinases (MMPs) in the remodeled heart. This review focuses on the role of cysteine cathepins and MMPs in cardiac remodeling.

  May 16, 2017   doi: 10.1002/jcp.25884   open full text
• Intracellular transcytosis of albumin in glomerular endothelial cells after endocytosis through caveolae.
  Takahito Moriyama, Kayo Sasaki, Kazunori Karasawa, Keiko Uchida, Kosaku Nitta.
  Journal of Cellular Physiology. May 15, 2017
  We previously described albumin endocytosis through caveolae in human renal glomerular endothelial cells (HRGECs). This suggested a new albumin transcytosis pathway, in addition to the fenestral pathway. As a next step, we investigated albumin transcytosis in HRGECs after caveolar endocytosis. HRGECs were incubated with Alexa Fluor 488‐labeled bovine serum albumin from 0 to 360 min. Next, markers for endosomes, endoplasmic reticulum (ER), golgi apparatus (GA), lysosomes, and proteasomes and Fc receptors, microtubules, and actin were monitored by immunofluorescence. Labeled albumin co‐localization with endosomes was gradually and significantly increased and it was significantly higher than with the other markers at any timepoint. Albumin, placed on inside of the Transwell membrane, diffused through HRGEC monolayers during a 360 min incubation period. This transportation of albumin through HRGECs was inhibited by methyl beta cyclodextrin (MBCD), a caveolae disrupting agent. MBCD also decreased albuminuria, causing decreased caveolin‐1 (Cav‐1) expression on glomerular capillaries, in puromycin aminonucleoside induced nephrotic mice. Albumin transcytosis depends on early endosomes, but not on other organelles, Fc receptors, or cytoskeletal components. Caveolae disruption prevented albumin transportation through HRGECs and decreased albuminuria in nephrotic mice. This newly described caveolae‐dependent albumin pathway through glomerular endothelial cells is a potential pathogenetic mechanism for albuminuria, independent of the fenestrae. After endocytosis through caveolae into glomerular endothelial cells, albumin was transported to early endosome to sort to bypass lysosomes, proteasomes, the golgi apparatus, and endoplasmic reticulum, and is then transported to the other side of the cells. This newly described caveolae‐dependent albumin pathway through glomerular endothelial cells is a potential pathogenetic mechanism for albuminuria, independent of the fenestrae.

  May 15, 2017   doi: 10.1002/jcp.25817   open full text
• Individual‐specific variation in the respiratory activities of HMECs and their bioenergetic response to IGF1 and TNFα.
  Sallie S. Schneider, Elizabeth M. Henchey, Nazneen Sultana, Stephanie M. Morin, D. Joseph Jerry, Grace Makari‐Judson, Giovanna M. Crisi, Richard B. Arenas, Melissa Johnson, Holly S. Mason, Nagendra Yadava.
  Journal of Cellular Physiology. May 15, 2017
  Metabolic reprograming is a hallmark of cancer cells. However, the roles of pre‐existing differences in normal cells metabolism toward cancer risk is not known. In order to assess pre‐existing variations in normal cell metabolism, we have quantified the inter‐individual variation in oxidative metabolism of normal primary human mammary epithelial cells (HMECs). We then assessed their response to selected cytokines such as insulin growth factor 1 (IGF1) and tumor necrosis factor alpha (TNFα), which are associated with breast cancer risk. Specifically, we compared the oxidative metabolism of HMECs obtained from women with breast cancer and without cancer. Our data show considerable inter‐individual variation in respiratory activities of HMECs from different women. A bioenergetic parameter called pyruvate‐stimulated respiration (PySR) was identified as a key distinguishing feature of HMECs from women with breast cancer and without cancer. Samples showing PySR over 20% of basal respiration rate were considered PySR+ve and the rest as PySR−ve. By this criterion, HMECs from tumor‐affected breasts (AB) and non‐tumor affected breasts (NAB) of cancer patients were mostly PySR−ve (88% and 89%, respectively), while HMECs from non‐cancer patients were mostly PySR+ve (57%). This suggests that PySR−ve/+ve phenotypes are individual‐specific and are not caused by field effects due to the presence of tumor. The effects of IGF1 and TNFα treatments on HMECs revealed that both suppressed respiration and extracellular acidification. In addition, IGF1 altered PySR−ve/+ve phenotypes. These results reveal individual‐specific differences in pyruvate metabolism of normal breast epithelial cells and its association with breast cancer risk. We describe individual‐specific variation in respiratory activity and pyruvate metabolism of normal HMECs from tumor affected (AB) and non‐affected breasts (NAB). Our analyses identified a bioenergetic parameter, the pyruvate‐stimulated respiration (PySR), which distinguishes cells from cancer patients versus mammoplasty patients that did not have any cancer history.

  May 15, 2017   doi: 10.1002/jcp.25932   open full text
• Pathogenic Role of Associated Adherent‐Invasive Escherichia coli in Crohn's Disease.
  Giuseppe Mazzarella, Angelica Perna, Angela Marano, Angela Lucariello, Vera Rotondi Aufiero, Alida Sorrentino, Raffaele Melina, Germano Guerra, Fabio Silvio Taccone, Gaetano Iaquinto, Antonio De Luca.
  Journal of Cellular Physiology. May 15, 2017
  Several lines of evidence suggest that adherent‐invasive Escherichia coli (AIEC) strains play an important role in Crohn's disease (CD). The objective of this study was to investigate the pathogenic role of two AIEC strains, LF82 and O83:H1, in CD patients. Organ cultures of colonic biopsies from patients were set up to assess the effects of LF82 and O83:H1 on the expression of CEACAM6, LAMP1, HLA‐DR, ICAM1 by immunohistochemistry and of IL‐8, IFNʏ, and TNF‐α genes by RT‐PCR. Moreover, on Caco2 cells, we analyzed the cell cycle, the expression of MGMT and DNMT1 genes, and DNA damage induced by LF82 and O83:H1, by FACS, RT‐PCR, and DAPI staining, respectively. Epithelial and lamina propria mononuclear cells (LPMNC) expression of CEACAM6 and LAMP1 were higher in biopsies cultured in the presence of both O83:H1 and LF82 than in biopsies cultured with non‐pathogenic E. coli. Both AIEC strains induced increased expression of ICAM‐1 on blood vessels and HLA‐DR on LPMNC. We observed higher levels of TNF‐α, IFN‐γ, and IL‐8 transcripts in biopsies cultured with both AIEC strains than in those cultured with NP. Both LF82 and O83:H1, block the cell cycle into S phase, inducing DNA damage, and modulate the expression of DNMT1 and MGMT genes. Our data suggest that LF82 and 083:H1 strains of E. coli are able to increase in CD colonic biopsies the expression of all the pro‐inflammatory cytokines and all the mucosal immune markers investigated. J. Cell. Physiol. 232: 2860–2868, 2017. © 2016 Wiley Periodicals, Inc. Adherent‐invasive Escherichia coli (AIEC) strains play an important role in Crohn's disease (CD). The objective of this study was to investigate the pathogenic role of two AIEC strains, LF82 and O83:H1, in CD patients. LF82 and 083:H1 strains of E. coli are able to increase in CD colonic biopsies the expression of all the pro‐inflammatory cytokines and all the mucosal immune markers investigated.

  May 15, 2017   doi: 10.1002/jcp.25717   open full text
• Activating transcription factor 3 (ATF3) protects against lipopolysaccharide‐induced acute lung injury via inhibiting the expression of TL1A.
  Lanlan Qian, Yunfeng Zhao, Liang Guo, Shaoying Li, Xueling Wu.
  Journal of Cellular Physiology. May 15, 2017
  Excessive inflammatory responses are critical in the pathogenesis of acute lung injury (ALI). Activating transcription factor 3 (ATF3) is a stress‐induced transcriptional regulator that is a negative regulator of inflammatory responses. Therefore, we investigated the role and signaling pathways of ATF3 in lipopolysaccharide (LPS)‐induced ALI in mice. The mouse macrophage RAW264.7 cells were cultured on HTS 24‐Transwell filter plates in presence of ATF3 siRNA before exposure to LPS. ATF3 knock‐out (KO) and wild type (WT) mice were challenged by intra‐peritoneal injection of LPS (15 mg/kg). Gene analysis was used to analyze differential gene expression between ATF3 KO and WT mice. LPS increased the expression of ATF3 in RAW264.7 cells and in lung tissues of mice, The concentration of TNFα and IL‐6 was significantly increased in ATF3 siRNA‐treated RAW264.7 cells compared to control cells after LPS stimulation. The concentration of TNFα, IL‐6 and IL‐1β in serum and lung tissue of ATF3 KO mice was significantly increased compared to ATF3 WT mice. In addition, the lung wet/dry weight and BALF protein were significantly increased in ATF3 KO mice after LPS injection at 6, 24, and 48 hr. The survival of ATF3 KO mice significantly decreased. Differential gene analysis showed that TL1A was highly expressed in LPS‐induced lung tissues of ATF3 KO mice.Moreover, ATF3 down‐regulated the expression of TL1A in RAW264.7 cells and in lung tissues. These findings suggest that ATF3 protects against LPS‐induced ALI via inhibiting TL1A expression. Excessive inflammatory responses are critical in the pathogenesis of acute lung injury (ALI). Activating transcription factor 3 (ATF3) is a stress‐induced transcriptional regulator that is a negative regulator of inflammatory responses.The survival of ATF3 KO mice significantly decreased. Differential gene analysis showed that TL1A was highly expressed in LPS‐induced lung tissues of ATF3 KO mice.Moreover, ATF3 down‐regulated the expression of TL1A in RAW264.7 cells and in lung tissues.These findings suggest that ATF3 protects against LPS‐induced ALI via inhibiting TL1A expression.

  May 15, 2017   doi: 10.1002/jcp.25849   open full text
• MicroRNAs: Potential candidates for diagnosis and treatment of colorectal cancer.
  Abdullah Moridikia, Hamed Mirzaei, Amirhossein Sahebkar, Jafar Salimian.
  Journal of Cellular Physiology. May 15, 2017
  Colorectal cancer (CRC) is known as the third common cancer worldwide and an important public health problem in different populations. Several genetics and environmental risk factors are involved in the development and progression of CRC including chromosomal abnormalities, epigenetic alterations, and unhealthy lifestyle. Identification of risk factors and biomarkers could lead to a better understanding of molecular pathways involved in CRC pathogenesis. MicroRNAs (miRNAs) are important regulatory molecules which could affect a variety of cellular and molecular targets in CRC. A large number of studies have indicated deregulations of some known tissue‐specific miRNAs, for example, miR‐21, miR‐9, miR‐155, miR‐17, miR‐19, let‐7, and miR‐24 as well as circulating miRNAs, for example, miR‐181b, miR‐21, miR‐183, let‐7g, miR‐17, and miR‐126, in patients with CRC. In the current review, we focus on the findings of preclinical and clinical studies performed on tissue‐specific and circulating miRNAs as diagnostic biomarkers and therapeutic targets for the detection of patients at various stages of CRC.

  May 15, 2017   doi: 10.1002/jcp.25801   open full text
• NMDA receptor subunit composition controls dendritogenesis of hippocampal neurons through CAMKII, CREB‐P, and H3K27ac.
  Fernando J. Bustos, Nur Jury, Pablo Martinez, Estibaliz Ampuero, Matias Campos, Sebastian Abarzúa, Karen Jaramillo, Susanne Ibing, Muriel D. Mardones, Henny Haensgen, Julia Kzhyshkowska, Maria Florencia Tevy, Rachael Neve, Magdalena Sanhueza, Lorena Varela‐Nallar, Martín Montecino, Brigitte van Zundert.
  Journal of Cellular Physiology. May 11, 2017
  Dendrite arbor growth, or dendritogenesis, is choreographed by a diverse set of cues, including the NMDA receptor (NMDAR) subunits NR2A and NR2B. While NR1NR2B receptors are predominantly expressed in immature neurons and promote plasticity, NR1NR2A receptors are mainly expressed in mature neurons and induce circuit stability. How the different subunits regulate these processes is unclear, but this is likely related to the presence of their distinct C‐terminal sequences that couple different signaling proteins. Calcium‐calmodulin‐dependent protein kinase II (CaMKII) is an interesting candidate as this protein can be activated by calcium influx through NMDARs. CaMKII triggers a series of biochemical signaling cascades, involving the phosphorylation of diverse targets. Among them, the activation of cAMP response element‐binding protein (CREB‐P) pathway triggers a plasticity‐specific transcriptional program through unknown epigenetic mechanisms. Here, we found that dendritogenesis in hippocampal neurons is impaired by several well‐characterized constructs (i.e., NR2B‐RS/QD) and peptides (i.e., tatCN21) that specifically interfere with the recruitment and interaction of CaMKII with the NR2B C‐terminal domain. Interestingly, we found that transduction of NR2AΔIN, a mutant NR2A construct with increased interaction to CaMKII, reactivates dendritogenesis in mature hippocampal neurons in vitro and in vivo. To gain insights into the signaling and epigenetic mechanisms underlying NMDAR‐mediated dendritogenesis, we used immunofluorescence staining to detect CREB‐P and acetylated lysine 27 of histone H3 (H3K27ac), an activation‐associated histone tail mark. In contrast to control mature neurons, our data shows that activation of the NMDAR/CaMKII/ERK‐P/CREB‐P signaling axis in neurons expressing NR2AΔIN is not correlated with increased nuclear H3K27ac levels. Dendrite arbor growth, or dendritogenesis, is choreographed by a diverse set of cues, including the NMDA receptor (NMDAR) subunits NR2A and NR2B. Here we found that dendritogenesis in hippocampal neurons is controlled by the interaction between NMDAR subunits and CaMKII in a developmental dependent manner.

  May 11, 2017   doi: 10.1002/jcp.25843   open full text
• Estrogens and androgens inhibit association of RANKL with the pre‐osteoblast membrane through post‐translational mechanisms.
  Anthony Martin, Jiali Yu, Jian Xiong, Aysha B. Khalid, Benita Katzenellenbogen, Sung Hoon Kim, John A. Katzenellenbogen, Suchinda Malaivijitnond, Yankel Gabet, Susan A. Krum, Baruch Frenkel.
  Journal of Cellular Physiology. May 11, 2017
  We have recently demonstrated that RUNX2 promoted, and 17β‐Estradiol (E2) diminished, association of RANKL with the cell membrane in pre‐osteoblast cultures. Here we show that, similar to E2, dihydrotestosterone (DHT) diminishes association of RANKL, and transiently transfected GFP‐RANKL with the pre‐osteoblast membrane without decreasing total RANKL mRNA or protein levels. Diminution of membrane‐associated RANKL was accompanied with marked suppression of osteoclast differentiation from co‐cultured pre‐osteoclasts, even though DHT increased, not decreased, RANKL concentrations in pre‐osteoblast conditioned media. A marked decrease in membrane‐associated RANKL was observed after 30 min of either E2 or DHT treatment, and near‐complete inhibition was observed by 1 hr, suggesting that the diminution of RANKL membrane association was mediated through non‐genomic mechanisms. Further indicating dispensability of nuclear action of estrogen receptor, E2‐mediated inhibition of RANKL membrane association was mimicked by an estrogen dendrimer conjugate (EDC) that cannot enter the cell nucleus. Finally, the inhibitory effect of E2 and DHT on RANKL membrane association was counteracted by the MMP inhibitor NNGH, and the effect of E2 (and not DHT) was antagonized by the Src inhibitor SU6656. Taken together, these results suggest that estrogens and androgens inhibit osteoblast‐driven osteoclastogenesis through non‐genomic mechanism(s) that entail, MMP‐mediated RANKL dissociation from the cell membrane. Association of RANKL with the pre‐osteoblast membrane is dramatically reduced within minutes of treatment with either estradiol or dihydrotestosterone. The effect of estradiol is MMP‐ and Src‐dependent, suggesting RANKL ectodomain shedding, and is mimicked by an estrogen dendrimer conjugate that cannot enter the cell nucleus.

  May 11, 2017   doi: 10.1002/jcp.25862   open full text
• MicroRNA‐224‐5p regulates adipocyte apoptosis induced by TNFα via controlling NF‐κB activation.
  Renli Qi, Jinxiu Huang, Qi Wang, Hong Liu, Ruisheng Wang, Jing Wang, Feiyun Yang.
  Journal of Cellular Physiology. May 10, 2017
  Tumor necrosis factor (TNF) α can induce cell apoptosis and activate nuclear transcription (NF)‐κB in different cell types. Activated NF‐κB further promotes or suppresses cellular apoptosis in different cases. The present study explored the effect of activated NF‐κB on adipocyte apoptosis induced by TNFα and which microRNAs (miRNAs) were involved in the process. Our findings demonstrated that treatment of differentiated 3T3‐L1 adipocytes with TNFα (20ng/mL) rapidly activated NF‐κB and induced moderate apoptosis. Pyrrolidinedithiocarbamic acid (PDTC, 60µM), a specific NF‐κB inhibitor, abated NF‐κB activation that rendered the adipocytes vulnerable to TNFα‐induced apoptosis. Dozens of miRNAs exhibited significant expression changes following TNFα treatment and the addition of PDTC. In which, miRNA‐224‐5p (miR‐224) was up‐regulated by TNFα exposure but down‐regulated by PDTC addition. Furthermore, over‐expression of miR‐224 promoted NF‐κB activation and prevented the adipocyte apoptosis induced by TNFα, while miR‐224 deficiency showed the opposite effects. The TRAF‐associated NF‐κB activator (TANK) gene was identified as a direct target of miR‐224 by computational and luciferase reporter assays. Additionally, silencing the TANK gene by the small interfering RNA similarly promoted NF‐κB activation and attenuated the cellular apoptosis. In conclusion, these findings demonstrate that miR‐224 plays an essential role in adipocyte apoptosis caused by TNFα through control of NF‐κB activation via targeting the TANK gene. This article is protected by copyright. All rights reserved

  May 10, 2017   doi: 10.1002/jcp.25992   open full text
• Leucine elicits myotube hypertrophy and enhances maximal contractile force in tissue engineered skeletal muscle in vitro.
  Neil R.W. Martin, Mark C. Turner, Robert Farrington, Darren J. Player, Mark P. Lewis.
  Journal of Cellular Physiology. May 08, 2017
  The amino acid leucine is thought to be important for skeletal muscle growth by virtue of its ability to acutely activate mTORC1 and enhance muscle protein synthesis, yet little data exist regarding its impact on skeletal muscle size and its ability to produce force. We utilized a tissue engineering approach in order to test whether supplementing culture medium with leucine could enhance mTORC1 signaling, myotube growth, and muscle function. Phosphorylation of the mTORC1 target proteins 4EBP‐1 and rpS6 and myotube hypertrophy appeared to occur in a dose dependent manner, with 5 and 20 mM of leucine inducing similar effects, which were greater than those seen with 1 mM. Maximal contractile force was also elevated with leucine supplementation; however, although this did not appear to be enhanced with increasing leucine doses, this effect was completely ablated by co‐incubation with the mTOR inhibitor rapamycin, showing that the augmented force production in the presence of leucine was mTOR sensitive. Finally, by using electrical stimulation to induce chronic (24 hr) contraction of engineered skeletal muscle constructs, we were able to show that the effects of leucine and muscle contraction are additive, since the two stimuli had cumulative effects on maximal contractile force production. These results extend our current knowledge of the efficacy of leucine as an anabolic nutritional aid showing for the first time that leucine supplementation may augment skeletal muscle functional capacity, and furthermore validates the use of engineered skeletal muscle for highly‐controlled investigations into nutritional regulation of muscle physiology. The effect of the amino acid leucine on myotube size and muscle force production were tested in engineered skeletal muscle in vitro. Leucine activated mTORC1 signaling and augmented contractile force, effects which were completely blunted by co‐incubation with rapamycin. Chronic contractions (24 hr) alongside leucine treatment further increased muscle force, but not myotube size. This data provides further evidence of the efficacy of leucine in regulating muscle size and function, and may prove a useful strategy to improve the therapeutic potential of engineered skeletal muscle.

  May 08, 2017   doi: 10.1002/jcp.25960   open full text
• E2F1interactive with BRCA1 pathway induces HCC two different small molecule metabolism or cell cycle regulation via mitochondrion or CD4+T to cytosol.
  Qingchun Chen, Lin Wang, Minghu Jiang, Juxiang Huang, Zhenfu Jiang, Haitao Feng, Zhili Ji.
  Journal of Cellular Physiology. May 05, 2017
  Breast cancer 1 (BRCA1) and E2F transcription factor 1 (E2F1) are related to metabolism and cell cycle regulation. However, the corresponding mechanism is not clear in HCC. High BRCA1 direct pathway was constructed with 11 molecules from E2F1 feedback‐interactive network in HCC by GRNInfer based on 39 Pearson mutual positive corelation CC >0.25 molecules with E2F1. Integration of GRNInfer with GO, KEGG, BioCarta, GNF_U133A, UNIGENE_EST, Disease, GenMAPP databases by DAVID and MAS 3.0, E2F1 feedback‐interactive BRCA1 indirect mitochondrion to cytosol pathway was identified as upstream LAPTM4B activation, feedback UNG, downstream BCAT1‐HIST1H2AD‐TK1 reflecting protein and DNA binding with enrichment of small molecule metabolism; The corresponding BRCA1 indirect membrane to cytosol pathway as upstream CCNB2‐NUSAP1 activation, feedback TTK‐HIST1H2BJ‐CENPF, downstream MCM4‐TK1 reflecting ATP and microtubule binding with enrichment of CD4+T‐related cell cycle regulation in HCC. Therefore, we propose that E2F1 interactive with BRCA1 pathway induces HCC two different small molecule metabolism or cell cycle regulation via mitochondrion or CD4+T to cytosol. Knowledge analysis demonstrates our E2F1 feedback‐interactive BRCA1 pathway wide disease distribution and reflects a novel common one of tumor and cancer. This article is protected by copyright. All rights reserved

  May 05, 2017   doi: 10.1002/jcp.25988   open full text
• Direct Delivery of Recombinant Pin1 Protein Rescued Osteoblast Differentiation of Pin1‐Deficient Cells.
  Woo‐Jin Kim, Rabia Islam, Bong‐Soo Kim, Young‐Dan Cho, Won‐Joon Yoon, Jeong‐Hwa Baek, Kyung‐Mi Woo, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. May 05, 2017
  Pin1 is a peptidyl prolyl cis‐trans isomerase that specifically binds to the phosphoserine‐proline or phosphothreonine‐proline motifs of several proteins. We reported that Pin1 plays a critical role in the fate determination of Smad1/5, Runx2, and β‐catenin that are indispensable nuclear proteins for osteoblast differentiation. Though several chemical inhibitors has been discovered for Pin1, no activator has been reported as of yet. In this study, we directly introduced recombinant Pin1 protein successfully into the cytoplasm via fibroin nanoparticle encapsulated in cationic lipid. This nanoparticle‐lipid complex delivered its cargo with a high efficiency and a low cytotoxicity. Direct delivery of Pin1 leads to increased Runx2 and Smad signaling and resulted in recovery of the osteogenic marker genes expression and the deposition of mineral in Pin1‐deficient cells. These result indicated that a direct Pin1 protein delivery method could be a potential therapeutics for the osteopenic diseases. J. Cell. Physiol. 232: 2798–2805, 2017. © 2016 Wiley Periodicals, Inc. Direct delivery of Pin1 leads to increased Runx2 and Smad signaling and resulted in recovery of the osteogenic marker genes expression and the deposition of mineral in Pin1‐deficient cells.

  May 05, 2017   doi: 10.1002/jcp.25673   open full text
• The diversified function and potential therapy of ectopic olfactory receptors in non‐olfactory tissues.
  Zhe Chen, Hong Zhao, Nian Fu, Linxi Chen.
  Journal of Cellular Physiology. May 05, 2017
  Olfactory receptors (ORs) are mainly distributed in olfactory neurons and play a key role in detecting volatile odorants, eventually resulting in the production of smell perception. Recently, it is also reported that ORs are expressed in non‐olfactory tissues including heart, lung, sperm, skin, and cancerous tissues. Interestingly, ectopic ORs are associated with the development of diseases in non‐olfactory tissues. For instance, ectopic ORs initiate the hypoxic ventilatory responses and maintain the oxygen homeostasis of breathing in the carotid body when oxygen levels decline. Ectopic ORs induce glucose homeostasis in diabetes. Ectopic ORs regulate systemic blood pressure by increasing renin secretion and vasodilation. Ectopic ORs participate in the process of tumor cell proliferation, apoptosis, metastasis, and invasiveness. Ectopic ORs accelerate the occurrence of obesity, angiogenesis and wound‐healing processes. Ectopic ORs affect fetal hemoglobin levels in sickle cell anemia and thalassemia. Finally, we also elaborate some ligands targeting for ORs. Obviously, the diversified function and related signal pathway of ectopic ORs may play a potential therapeutic target in non‐olfactory tissues. Thus, this review focuses on the latest research results about the diversified function and therapeutic potential of ectopic ORs in non‐olfactory tissues. This review focuses on the latest research results about the diversified function and therapeutic potential of ectopic ORs in non‐olfactory tissues.

  May 05, 2017   doi: 10.1002/jcp.25929   open full text
• Irisin plays a pivotal role to protect the heart against ischemia and reperfusion injury.
  Hao Wang, Yu Tina Zhao, Shouyan Zhang, Patrycja M Dubielecka, Jianfeng Du, Naohiro Yano, Y. Eugene Chin, Shougang Zhuang, Gangjian Qin, Ting C Zhao.
  Journal of Cellular Physiology. May 03, 2017
  Irisin, a newly identified hormone, is critical to modulating body metabolism, thermogenesis and reducing oxidative stresses. However, whether irisin protects the heart against myocardial ischemia and reperfusion (I/R) injury remains unknown. In this study, we determine the effect of irisin on myocardial I/R injury in the Langendorff perfused heart and cultured myocytes. Adult C57/BL6 mice were treated with irisin (100 mg/kg) or vehicle for 30 min to elicit preconditioning. The isolated hearts were subjected to 30 min ischemia followed by 30 min reperfusion. Left ventricular function was measured and infarction size were determined using by tetrazolium staining. Western blot was employed to determine myocardial SOD‐1, active‐caspase 3, annexin V, p38, and phospho‐p38. H9c2 cardiomyoblasts were exposed to hypoxia and reoxygenation for assessment of the effects of irisin on mitochondrial respiration and mitochondrial permeability transition pore (mPTP). Irisin treatment produced remarkable improvements in ventricular functional recovery, as evident by the increase in RPP and attenuation in LVEDP. As compared to the vehicle treatment, irisin resulted in a marked reduction of myocardial infarct size. Notably, irisin treatment increased SOD‐1 and p38 phosphorylation, but suppressed levels of active‐caspase 3, cleaved PARP, and annexin V. In cardiomyoblasts exposed to hypoxia/reoxygenation, irisin treatment significantly attenuated hypoxia/reoxygenation (H/R), as indicated by the reduction of lactate dehydrogenase (LDH) leakage and apoptotic cardiomyocytes. Furthermore, irisin treatments suppressed the opening of mPTP, mitochondrial swelling, and protected mitochondria function. Our results indicate that irisin serves as a novel approach to eliciting cardioprotection, which is associated with the improvement of mitochondrial function. Irisin treatment produced remarkable improvements in ventricular functional recovery and resulted in a marked reduction of myocardial infarct size. Furthermore, irisin treatments suppressed the opening of mPTP, mitochondrial swelling, and protected mitochondria function. Our results indicate that irisin serves as a novel approach to eliciting cardioprotection, which is associated with the improvement of mitochondrial function.

  May 03, 2017   doi: 10.1002/jcp.25857   open full text
• MicroRNA‐195‐5p Regulates Osteogenic Differentiation of Periodontal Ligament Cells Under Mechanical Loading.
  Maolin Chang, Heng Lin, Haidi Fu, Beike Wang, Guangli Han, Mingwen Fan.
  Journal of Cellular Physiology. May 03, 2017
  Osteogenic differentiation and bone formation are tightly regulated by several factors, including microRNAs (miRNAs). However, miRNA expression patterns and function during mechanical loading‐induced osteogenic differentiation of human periodontal ligament cells (PDLCs) remain unclear. Here, we investigated the differential expression of miRNA‐195‐5p in the periodontal tissues of mice under orthodontic mechanical loading and in primary human PDLCs exposed to a simulated tension strain. The miR‐195‐5p was observed to be down‐regulated and negatively correlated with osteogenic differentiation. Overexpression of miR‐195‐5p significantly inhibited PDLC differentiation under cyclic tension strain (CTS), whereas the functional inhibition of miR‐195‐5p yielded an opposite effect. Further experiments confirmed that WNT family member 3A (WNT3A), fibroblast growth factor 2 (FGF2), and bone morphogenetic protein receptor‐1A (BMPR1A), proteins important for osteogenic activity and stability, were direct targets of miR‐195‐5p. Mechanical loading increased the WNT3A, FGF2, and BMPR1A protein levels, while miR‐195‐5p inhibited WNT3A, FGF2, and BMPR1A protein expression. WNT, FGF, and BMP signaling were involved in osteogenic differentiation of PDLCs under CTS. Further study confirmed that reintroduction of WNT3A and BMPR1A can rescue the inhibition of miR‐195‐5p on osteogenic differentiation of PDLCs. Our findings are the first to demonstrate that miR‐195‐5p is a mechanosensitive gene that plays an important role in mechanical loading‐induced osteogenic differentiation and bone formation. During orthodontic treatment, periodontal ligament cells can differentiate into osteoblasts under tension strain. The miR‐195‐5p was down‐regulated under tension strain, leading to the disinhibition of WNT3A, FGF2, and BMPR1A, which associated with othodonitc force induced bone formation.

  May 03, 2017   doi: 10.1002/jcp.25856   open full text
• Deubiquitinating enzyme USP22 positively regulates c‐Myc stability and tumorigenic activity in mammalian and breast cancer cells.
  Dongyeon Kim, Ahyoung Hong, Hye In Park, Woo Hyun Shin, Lang Yoo, Seo Jeong Jeon, Kwang Chul Chung.
  Journal of Cellular Physiology. May 03, 2017
  The proto‐oncogene c‐Myc has a pivotal function in growth control, differentiation, and apoptosis and is frequently affected in human cancer, including breast cancer. Ubiquitin‐specific protease 22 (USP22), a member of the USP family of deubiquitinating enzymes (DUBs), mediates deubiquitination of target proteins, including histone H2B and H2A, telomeric repeat binding factor 1, and cyclin B1. USP22 is also a component of the mammalian SAGA transcriptional co‐activating complex. In this study, we explored the functional role of USP22 in modulating c‐Myc stability and its physiological relevance in breast cancer progression. We found that USP22 promotes deubiquitination of c‐Myc in several breast cancer cell lines, resulting in increased levels of c‐Myc. Consistent with this, USP22 knockdown reduces c‐Myc levels. Furthermore, overexpression of USP22 stimulates breast cancer cell growth and colony formation, and increases c‐Myc tumorigenic activity. In conclusion, the present study reveals that USP22 in breast cancer cell lines increases c‐Myc stability through c‐Myc deubiquitination, which is closely correlated with breast cancer progression. The present study reveals that deubiquitinating enzyme USP22 in mammalia and breast cancer cell lines increases c‐Myc stability through c‐Myc deubiquitination, which is closely correlated with breast cancer progression.

  May 03, 2017   doi: 10.1002/jcp.25841   open full text
• An advanced tri‐culture model to evaluate the dynamic interplay among osteoblasts, osteoclasts, and endothelial cells.
  Stefania Pagani, Paola Torricelli, Francesca Veronesi, Francesca Salamanna, Simona Cepollaro, Milena Fini.
  Journal of Cellular Physiology. May 03, 2017
  The dynamic metabolism and the numerous roles of bone tissue necessitate a suitable in vitro model to represent them. In order to investigate the interaction among the several cell types composing bone microenvironment, we studied a tri‐culture model including human osteoblasts (OBs), osteoclasts (OCs), and endothelial cells (HUVEC). While OBs are essential for bone deposition and OCs for bone resorption, the vasculature is necessary to provide growth factors, nutrients, and oxygen in the mature tissue. The results of this study showed a strong mutual influence between OBs, OCs, and HUVEC in term of proliferation, viability, and activity (release of ALP, Coll I, OPG, RANKL, VEGF, CTSK, TGFβ, and IL‐6). The behavior of the single cultures demonstrated to be different compared to the bi‐ or tri‐cultures and depending on the cell types involved: the coexistence of OBs and OCs stimulated the synthetic activity of both cell types, while the presence of HUVEC induced a stimulating role for OBs but mainly an inhibitory effect for OC. In addition, evidence of the effects of OBs and OCs on HUVEC is highlighted by their morphology: regular and able to “sketch” little vessels in presence of OBs, more disorganized and heterogeneous in presence of OCs. Taken together, these observations well characterize an advanced cellular model to be used as starting point for mimicking bone microenvironment in vivo, thus reducing the use of animals in the preclinical phase and offering a more reliable tool to test new and innovative biomaterials. In vitro model of direct co‐colture showing the mutual influence among osteoblast for bone deposition, osteoclast for bone resorption and endothelial cells for angiogenesis, as starting point in mimicking in vivo microenvironment.

  May 03, 2017   doi: 10.1002/jcp.25875   open full text
• Blocking fibrotic signaling in fibroblasts from patients with carpal tunnel syndrome.
  Yoshiaki Yamanaka, Anne Gingery, Gosuke Oki, Tai‐Hua Yang, Chunfeng Zhao, Peter C. Amadio.
  Journal of Cellular Physiology. May 03, 2017
  Fibrosis of the subsynovial connective tissue (SSCT) in carpal tunnel syndrome (CTS) patients is increasingly recognized as an important aspect of CTS pathophysiology. In this study, we evaluated the effect of blocking profibrotic pathways in fibroblasts from the SSCT in CTS patients. Fibroblasts were stimulated with transforming growth factor β1 (TGF‐β1), and then treated either with a specific fibrosis pathway inhibitor targeting TGF‐β receptor type 1 (TβRI), platelet‐derived growth factor receptor (PDGFR), epidermal growth factor receptor (EGFR), or vascular endothelial growth factor receptor (VEGFR). Fibrosis array and quantitative real‐time polymerase chain reaction of fibrotic genes were evaluated. Array gene expression analysis revealed significant down‐regulation of multiple fibrotic genes after treatment with TβRI, PDGFR, and VEGFR inhibitors. No array fibrotic genes were significantly down‐regulated with EGFR inhibition. Further gene expression analysis of known CTS fibrosis markers collagen type I A2 (Col1), collagen type III A1 (Col3), connective tissue growth factor (CTGF), and SERPINE1 showed significantly down‐regulation after TβRI inhibition. In contrast, VEGFR inhibition significantly down‐regulated CTGF and SERPINE1, whereas, PDGFR and EGFR inhibition significantly down‐regulated Col3. Taken together the inhibition of TβRI appears to be the primary mediator of fibrotic gene expression in fibroblasts from CTS patients. TGF‐β/Smad activity was further evaluated, and as expected inhibition of Smad activity was significantly down‐regulated after inhibition of TβRI, but not with PDGFR, VEGFR, or EGFR inhibition. These results indicate that local therapies specifically targeting TGF‐β signaling alone or in combination offer the potential of a novel local antifibrosis therapy for patients with CTS. In this study, we evaluated the effect of blocking profibrotic pathways by receptor blocking agents in fibroblasts from patients with carpal tunnel syndrome (CTS). There was significant down‐regulation of multiple fibrotic genes after treatment with a TFFβ1R (SD208) inhibitor, with smaller effects from PDGFR (AG1296) and VEGFR (Axitinib) inhibitors. These results indicate that local therapies specifically targeting TGF‐β signaling, alone or in combination with other pathways, offer the potential of a novel local anti‐fibrosis therapy for patients with CTS.

  May 03, 2017   doi: 10.1002/jcp.25901   open full text
• Rapamycin attenuates BAFF‐extended proliferation and survival via disruption of mTORC1/2 signaling in normal and neoplastic B‐lymphoid cells.
  Qingyu Zeng, Shanshan Qin, Hai Zhang, Beibei Liu, Jiamin Qin, Xiaoxue Wang, Ruijie Zhang, Chunxiao Liu, Xiaoqing Dong, Shuangquan Zhang, Shile Huang, Long Chen.
  Journal of Cellular Physiology. May 03, 2017
  B cell activating factor from the TNF family (BAFF) stimulates B‐cell proliferation and survival, but excessive BAFF promotes the development of aggressive B cells leading to malignant and autoimmune diseases. Recently, we have reported that rapamycin, a macrocyclic lactone, attenuates human soluble BAFF (hsBAFF)‐stimulated B‐cell proliferation/survival by suppressing mTOR‐mediated PP2A‐Erk1/2 signaling pathway. Here, we show that the inhibitory effect of rapamycin on hsBAFF‐promoted B cell proliferation/survival is also related to blocking hsBAFF‐stimulated phosphorylation of Akt, S6K1, and 4E‐BP1, as well as expression of survivin in normal and B‐lymphoid (Raji and Daudi) cells. It appeared that both mTORC1 and mTORC2 were involved in the inhibitory activity of rapamycin, as silencing raptor or rictor enhanced rapamycin's suppression of hsBAFF‐induced survivin expression and proliferation/viability in B cells. Also, PP242, an mTORC1/2 kinase inhibitor, repressed survivin expression, and cell proliferation/viability more potently than rapamycin (mTORC1 inhibitor) in B cells in response to hsBAFF. Of interest, ectopic expression of constitutively active Akt (myr‐Akt) or constitutively active S6K1 (S6K1‐ca), or downregulation of 4E‐BP1 conferred resistance to rapamycin's attenuation of hsBAFF‐induced survivin expression and B‐cell proliferation/viability, whereas overexpression of dominant negative Akt (dn‐Akt) or constitutively hypophosphorylated 4E‐BP1 (4EBP1‐5A), or downregulation of S6K1, or co‐treatment with Akt inhibitor potentiated the inhibitory effects of rapamycin. The findings indicate that rapamycin attenuates excessive hsBAFF‐induced cell proliferation/survival via blocking mTORC1/2 signaling in normal and neoplastic B‐lymphoid cells. Our data underscore that rapamycin may be a potential agent for preventing excessive BAFF‐evoked aggressive B‐cell malignancies and autoimmune diseases. Rapamycin, a macrocyclic lactone, prevented B cells from hsBAFF‐promoted proliferation/survival, not only by targeting mTORC1‐mediated S6K1/4E‐BP1 pathways, but also via targeting mTORC2‐mediated Akt pathway. This study highlights that rapamycin may be a potential agent for preventing excessive BAFF‐evoked aggressive B‐cell malignancies and autoimmune diseases.

  May 03, 2017   doi: 10.1002/jcp.25913   open full text
• HDAC1 and HDAC3 underlie dynamic H3K9 acetylation during embryonic neurogenesis and in schizophrenia‐like animals.
  Josef Večeřa, Eva Bártová, Jana Krejčí, Soňa Legartová, Denisa Komůrková, Jana Rudá‐Kučerová, Tibor Štark, Eva Dražanová, Tomáš Kašpárek, Alexandra Šulcová, Frank J. Dekker, Wiktor Szymanski, Christian Seiser, Georg Weitzer, Raphael Mechoulam, Vincenzo Micale, Stanislav Kozubek.
  Journal of Cellular Physiology. May 03, 2017
  Although histone acetylation is one of the most widely studied epigenetic modifications, there is still a lack of information regarding how the acetylome is regulated during brain development and pathophysiological processes. We demonstrate that the embryonic brain (E15) is characterized by an increase in H3K9 acetylation as well as decreases in the levels of HDAC1 and HDAC3. Moreover, experimental induction of H3K9 hyperacetylation led to the overexpression of NCAM in the embryonic cortex and depletion of Sox2 in the subventricular ependyma, which mimicked the differentiation processes. Inducing differentiation in HDAC1‐deficient mouse ESCs resulted in early H3K9 deacetylation, Sox2 downregulation, and enhanced astrogliogenesis, whereas neuro‐differentiation was almost suppressed. Neuro‐differentiation of (wt) ESCs was characterized by H3K9 hyperacetylation that was associated with HDAC1 and HDAC3 depletion. Conversely, the hippocampi of schizophrenia‐like animals showed H3K9 deacetylation that was regulated by an increase in both HDAC1 and HDAC3. The hippocampi of schizophrenia‐like brains that were treated with the cannabinoid receptor‐1 inverse antagonist AM251 expressed H3K9ac at the level observed in normal brains. Together, the results indicate that co‐regulation of H3K9ac by HDAC1 and HDAC3 is important to both embryonic brain development and neuro‐differentiation as well as the pathophysiology of a schizophrenia‐like phenotype. We showed that co‐regulation of H3K9 acetylation by histone deacetylases HDAC1 and HDAC3 is important to both embryonic brain development and neuro‐differentiation as well as the pathophysiology of a schizophrenia‐like phenotype. This knowledge contributes to a basic understanding of epigenetics of the brain.

  May 03, 2017   doi: 10.1002/jcp.25914   open full text
• Chrysin induces death of prostate cancer cells by inducing ROS and ER stress.
  Soomin Ryu, Whasun Lim, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. May 03, 2017
  Chrysin is a natural flavone found in numerous plant extracts, honey, and propolis that has multiple biological activities including anti‐cancer effects. Understanding of biological mechanisms mediated in response to chrysin in cancerous cells may provide novel insight into chemotherapeutic approaches with reduced side effects in cancers. In the present study, we investigated functional roles of chrysin in progression of prostate cancer cells using DU145 and PC‐3 cell lines. The results showed that chrysin induced apoptosis of cells evidenced by DNA fragmentation and increasing the population of both DU145 and PC‐3 cells in the sub‐G1 phase of the cell cycle. In addition, chrysin reduced expression of proliferating cell nuclear antigen in the prostate cancer cell lines compared to untreated prostate cancer cells. Moreover, chrysin induced loss of mitochondria membrane potential (MMP), while increasing production of reactive oxygen species (ROS) and lipid peroxidation in a dose‐dependent manner. Also, it induced endoplasmic reticulum (ER) stress through activation of unfolded protein response (UPR) proteins including PRKR‐like ER kinase (PERK), eukaryotic translation initiation factor 2α (eIF2α), and 78 kDa glucose‐regulated protein (GRP78) in DU145 and PC‐3 cells. The chrysin‐mediated intracellular signaling pathways suppressed phosphoinositide 3‐kinase (PI3K) and the abundance of AKT, P70S6K, S6, and P90RSK proteins, but stimulated mitogen‐activated protein kinases (MAPK) and activation of ERK1/2 and P38 proteins in the prostate cancer cells. Collectively, these results indicate that chrysin initiates cell death through induction of mitochondrial‐mediated apoptosis and ER stress, and regulation of signaling pathways responsible for proliferation of prostate cancer cells. Chrysin initiates cell death through induction of ROS production and ER stress, and regulation of MAPK signaling pathways responsible for proliferation of prostate cancer cells.

  May 03, 2017   doi: 10.1002/jcp.25861   open full text
• Transforming growth factor β induces bone marrow mesenchymal stem cell migration via noncanonical signals and N‐cadherin.
  Maria Jose Dubon, Jinyeong Yu, Sanghyuk Choi, Ki‐Sook Park.
  Journal of Cellular Physiology. May 03, 2017
  Transforming growth factor‐beta (TGF‐β) induces the migration and mobilization of bone marrow‐derived mesenchymal stem cells (BM‐MSCs) to maintain bone homeostasis during bone remodeling and facilitate the repair of peripheral tissues. Although many studies have reported the mechanisms through which TGF‐β mediates the migration of various types of cells, including cancer cells, the intrinsic cellular mechanisms underlying cellular migration, and mobilization of BM‐MSCs mediated by TGF‐β are unclear. In this study, we showed that TGF‐β activated noncanonical signaling molecules, such as Akt, extracellular signal‐regulated kinase 1/2 (ERK1/2), focal adhesion kinase (FAK), and p38, via TGF‐β type I receptor in human BM‐MSCs and murine BM‐MSC‐like ST2 cells. Inhibition of Rac1 by NSC23766 and Src by PP2 resulted in impaired TGF‐β‐mediated migration. These results suggested that the Smad‐independent, noncanonical signals activated by TGF‐β were necessary for migration. We also showed that N‐cadherin‐dependent intercellular interactions were required for TGF‐β‐mediated migration using functional inhibition of N‐cadherin with EDTA treatment and a neutralizing antibody (GC‐4 antibody) or siRNA‐mediated knockdown of N‐cadherin. However, N‐cadherin knockdown did not affect the global activation of noncanonical signals in response to TGF‐β. Therefore, these results suggested that the migration of BM‐MSCs in response to TGF‐β was mediated through N‐cadherin and noncanonical TGF‐β signals. N‐cadherin‐dependent intercellular interaction is required for TGF‐β‐mediated migration of both BM‐MSCs and the BM‐MSC like cell line, ST2. It is likely that in addition to N‐cadherin, non‐canonical TGF‐β signals including Akt, ERKs, p38, FAK, and Rac1 are also required for the cell migration.

  May 03, 2017   doi: 10.1002/jcp.25863   open full text
• Mitochondria‐targeted esculetin inhibits PAI‐1 levels by modulating STAT3 activation and miR‐19b via SIRT3: Role in acute coronary artery syndrome.
  Sujana Katta, Santosh Karnewar, Devayani Panuganti, Mahesh Kumar Jerald, B. K. S. Sastry, Srigiridhar Kotamraju.
  Journal of Cellular Physiology. May 03, 2017
  In this study we explored the microRNAs responsible for the regulation of PAI‐1 during LPS‐stimulated inflammation in human aortic endothelial cells and subsequently studied the effect of a newly synthesized mitochondria‐targeted esculetin (Mito‐Esc) that was shown for its anti‐atherosclerotic potential, in modulating PAI‐1 levels and its targeted miRs during angiotensin‐II‐induced atherosclerosis in ApoE−/− mice. LPS‐stimulated PAI‐1 was accompanied with an upregulation of miR‐19b and down‐regulation of miR‐30c. These effects of LPS on PAI‐1 were reversed in the presence of both parent esculetin and Mito‐Esc. However, the effect of Mito‐Esc was more pronounced in the regulation of PAI‐1. In addition, LPS‐stimulated PAI‐1 expression was significantly decreased in cells treated with Anti‐miR‐19b, thereby suggesting that miR‐19b co‐expression plays a key role in PAI‐1 regulation. The results also show that incubation of cells with Stattic, an inhibitor of STAT‐3, inhibited LPS‐stimulated PAI‐1 expression. Interestingly, knockdown of SIRT3, a mitochondrial biogenetic marker, enhanced PAI‐1 levels via modulation of miR‐19b and ‐30c. Mito‐Esc treatment significantly inhibited Ang‐II‐induced PAI‐1, possibly via altering miR‐19b and 30c in ApoE−/− mice. The association between PAI‐1, miR‐19b and ‐30c were further confirmed in plasma and microparticles isolated from patients suffering from acute coronary syndrome of various degrees. Taken together, LPS‐induced PAI‐1 involves co‐expression of miR‐19b and down regulation of miR‐30c, and Mito‐Esc treatment by modulating miR‐19b and miR‐30c through SIRT3 activation, inhibits PAI‐1 levels that, in part, contribute to its anti‐atherosclerotic effects. Moreover, there exists a strong positive correlation between miR‐19b and PAI‐1 in patients suffering from ST‐elevated myocardial infarction. LPS‐induced PAI‐1 involves co‐expression of miR‐19b and down regulation of miR‐30c. Mito‐Esc treatment by modulating miR‐19b and miR‐30c through SIRT3 activation, inhibits PAI‐1 levels. An association between PAI‐1, miR‐19b, and ‐30c was observed in plasma and microparticles isolated from patients suffering from acute coronary syndrome of various degrees.

  May 03, 2017   doi: 10.1002/jcp.25865   open full text
• Adenosine and adenosine receptors in the immunopathogenesis and treatment of cancer.
  Mohammad H. Kazemi, Sahar Raoofi Mohseni, Mohammad Hojjat‐Farsangi, Enayat Anvari, Ghasem Ghalamfarsa, Hamed Mohammadi, Farhad Jadidi‐Niaragh.
  Journal of Cellular Physiology. May 03, 2017
  Tumor cells overcome anti‐tumor responses in part through immunosuppressive mechanisms. There are several immune modulatory mechanisms. Among them, adenosine is an important factor which is generated by both cancer and immune cells in tumor microenvironment to suppress anti‐tumor responses. Two cell surface expressed molecules including CD73 and CD39 catalyze the generation of adenosine from adenosine triphosphate (ATP). The generation of adenosine can be enhanced under metabolic stress like tumor hypoxic conditions. Adenosine exerts its immune regulatory functions through four different adenosine receptors (ARs) including A1, A2A, A2B, and A3 which are expressed on various immune cells. Several studies have indicated the overexpression of adenosine generating enzymes and ARs in various cancers which was correlated with tumor progression. Since the signaling of ARs enhances tumor progression, their manipulation can be promising therapeutic approach in cancer therapy. Accordingly, several agonists and antagonists against ARs have been designed for cancer therapy. In this review, we will try to clarify the role of different ARs in the immunopathogenesis, as well as their role in the treatment of cancer. Adenosine is one of the important immunosuppressive factors in the tumor microenvironment. Therefore, modulation of adenosine receptors can be considered as the potent cancer therapeutic method.

  May 03, 2017   doi: 10.1002/jcp.25873   open full text
• MicroRNA106a regulates matrix metalloprotease 9 in a sirtuin‐1 dependent mechanism.
  Lincy Edatt, Ashutosh K. Maurya, Grace Raji, Haritha Kunhiraman, Sameer V. B. Kumar.
  Journal of Cellular Physiology. May 03, 2017
  Cellular migration is important during many physiological as well as pathological conditions and is regulated very tightly by an intricate network of signaling and effector molecules. One of the important players during cellular migration are matrix metalloproteases and their levels have been reported to be important in determining the cellular migratory properties during metastasis. MMPs and regulators of MMPs therefore, present themselves as potent candidates for manipulation, to control conditions where they get dysregulated. Micro RNAs are a group of micro regulators that can modulate expression of a gene through transcriptional and post transcriptional regulations. Owing to the fact that many microRNAs have already been reported to regulate MMPs and that miR106a, a member of oncomir17 family has been implicated in metastatic conditions, the present study intended to analyze if miR106a can regulate levels of MMP9, an important inducible matrix metalloproteinase. The results of the in vitro experiments demonstrated that under conditions of migration cells showed elevated levels of miR106a, which could regulate the expression of major MMP9 regulator, SIRT‐1. Decreased levels of SIRT1thus resulted in an increase in the expression and activity of MMP9. Over expression and mRNA stability studies carried out also suggested regulatory role of miR106a. The overall results thus suggested that the levels of miR106a gets modulated during cellular migration, causing a change in the levels of SIRT‐1 mRNA by affecting its stability and the levels of SIRT‐1 in turn can regulate the levels of MMP9. During cellular migration, the level of miRNA 106a is upregulated, which in turn regulates the levels of SIRT‐1 and further, SIRT‐1 modulates the levels of MMP‐9.

  May 03, 2017   doi: 10.1002/jcp.25870   open full text
• Soft tissue engineering with micronized‐gingival connective tissues.
  Sawako Noda, Yoshinori Sumita, Seigo Ohba, Hideyuki Yamamoto, Izumi Asahina.
  Journal of Cellular Physiology. May 03, 2017
  The free gingival graft (FGG) and connective tissue graft (CTG) are currently considered to be the gold standards for keratinized gingival tissue reconstruction and augmentation. However, these procedures have some disadvantages in harvesting large grafts, such as donor‐site morbidity as well as insufficient gingival width and thickness at the recipient site post‐treatment. To solve these problems, we focused on an alternative strategy using micronized tissue transplantation (micro‐graft). In this study, we first investigated whether transplantation of micronized gingival connective tissues (MGCTs) promotes skin wound healing. MGCTs (≤100 µm) were obtained by mincing a small piece (8 mm3) of porcine keratinized gingiva using the RIGENERA system. The MGCTs were then transplanted to a full skin defect (5 mm in diameter) on the dorsal surface of immunodeficient mice after seeding to an atelocollagen matrix. Transplantations of atelocollagen matrixes with and without micronized dermis were employed as experimental controls. The results indicated that MGCTs markedly promote the vascularization and epithelialization of the defect area 14 days after transplantation compared to the experimental controls. After 21 days, complete wound closure with low contraction was obtained only in the MGCT grafts. Tracking analysis of transplanted MGCTs revealed that some mesenchymal cells derived from MGCTs can survive during healing and may function to assist in wound healing. We propose here that micro‐grafting with MGCTs represents an alternative strategy for keratinized tissue reconstruction that is characterized by low morbidity and ready availability. Micronized‐gingival connective tissues promote the skin wound healing.

  May 03, 2017   doi: 10.1002/jcp.25871   open full text
• Profilin1 is expressed in osteocytes and regulates cell shape and migration.
  Wanting Lin, Yayoi Izu, Arayal Smriti, Makiri Kawasaki, Chantida Pawaputanon, Ralph T. Böttcher, Mercedes Costell, Keiji Moriyama, Masaki Noda, Yoichi Ezura.
  Journal of Cellular Physiology. May 03, 2017
  Osteocytes are the most abundant cells in bone and regulate bone metabolism in coordination with osteoblasts and osteoclasts. However, the molecules that control osteocytes are still incompletely understood. Profilin1 is an actin‐binding protein that is involved in actin polymerization. Osteocytes possess characteristic dendritic process formed based on actin cytoskeleton. Here, we examined the expression of profilin1 and its function in osteocytes. Profilin1 mRNA was expressed in osteocytic MLO‐Y4 cells and its levels were gradually increased along with the time in culture. With regard to functional aspect, knockdown of profilin1 by siRNA enhanced BMP‐induced increase in alkaline phosphatase expression levels in MLO‐Y4 cells. Profilin1 knockdown suppressed the levels of dendritic processes and migration of MLO‐Y4 cells. Since aging causes an increase in ROS in the body, we further examined the effects of hydrogen peroxide on the expression of profilin1. Hydrogen peroxide treatment increased the levels of profilin1 mRNA in MLO‐Y4 cells in contrast to the decline in alkaline phosphatase. Profilin1 was expressed not only in MLO‐Y4cells but also in the primary cultures of osteocytes. Importantly, profilin1 mRNA levels in primary cultures of osteocytes were higher than those in primary cultures of osteoblasts. To examine in vivo role of profilin1 in osteocytes, profilin1 was conditionally knocked out by using DMP1‐cre and profilin1 floxed mice. This conditional deletion of profilin1 specifically in osteocytes resulted in reduction in the levels of bone volume and bone mineral density. These data indicate that profilin1 is expressed in osteocytes and regulates cell shape, migration and bone mass. Profilin1 knockdown reduces dendritic process formation in osteocytic MLO‐Y4 cells.

  May 03, 2017   doi: 10.1002/jcp.25872   open full text
• Glucose adsorption to chitosan membranes increases proliferation of human chondrocyte via mammalian target of rapamycin complex 1 and sterol regulatory element‐binding protein‐1 signaling.
  Shun‐Fu Chang, Kuo‐Chin Huang, Chin‐Chang Cheng, Yu‐Ping Su, Ko‐Chao Lee, Cheng‐Nan Chen, Hsin‐I Chang.
  Journal of Cellular Physiology. May 03, 2017
  Osteoarthritis (OA) is currently still an irreversible degenerative disease of the articular cartilage. Recent, dextrose (d‐glucose) intraarticular injection prolotherapy for OA patients has been reported to benefit the chondrogenic stimulation of damaged cartilage. However, the detailed mechanism of glucose's effect on cartilage repair remains unclear. Chitosan, a naturally derived polysaccharide, has recently been investigated as a surgical or dental dressing to control breeding. Therefore, in this study, glucose was adsorbed to chitosan membranes (CTS‐Glc), and the study aimed to investigate whether CTS‐Glc complex membranes could regulate the proliferation of human OA chondrocytes and to explore the underlying mechanism. Human OA and SW1353 chondrocytes were used in this study. The experiments involving the transfection of cells used SW1353 chondrocytes. A specific inhibitor and siRNAs were used to investigate the mechanism underlying the CTS‐Glc‐regulated proliferation of human chondrocytes. We found that CTS‐Glc significantly increased the proliferation of both human OA and SW1353 chondrocytes comparable to glucose‐ or chitosan‐only stimulation. The role of mammalian target of rapamycin complex 1 (mTORC1) signaling, including mTOR, raptor, and S6k proteins, has been demonstrated in the regulation of CTS‐Glc‐increased human chondrocyte proliferation. mTORC1 signaling increased the expression levels of maturated SREBP‐1 and FASN and then induced the expressions of cell cycle regulators, that is, cyclin D, cyclin‐dependent kinase‐4 and ‐6 in human chondrocytes. This study elucidates the detailed mechanism behind the effect of CTS‐Glc complex membranes in promoting chondrocyte proliferation and proposes a possible clinical application of the CTS‐Glc complex in the dextrose intraarticular injection of OA prolotherapy in the future to attenuate the pain and discomfort of OA patients. The adsorption of glucose onto the chitosan membranes increased human chondrocyte proliferation by activating mTORC1 signaling and then induced the expression of mSREBP‐1/FASN/cell cycle regulators (cyclin D and CDK‐4/6).

  May 03, 2017   doi: 10.1002/jcp.25869   open full text
• Mitochondrial ROS‐induced ERK1/2 activation and HSF2‐mediated AT1R upregulation are required for doxorubicin‐induced cardiotoxicity.
  Chih‐Yang Huang, Jia‐Yi Chen, Chia‐Hua Kuo, Pei‐Ying Pai, Tsung‐Jung Ho, Tung‐Sheng Chen, Fu‐Jen Tsai, Vijaya V. Padma, Wei‐Wen Kuo, Chih‐Yang Huang.
  Journal of Cellular Physiology. May 03, 2017
  Doxorubicin (DOX), one useful chemotherapeutic agent, is limited in clinical use because of its serious cardiotoxicity. Growing evidence suggests that angiotensin receptor blockers (ARBs) have cardioprotective effects in DOX‐induced cardiomyopathy. However, the detailed mechanisms underlying the action of ARBs on the prevention of DOX‐induced cardiomyocyte cell death have yet to be investigated. Our results showed that angiotensin II receptor type I (AT1R) plays a critical role in DOX‐induced cardiomyocyte apoptosis. We found that MAPK signaling pathways, especially ERK1/2, participated in modulating AT1R gene expression through DOX‐induced mitochondrial ROS release. These results showed that several potential heat shock binding elements (HSE), which can be recognized by heat shock factors (HSFs), located at the AT1R promoter region. HSF2 markedly translocated from the cytoplasm to the nucleus when cardiomyocytes were damaged by DOX. Furthermore, the DNA binding activity of HSF2 was enhanced by DOX via deSUMOylation. Overexpression of HSF2 enhanced DOX‐induced cardiomyocyte cell death as well. Taken together, we found that DOX induced mitochondrial ROS release to activate ERK‐mediated HSF2 nuclear translocation and AT1R upregulation causing DOX‐damaged heart failure in vitro and in vivo. DOX induces mitochondrial ROS accumulation to activate ERK activation for HSF2‐mediated AT1R upregulation.

  May 03, 2017   doi: 10.1002/jcp.25905   open full text
• Environmental Temperature affect Physiology and Survival of Nanosecond Pulsed Electric Field Treated Cells.
  Shengyong Yin, Xudong Miao, Xueming Zhang, Xinhua Chen, Hao Wen.
  Journal of Cellular Physiology. May 03, 2017
  Nanosecond pulsed electric field (nsPEF) is a novel non‐thermal tumor ablation technique. However, how nsPEF affect cell physiology at different environmental temperature is still kept unknown. But this issue is of critical clinical practice relevance. This work aim to investigate how nsPEF treated cancer cells react to different environmental temperatures (0°C, 4°C, 25°C and 37°C). Their cell viability, apoptosis, mitochondrial membrane potential, and reactive oxygen species (ROS) were examined. Lower temperature resulted in higher apoptosis rate, decreased mitochondria membrane potential and increased ROS levels. Sucrose and N‐acetylcysteine (NAC) pre‐incubation inhibit ROS generation and increase cell survival, protecting nsPEF‐treated cells from low temperature‐caused cell death. This work provides an experimental basis for hypothermia and fluid transfusion during nsPEF ablation with anesthesia. This article is protected by copyright. All rights reserved

  May 03, 2017   doi: 10.1002/jcp.25984   open full text
• Pannexin hemichannels: A novel promising therapy target for oxidative stress related diseases.
  Jin Xu, Linxi Chen, Lanfang Li.
  Journal of Cellular Physiology. May 03, 2017
  Pannexins, which contain three subtypes: pannexin‐1, ‐2, and ‐3, are vertebrate glycoproteins that form non‐junctional plasma membrane intracellular hemichannels via oligomerization. Oxidative stress refers to an imbalance of the generation and elimination of reactive oxygen species (ROS). Studies have shown that elevated ROS levels are pivotal in the development of a variety of diseases. Recent studies indicate that the occurrence of these oxidative stress related diseases is associated with pannexin hemichannels. It is also reported that pannexins regulate the production of ROS which in turn may increase the opening of pannexin hemichannels. In this paper, we review recent researches about the important role of pannexin hemichannels in oxidative stress related diseases. Thus, pannexin hemichannels, novel therapeutic targets, hold promise in managing oxidative stress related diseases such as the tumor, inflammatory bowel diseases (IBD), pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), cardiovascular disease, insulin resistance (IR), and neural degeneration diseases. In this review, we find that there is intimate relationship between pannexin hemichannels and oxidative stress, thus we discuss possible mechanism of pannexin hemichannels on oxidative stress related diseases.

  May 03, 2017   doi: 10.1002/jcp.25906   open full text
• Roles of platelet‐derived growth factor in vascular calcification.
  Liu Ouyang, Kun Zhang, Jie Chen, Jingfeng Wang, Hui Huang.
  Journal of Cellular Physiology. May 03, 2017
  Vascular calcification (VC) is prevalent in aging, and patients with hypertension, chronic kidney disease (CKD) or diabetes. VC is regarded as an active and complex process that involves multiple mechanisms responsible for calcium deposition in vessel wall. In light of the complicated pathogenesis of VC, effective therapy for ameliorating VC is limited. Thus, it is urgent to explore the potential mechanisms and find new targets for the therapy of VC. Platelet‐derived growth factor (PDGF), a potent mitogen and chemoattractant, has been found to disturb the vascular homeostasis by inducing inflammation, oxidative stress and phenotype transition, all of which accelerate the process of VC. The aim of current review is to present a review about the roles of PDGF in affecting VC and to establish a potential target for treating VC. This article is protected by copyright. All rights reserved

  May 03, 2017   doi: 10.1002/jcp.25985   open full text
• Reduced glucose‐induced insulin secretion in low‐protein‐fed rats is associated with altered pancreatic islets redox status.
  Ana Paula G. Cappelli, Claudio C. Zoppi, Leonardo R. Silveira, Thiago M. Batista, Flávia M. Paula, Priscilla M. R. da Silva, Alex Rafacho, Helena C. Barbosa‐Sampaio, Antonio C. Boschero, Everardo M. Carneiro.
  Journal of Cellular Physiology. May 03, 2017
  In the present study, we investigated the relationship between early life protein malnutrition‐induced redox imbalance, and reduced glucose‐stimulated insulin secretion. After weaning, male Wistar rats were submitted to a normal‐protein‐diet (17%‐protein, NP) or to a low‐protein‐diet (6%‐protein, LP) for 60 days. Pancreatic islets were isolated and hydrogen peroxide (H2O2), oxidized (GSSG) and reduced (GSH) glutathione content, CuZn‐superoxide dismutase (SOD1), glutathione peroxidase (GPx1) and catalase (CAT) gene expression, as well as enzymatic antioxidant activities were quantified. Islets that were pre‐incubated with H2O2 and/or N‐acetylcysteine, were subsequently incubated with glucose for insulin secretion measurement. Protein malnutrition increased CAT mRNA content by 100%. LP group SOD1 and CAT activities were 50% increased and reduced, respectively. H2O2 production was more than 50% increased whereas GSH/GSSG ratio was near 60% lower in LP group. Insulin secretion was, in most conditions, approximately 50% lower in LP rat islets. When islets were pre‐incubated with H2O2 (100 μM), and incubated with glucose (33 mM), LP rats showed significant decrease of insulin secretion. This effect was attenuated when LP islets were exposed to N‐acetylcysteine. Our data suggest that short‐term early life protein malnourishment induces reduction in redox GIIS signaling only at extreme oxidant challenges, elicited here by high H2O2 pre‐incubation followed by elevated glucose concentration incubation, probably due to lower antioxidant scavenging capacity.

  May 03, 2017   doi: 10.1002/jcp.25908   open full text
• Probiotics are a good choice in remission of inflammatory bowel diseases: A meta analysis and systematic review.
  Mahboube Ganji‐Arjenaki, Mahmoud Rafieian‐Kopaei.
  Journal of Cellular Physiology. May 03, 2017
  Altered gut bacteria and bacterial metabolic pathways are two important factors in initiation and progression of inflammatory bowel disease (IBD). However, efficacy of probiotics in remission of patients with IBD has not been characterized. This study was performed on the studies that specifically assessed the efficacy of probiotics in attaining clinical response on patients with various types of IBD. The efficacy of variant species of probiotics in different conditions and the influence of study quality in outcomes of randomized controlled trials (RCTs) were also assessed. The RCTs were collected by searching in MEDLINE Web of Science and Google scholar. Then all studies were abstracted in abstraction form and the outcomes were analyzed with fixed‐effect and mixed‐effect models for assessment of efficacy of variant species of probiotics in subgroups of IBDs. Analysis of 9 trials showed that probiotics had not significant effect on Crohn's disease (CD) (p = 0.07) but analysis of 3 trials in children with IBD revealed a significant advantage (p < 0.01). Analysis of 18 trials revealed that probiotics in patients with Ulcerative colitis (UC) in different conditions have significant effects (p = 0.007). VSL#3 probiotics in patients with UC had significant effect (p < 0.01). Combination of Lactobacillus probiotic, prebiotics had significant effect (p = 0.03) only in patients with UC. Combination of Saccharomyces boulardii, Lactobacillus, and VSL#3 probiotics in CD had also a trend for efficiency (p = 0.057). In children with IBD, the combination of Lactobacillus with VSL#3 probiotics had significant effect (p < 0.01). Probiotics are beneficial in IBD, especially the combination ones in UC.

  May 03, 2017   doi: 10.1002/jcp.25911   open full text
• Autophagy mediates cytotoxicity of human colorectal cancer cells treated with garcinielliptone FC.
  Shen‐Jeu Won, Cheng‐Hsin Yen, Ting‐Yu Lin, Ya‐Fen Jiang‐Shieh, Chun‐Nan Lin, Jyun‐Ti Chen, Chun‐Li Su.
  Journal of Cellular Physiology. May 03, 2017
  The tautomeric pair of garcinielliptone FC (GFC) is a novel tautomeric pair of polyprenyl benzophenonoid isolated from the pericarps of Garcinia subelliptica Merr. (G. subelliptica, Clusiaceae), a tree with abundant sources of polyphenols. Our previous report demonstrated that GFC induced apoptosis on various types of human cancer cell lines including chemoresistant human colorectal cancer HT‐29 cells. In the present study, we observed that many autophagy‐related genes in GFC‐treated HT‐29 cells were up‐ and down‐regulated using a cDNA microarray containing oncogenes and kinase genes. GFC‐induced autophagy of HT‐29 cells was confirmed by observing the formation of acidic vesicular organelles, LC3 puncta, and double‐membrane autophagic vesicles using flow cytometry, confocal microscopy, and transmission electron microscopy, respectively. Inhibition of AKT/mTOR/P70S6K signaling as well as formation of Atg5‐Atg12 and PI3K/Beclin‐1 complexes were observed using Western blot. Administration of autophagy inhibitor (3‐methyladenine and shRNA Atg5) and apoptosis inhibitor Z‐VAD showed that the GFC‐induced autophagy was cytotoxic form and GFC‐induced apoptosis enhanced GFC‐induced autophagy. Our data suggest the involvement of autophagy and apoptosis in GFC‐induced anticancer mechanisms of human colorectal cancer. The tautomeric pair of garcinielliptone FC (GFC) is a novel tautomeric pair of polyprenyl benzophenonoid isolated from the pericarps of Garcinia subelliptica Merr. (G. subelliptica, Clusiaceae), a tree with abundant sources of polyphenols. GFC‐induced autophagy was cytotoxic form and GFC‐induced apoptosis enhanced GFC‐induced autophagy.

  May 03, 2017   doi: 10.1002/jcp.25910   open full text
• Knockout of ATG5 leads to malignant cell transformation and resistance to Src family kinase inhibitor PP2.
  Sung‐Hee Hwang, Byeal‐I Han, Michael Lee.
  Journal of Cellular Physiology. May 03, 2017
  Autophagy can either promote or inhibit cell death in different cellular contexts. In this study, we investigated the role of autophagy in ATG5 knockout (KO) cell line established using CRISPR/Cas9 system. In ATG5 KO cells, RT‐PCR and immunoblot of LC3 confirmed the functional gene knockout. We found that knockout of ATG5 significantly increased proliferation of NIH 3T3 cells. In particular, autophagy deficiency enhanced susceptibility to cellular transformation as determined by an in vitro clonogenic survival assay and a soft agar colony formation assay. We also found that ATG5 KO cells had a greater migration ability as compared to wild‐type (WT) cells. Moreover, ATG5 KO cells were more resistant to treatment with a Src family tyrosine kinase inhibitor (PP2) than WT cells were. Cyto‐ID Green autophagy assay revealed that PP2 failed to induce autophagy in ATG5 KO cells. PP2 treatment decreased the percentage of cells in the S and G2/M phases among WT cells but had no effect on cell cycle distribution of ATG5 KO cells, which showed a high percentage of cells in the S and G2/M phases. Additionally, the proportion of apoptotic cells significantly decreased after treatment of ATG5 KO cells with PP2 in comparison with WT cells. We found that expression levels of p53 were much higher in ATG5 KO cells. The ATG5 KO seems to lead to compensatory upregulation of the p53 protein because of a decreased apoptosis rate. Taken together, our results suggest that autophagy deficiency can lead to malignant cell transformation and resistance to PP2. ATG5 knockout cells show morphological transformation and anchorage‐independent growth‐ ATG5 KO cells have a lower proportion of the apoptotic cell population as a way to escape cell death in response to PP2.

  May 03, 2017   doi: 10.1002/jcp.25912   open full text
• Modulating tumor hypoxia by nanomedicine for effective cancer therapy.
  Rana Jahanban‐Esfahlan, Miguel de la Guardia, Delshad Ahmadi, Bahman Yousefi.
  Journal of Cellular Physiology. May 03, 2017
  Hypoxia, a characteristic feature of tumors, is indispensable to tumor angiogenesis, metastasis, and multi drug resistance. Hypoxic avascular regions, deeply embedded inside the tumors significantly hinder delivery of therapeutic agents. The low oxygen tension results in resistance to the current applied anti‐cancer therapeutics including radiotherapy, chemotherapy, and photodynamic therapy, the efficacy of which is firmly tied to the level of tumor oxygen supply. However, emerging data indicate that nanocarriers/nanodrugs can offer substantial benefits to improve the efficacy of current therapeutics, through modulation of tumor hypoxia. This review aims to introduce the most recent advances made in nanocarrier mediated targeting of tumor hypoxia. The first part is dedicated to the approaches by which nanocarriers could be designed to target/leverage hypoxia. These approaches include i) inhibiting Hypoxia Inducer Factor (HIF‐1α); ii) hypoxia activated prodrugs/linkers; and iii) obligate anaerobe mediated targeting of tumor hypoxia. The second part, details novel nanosystems proposed to modulate tumor hypoxia through tumor oxygenation. These methods seek to lessen tumor hypoxia through vascular normalization, or reoxygenation therapy. The reoxygenation of tumor could be accomplished by: i) generation of oxygen filled nanocarriers; ii) natural/artificial oxygen nanocarriers; and iii) oxygen generators. The efficacy of each approach and their potential in cancer therapy is further discussed. Hypoxic avascular regions, deeply embedded inside the tumors significantly hinder delivery of therapeutic agents. Hypoxia could be accomplished by developing HIF‐1a and VEGF inhibitors that are now successfully integrated into the oncology practice. Incorporation of nanomedicine in targeting tumor hypoxia could dramatically enhance potential of anti‐cancer therapeutics.

  May 03, 2017   doi: 10.1002/jcp.25859   open full text
• Surface Functionalized Magnetic Nanoparticles Shift Cell Behavior with On/Off Magnetic Fields.
  Seongbeom Jeon, Ramesh Subbiah, Taufik Bonaedy, Seyoung Van, Kwideok Park, Kyusik Yun.
  Journal of Cellular Physiology. May 02, 2017
  Magnetic nanoparticles (MNPs) are used as contrast agents and targeted drug delivery systems (TDDS) due to their favorable size, surface charge, and magnetic properties. Unfortunately, the toxicity associated with MNPs limits their biological applications. Surface functionalization of MNPs with selective polymers alters the surface chemistry to impart better biocompatibility. We report the preparation of surface functionalized MNPs using iron oxide NPs (MNPs), poly (lactic‐co‐glycolic acid) (PLGA) and sodium alginate via co‐precipitation, emulsification, and electro‐spraying, respectively. The NPs are in the nanosize range and negatively charged. Morphological and structural analyses affirm the surface functionalized nanostructure of the NPs. The surface functionalized MNPs are biocompatible, and demonstrate enhanced intracellular delivery under an applied magnetic field (H), which evinces the targeting ability of MNPs. After NP treatment, the physico‐mechanical properties of fibroblasts are decided by the selective MNP uptake under “on” or “off” magnetic field conditions. We envision potential use of biocompatible surface functionalized MNP for intracellular‐, targeted‐DDS, imaging, and for investigating cellular mechanics. This article is protected by copyright. All rights reserved

  May 02, 2017   doi: 10.1002/jcp.25980   open full text
• The Skeletal Cell‐Derived Molecule Sclerostin Drives Bone Marrow Adipogenesis.
  Heather Fairfield, Carolyne Falank, Elizabeth Harris, Victoria Demambro, Michelle McDonald, Jessica A Pettit, Sindhu T Mohanty, Peter Croucher, Ina Kramer, Michaela Kneissel, Clifford J. Rosen, Michaela R. Reagan.
  Journal of Cellular Physiology. April 29, 2017
  The bone marrow niche is a dynamic and complex microenvironment that can both regulate, and be regulated by the bone matrix. Within the bone marrow (BM), mesenchymal stromal cell (MSC) precursors reside in a multi‐potent state and retain the capacity to differentiate down osteoblastic, adipogenic, or chondrogenic lineages in response to numerous biochemical cues. These signals can be altered in various pathological states including, but not limited to, osteoporotic‐induced fracture, systemic adiposity, and the presence of bone‐homing cancers. Herein we provide evidence that signals from the bone matrix (osteocytes) determine marrow adiposity by regulating adipogenesis in the bone marrow. Specifically, we found that physiologically relevant levels of Sclerostin (SOST), which is a Wnt‐inhibitory molecule secreted from bone matrix‐embedded osteocytes, can induce adipogenesis in 3T3‐L1 cells, mouse ear‐ and BM‐derived MSCs, and human BM‐derived MSCs. We demonstrate that the mechanism of SOST induction of adipogenesis is through inhibition of Wnt signaling in pre‐adipocytes. We also demonstrate that a decrease of sclerostin in vivo, via both genetic and pharmaceutical methods, significantly decreases bone marrow adipose tissue (BMAT) formation. Overall, this work demonstrates a direct role for SOST in regulating fate determination of BM‐adipocyte progenitors. This provides a novel mechanism for which BMAT is governed by the local bone microenvironment, which may prove relevant in the pathogenesis of certain diseases involving marrow adipose. Importantly, with anti‐sclerostin therapy at the forefront of osteoporosis treatment and a greater recognition of the role of BMAT in disease, these data are likely to have important clinical implications.This article is protected by copyright. All rights reserved

  April 29, 2017   doi: 10.1002/jcp.25976   open full text
• Interplay between miRNAs and human diseases.
  Prosenjit Paul, Anindya Chakraborty, Debasree Sarkar, Malobika Langthasa, Musfhia Rahman, Minakshi Bari, RK. Sanamacha Singha, Arup Kumar Malakar, Supriyo Chakraborty.
  Journal of Cellular Physiology. April 27, 2017
  MicroRNAs (miRNAs) are endogenous, non‐coding RNAs, which have evoked a great deal of interest due to their importance in many aspects of homeostasis and diseases. MicroRNAs are stable and are essential components of gene regulatory networks. They play a crucial role in healthy individuals and their dysregulations have also been implicated in a wide range of diseases, including diabetes, cardiovascular disease, kidney disease, and cancer. This review summarized the current understanding of interactions between miRNAs and different diseases and their role in disease diagnosis and therapy. This review summarized the current understanding of interactions between miRNAs and different diseases and their role in disease diagnosis and therapy.

  April 27, 2017   doi: 10.1002/jcp.25854   open full text
• Effect of stress on human biology: Epigenetics, adaptation, inheritance, and social significance.
  David T. Denhardt.
  Journal of Cellular Physiology. April 27, 2017
  I present a brief introduction to epigenetics, focused primarily on methylation of the genome and various regulatory RNAs, modifications of associated histones, and their importance in enabling us to adapt to real and changing environmental, developmental, and social circumstances. Following this is a more extensive overview of how it impacts our inheritance, our entire life (which changes as we age), and how we interact with others. Throughout, I emphasize the critical influence that stress, of many varieties exerts, via epigenetic means, on much of how we live and survive, mostly in the brain. I end with a short section on multigenerational transmission, drugs, and the importance of both social life and early life experiences in the development of adult diseases. There will be nothing about cancer. Although epigenetics is critical in that field, it is a whole different cobweb of complications (some involving stress). This review focuses on the impact of stress of many kinds on human physiology. It discusses DNA and histone modifications and regulation by various forms of RNA. It ends with a critical analysis of the impact of drugs and social interactions on the individual and transgenerational inheritance.

  April 27, 2017   doi: 10.1002/jcp.25837   open full text
• IL‐10 expression in macrophages from neonates born from obese mothers is suppressed by IL‐4 and LPS/INFγ.
  Francisca Cifuentes‐Zúñiga, Viviana Arroyo‐Jousse, Gustavo Soto‐Carrasco, Paola Casanello, Ricardo Uauy, Bernardo J. Krause, José A. Castro‐Rodríguez.
  Journal of Cellular Physiology. April 27, 2017
  Obese women offspring have a higher risk of developing chronic diseases associated with an altered immune function. We aim to determine, in neonatal monocyte‐derived macrophages, whether maternal obesity is associated with an altered expression and DNA methylation of pro‐ and anti‐inflammatory genes, along with a higher pro‐inflammatory response. Cord blood from newborns of obese (Ob) and lean (control) women were obtained at delivery. Monocytes were isolated and differentiated into macrophages, in which M1 (LPS/IFNγ) and M2 (IL‐4) polarization were assayed. The mRNA levels for TNFα, IL‐1β, IL‐12A, IL‐12B, IL‐10, and IL‐4R were quantified by qPCR and the DNA methylation of candidate genes determined by pyrosequencing. Results: Ob‐monocytes had decreased levels of mRNA for pro‐inflammatory cytokines IL‐1β, IL‐10, and IL‐12B compared with controls. Conversely, Ob‐macrophages showed increased levels of mRNA for TNFα, IL‐4R, and IL‐10 compared with controls. M1 response was comparable between both groups, characterized by an important induction of TNFα and IL‐1β. In response to an M2 stimulus, control macrophages showed a decreased expression of inflammatory mediators while Ob‐macrophages had an additional suppression of the anti‐inflammatory mediator IL‐10. Changes in IL‐1β (monocytes) and IL‐10 (macrophages) in Ob‐monocytes were paralleled by changes in their promoter DNA methylation in fetal monocytes. These results suggest that monocyte‐derived macrophages from obese newborns show a basal anti‐inflammatory phenotype with an unbalanced response to M1 and M2 polarization stimuli. The presence of changes in DNA methylation of key inflammatory genes in neonatal monocytes suggests an intrauterine programing of immune function by maternal obesity.

  April 27, 2017   doi: 10.1002/jcp.25845   open full text
• A prognostic 4‐gene expression signature for squamous cell lung carcinoma.
  Jun Li, Jing Wang, Yanbin Chen, Lijie Yang, Sheng Chen.
  Journal of Cellular Physiology. April 27, 2017
  Squamous cell lung carcinoma (SQCLC), a common and fatal subtype of lung cancer, caused lots of mortalities and showed different outcomes in prognosis. This study was to screen key genes and to figure a prognostic signature to cluster the patients with SQCLC. RNA‐Seq data from 550 patients with SQCLC were downloaded from The Cancer Genome Atlas (TCGA). Genetically changed genes were identified and analyzed in univariate survival analysis. Genes significantly influencing prognosis were selected with frequency higher than 100 in lasso regression. Meanwhile, area under the curve (AUC) values and hazard ratios (HR) for seed genes were obtained with R Language. Functional enrichment analysis was performed and clustering effectiveness of the selected common gene set was analyzed with Kaplan–Meier. Finally, the stability and validity of the optimal clustering model were verified. A total of 7,222 genetically changed genes were screened, including 1,045 ones with p < 0.05, 1,746, p < 0.1, and 2,758, p < 0.2. The common gene sets with more than 100 frequencies were 14‐Genes, 10‐Genes and 6‐Genes. Genes with p < 0.05 participated in positive regulation of ERK1 and ERK2 cascade, angiogenesis, platelet degranulation, cell–matrix adhesion, extracellular matrix organization, macrophage activation, and so on. A four‐gene clustering model in 14‐Genes (DPPA, TTTY16, TRIM58, HKDC1, ZNF589, ALDH7A1, LINC01426, IL19, LOC101928358, TMEM92, HRASLS, JPH1, LOC100288778, GCGR) was verified as the optimal. The discovery of four‐gene clustering model in 14‐Genes can cluster the patient samples effectively. This model would help predict the outcomes of patients with SQCLC then improve the treatment strategies. A four‐gene clustering model in 14‐Genes (DPPA, TTTY16, TRIM58, HKDC1, ZNF589, ALDH7A1, LINC01426, IL19, LOC101928358, TMEM92, HRASLS, JPH1, LOC100288778, GCGR) was verified as the optimal. The discovery of four‐gene clustering model in 14‐Genes can cluster the patient samples effectively. This model would help predict the outcomes of patients with SQCLC then improve the treatment strategies.

  April 27, 2017   doi: 10.1002/jcp.25846   open full text
• Molecular pathogenesis in chronic obstructive pulmonary disease and therapeutic potential by targeting AMP‐activated protein kinase.
  Zhihui Zhang, Xiaoyu Cheng, Li Yue, Wenhui Cui, Wencheng Zhou, Jian Gao, Hongwei Yao.
  Journal of Cellular Physiology. April 27, 2017
  Chronic obstructive pulmonary disease (COPD) is the third leading cause of death worldwide, which is characterized by a persistent airflow limitation caused by chronic inflammatory responses to noxious particles or gases. Cigarette smoke and environmental pollutions are major etiological factors for causing COPD. It has been shown that cigarette smoking causes abnormal inflammatory responses, cellular senescence, mitochondrial dysfunction and metabolic dysregulation, suggesting their involvement in the development of COPD. Although the medical care and treatment have advanced, there are no effective therapies to stop or reverse lung destruction in COPD/emphysema. AMP‐activated protein kinase (AMPK) is a serine threonine kinase with α, β, and γ subunits that are highly conserved through evolution. AMPK has been shown to regulate bioenergetics, inflammatory responses, senescence, and metabolism. This review focused on the updated understanding of molecular pathogenesis of COPD, and highlighted the crucial roles of AMPK in lung abnormalities as well as discussed the potential therapeutics of AMPK activators in preventing and halting the progression of COPD. Cigarette smoke as well as other gases and particles cause inflammatory responses, senescence, mitochondrial dysfunction, and metabolic dysregulation, leading to lung destruction and subsequently COPD/emphysema. AMPK is reduced by cigarette smoke and in lungs of COPD patients, and AMPK activators protect against aforementioned cellular processes and pulmonary emphysema.

  April 27, 2017   doi: 10.1002/jcp.25844   open full text
• Downregulation of methyltransferase Dnmt2 results in condition‐dependent telomere shortening and senescence or apoptosis in mouse fibroblasts.
  Anna Lewinska, Jagoda Adamczyk‐Grochala, Ewa Kwasniewicz, Maciej Wnuk.
  Journal of Cellular Physiology. April 27, 2017
  Dnmt2 is a highly conserved methyltransferase of uncertain biological function(s). As Dnmt2 was considered as a driver of fruit fly longevity and a modulator of stress response, we decided to evaluate the role of Dnmt2 during stress‐induced premature senescence in NIH3T3 mouse fibroblasts. Stable knockdown of Dnmt2 resulted in hydrogen peroxide‐mediated sensitivity and apoptosis, whereas in the control conditions, senescence was induced. Cellular senescence was accompanied by elevated levels of p53 and p21, decreased telomere length and telomerase activity, increased production of reactive oxygen species and protein carbonylation, and DNA damage. Dnmt2 silencing also promoted global DNA and RNA hypermethylation, and upregulation of methyltransferases, namely Dnmt1, Dnmt3a, and Dnmt3b. Taken together, we show for the first time that Dnmt2 may promote lifespan in the control conditions and survival during stress conditions in mouse fibroblasts. Dnmt2 silencing results in cellular senescence and apoptosis in the control conditions and after HP treatment, respectively. Senescence was accompanied by mild oxidative stress, DNA damage and telomere shortening, whereas Dnmt2‐silenced fibroblasts were more prone to ROS‐induced apoptotic cell death after HP stimulation.

  April 27, 2017   doi: 10.1002/jcp.25848   open full text
• Sphingolipids metabolism in the salivary glands of rats with obesity and streptozotocin induced diabetes.
  Marta Garbowska, Bartłomiej Łukaszuk, Agnieszka Mikłosz, Igor Wróblewski, Krzysztof Kurek, Lucyna Ostrowska, Adrian Chabowski, Małgorzata Żendzian‐Piotrowska, Anna Zalewska.
  Journal of Cellular Physiology. April 27, 2017
  Diabetes is considered a major public health problem affecting millions of individuals worldwide. Remarkably, scientific reports regarding salivary glands sphingolipid metabolism in diabetes are virtually non‐existent. This is odd given the well‐established link between the both in other tissues (e.g., skeletal muscles, liver) and the key role of these glands in oral health preservation. The aim of this paper is to examine sphingolipids metabolism in the salivary glands in (pre)diabetes (evoked by high fat diet feeding or streptozotocin). Wistar rats were allocated into three groups: control, HFD‐, or STZ‐diabetes. The content of major sphingolipid classes in the parotid (PSG) and submandibular (SMSG) glands was assessed via chromatography. Additionally, Western blot analyses were employed for the evaluation of key sphingolipid signaling pathway enzyme levels. No changes in ceramide content in the PSG were found, whereas an increase in ceramide concentration for SMSG of the STZ group was observed. This was accompanied by an elevation in SPT1 level. Probably also sphingomyelin hydrolysis was increased in the SMSG of the STZ‐diabetic rats, since we observed a significant drop in the amount of SM. PSG and SMSG respond differently to (pre)diabetes, with clearer pattern presented by the later gland. An activation of sphingomyelin signaling pathway was observed in the course of STZ‐diabetes, that is, metabolic condition with rapid onset/progression. Whereas, chronic HFD lead to an inhibition of sphingomyelin signaling pathway in the salivary glands (manifested in an inhibition of ceramide de novo synthesis and accumulation of S1P). In this study we explored sphingolipid metabolism in the rats salivary glands in both diet induced insulin resistance and type 1 diabetic condition. Based on our results we conclude that the SMSG are characterized by somewhat greater metabolic activity in comparison to PSG. Another interesting and novel finding of our study is an activation of sphingomyelin signaling pathway observed only in the course of type 1 diabetes, which is a metabolic condition with rapid onset and progression.

  April 27, 2017   doi: 10.1002/jcp.25939   open full text
• The SGK1 inhibitor SI113 induces autophagy, apoptosis, and endoplasmic reticulum stress in endometrial cancer cells.
  Domenico Conza, Paola Mirra, Gaetano Calì, Teresa Tortora, Luigi Insabato, Francesca Fiory, Silvia Schenone, Rosario Amato, Francesco Beguinot, Nicola Perrotti, Luca Ulianich.
  Journal of Cellular Physiology. April 27, 2017
  Endometrial cancer is often characterized by PI3K/AKT pathway deregulation. Recently it has been suggested that SGK1, a serine/threonine protein kinase that shares structural and functional similarities with the AKT family, might play a role in cancer, since its expression and/or activity has been found to be deregulated in different human tumors. However, the role of SGK1 in endometrial cancer has been poorly investigated. Here, we show that SGK1 expression is increased in tissue specimens from neoplastic endometrium. The SGK1 inhibitor SI113 induced a significant reduction of endometrial cancer cells viability, measured by the (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide assay. This effect was associated to the increase of autophagy, as revealed by the increase of the markers LC3B‐II and beclin I, detected by both immunofluorescence and western blot analysis. SI113 treatment caused also apoptosis of endometrial cancer cells, evidenced by the cleavage of the apoptotic markers PARP and Caspase‐9. Intriguingly, these effects were associated to the induction of endoplasmic reticulum stress markers GRP78 and CHOP evaluated by both Real‐Time RT‐PCR and Western Blot analysis. Increased expression of SGK1 in endometrial cancer tissues suggest a role for SGK1 in this type of cancer, as reported for other malignancies. Moreover, the efficacy of SI113 in affecting endometrial cancer cells viability, possibly via endoplasmic reticulum stress activation, identifies SGK1 as an attractive molecular target for new tailored therapeutic intervention for the treatment of endometrial cancer. SGK1 expression is increased in endometrial cancer SI113, an inhibitor of SGK1, induces endoplasmic reticulum stress, autophagy, and apoptosis in endometrial cancer cells.

  April 27, 2017   doi: 10.1002/jcp.25850   open full text
• Role of PARP activity in lung cancer‐induced cachexia: Effects on muscle oxidative stress, proteolysis, anabolic markers, and phenotype.
  Alba Chacon‐Cabrera, Mercè Mateu‐Jimenez, Klaus Langohr, Clara Fermoselle, Elena García‐Arumí, Antoni L. Andreu, Jose Yelamos, Esther Barreiro.
  Journal of Cellular Physiology. April 27, 2017
  Strategies to treat cachexia are still at its infancy. Enhanced muscle protein breakdown and ubiquitin‐proteasome system are common features of cachexia associated with chronic conditions including lung cancer (LC). Poly(ADP‐ribose) polymerases (PARP), which play a major role in chromatin structure regulation, also underlie maintenance of muscle metabolism and body composition. We hypothesized that protein catabolism, proteolytic markers, muscle fiber phenotype, and muscle anabolism may improve in respiratory and limb muscles of LC‐cachectic Parp‐1‐deficient (Parp‐1−/−) and Parp‐2−/− mice. In diaphragm and gastrocnemius of LC (LP07 adenocarcinoma) bearing mice (wild type, Parp‐1−/−, and Parp‐2−/−), PARP activity (ADP‐ribose polymers, pADPr), redox balance, muscle fiber phenotype, apoptotic nuclei, tyrosine release, protein ubiquitination, muscle‐specific E3 ligases, NF‐κB signaling pathway, markers of muscle anabolism (Akt, mTOR, p70S6K, and mitochondrial DNA) were evaluated along with body and muscle weights, and limb muscle force. Compared to wild type cachectic animals, in both respiratory and limb muscles of Parp‐1−/− and Parp‐2−/− cachectic mice: cancer induced‐muscle wasting characterized by increased PARP activity, protein oxidation, tyrosine release, and ubiquitin‐proteasome system (total protein ubiquitination, atrogin‐1, and 20S proteasome C8 subunit) were blunted, the reduction in contractile myosin and atrophy of the fibers was attenuated, while no effects were seen in other structural features (inflammatory cells, internal or apoptotic nuclei), and markers of muscle anabolism partly improved. Activation of either PARP‐1 or ‐2 is likely to play a role in muscle protein catabolism via oxidative stress, NF‐κB signaling, and enhanced proteasomal degradation in cancer‐induced cachexia. Therapeutic potential of PARP activity inhibition deserves attention. The study findings imply that activation of either PARP‐1 or ‐2 is likely to play a relevant role in muscle protein catabolism via oxidative stress, NF‐κB signaling, and enhanced proteasomal degradation in cancer‐induced cachexia. Despite the potential limitations of PARP activity inhibition, since it plays a key role in DNA maintenance, exploration of its reliability, and likely applicability as a possible target for future therapeutic interventions deserves attention in models of cancer‐induced cachexia.

  April 27, 2017   doi: 10.1002/jcp.25851   open full text
• Polydatin impairs mitochondria fitness and ameliorates podocyte injury by suppressing Drp1 expression.
  Zheng Ni, Liang Tao, Xu Xiaohui, Zhao Zelin, Liu Jiangang, Song Zhao, Huo Weikang, Xu Hongchao, Wang Qiujing, Li Xin.
  Journal of Cellular Physiology. April 27, 2017
  Polydatin (PD), a resveratrol glycoside, has been shown to protect renal function in diabetic nephropathy (DN), but the underlying molecular mechanism remains unclear. This study demonstrates that PD stabilize the mitochondrial morphology and attenuate mitochondrial malfunction in both KKAy mice and in hyperglycemia (HG)‐induced MPC5 cells. We use Western blot analysis to demonstrate that PD reversed podocyte apoptosis induced by HG via suppressing dynamin‐related protein 1 (Drp1). This effect may depend on the ability of PD to inhibit the generation of cellular reactive oxygen species (ROS). In conclusion, we demonstrate that PD may be therapeutically useful in DN, and that, podocyte apoptosis induced by HG can be reversed by PD through suppressing Drp1 expression. Our findings provide new insights into the pathogenic process of HG‐induced podocyte injury and also identify a new therapeutic target of ROS/Drp1/mitochondrial fission/apoptosis pathway for diabetic nephropathy.

  April 27, 2017   doi: 10.1002/jcp.25943   open full text
• VSL#3 probiotic differently influences IEC‐6 intestinal epithelial cell status and function.
  Benedetta Cinque, Cristina La Torre, Francesca Lombardi, Paola Palumbo, Zoran Evtoski, Silvano Jr Santini, Stefano Falone, Annamaria Cimini, Fernanda Amicarelli, Maria Grazia Cifone.
  Journal of Cellular Physiology. April 25, 2017
  The data here reported introduce the wound‐healing assay as a tool for testing probiotics aimed at protecting gastrointestinal mucosal surfaces and to verify the consistency of their manufacturing. At the scope, we compared the in vitro effects of two multi‐strain high concentration formulations both commercialized under the same brand VSL#3 but sourced from different production sites (USA and Italy) on a non‐transformed small‐intestinal epithelial cell line, IEC‐6. The effects on cellular morphology, viability, migration, and H2O2‐induced damage, were assessed before and after the treatment with both VSL#3 formulations. While the USA‐sourced product (“USA‐made”) VSL#3 did not affect monolayer morphology and cellular density, the addition of bacteria from the Italy‐derived product (“Italy‐made”) VSL#3 caused clear morphological cell damage and strongly reduced cellularity. The treatment with “USA‐made” lysate led to a higher rate of wounded monolayer healing, while the addition of “Italy‐made” bacterial lysate did not influence the closure rate as compared to untreated cells. While lysates from “USA‐made” VSL#3 clearly enhanced the formation of elongated and aligned stress fibers, “Italy‐made” lysates had not similar effect. “USA‐made” lysate was able to cause a total inhibition of H2O2‐induced cytotoxic effect whereas “Italy‐made” VSL#3 lysate was unable to protect IEC‐6 cells from H2O2‐induced damage. ROS generation was also differently influenced, thus supporting the hypotesis of a protective action of “USA‐made” VSL#3 lysates, as well as the idea that “Italy‐made” formulation was unable to prevent significantly the H2O2‐induced oxidative stress. The data here reported introduce the wound‐healing assay as a tool for testing probiotics aimed at protecting gastrointestinal mucosal surfaces and to verify the consistency of their manufacturing. At the scope, we compared the in vitro effects of two multi‐strain high concentration formulations both commercialized under the same brand VSL#3 but sourced from different production sites (USA and Italy) on a non‐transformed small‐intestinal epithelial cell line, IEC‐6. Our results suggest that all the cellular parameters analyzed on a rat intestinal epithelial cell line (IEC‐6)—cell viability and death, wounded monolayer healing ability and protection against H2O2‐induced cellular injury—appear to be differently influenced by the “Italy‐made” VSL#3 when compared to the “USA‐made” VSL#3, thus reconfirming previous reports supporting that sourcing is a key element for probiotic safety and or efficacy.

  April 25, 2017   doi: 10.1002/jcp.25814   open full text
• SUMO‐modified insulin‐like growth factor 1 receptor (IGF‐1R) increases cell cycle progression and cell proliferation.
  Yingbo Lin, Hongyu Liu, Ahmed Waraky, Felix Haglund, Prasoon Agarwal, Helena Jernberg‐Wiklund, Dudi Warsito, Olle Larsson.
  Journal of Cellular Physiology. April 25, 2017
  Increasing number of studies have shown nuclear localization of the insulin‐like growth factor 1 receptor (nIGF‐1R) in tumor cells and its links to adverse clinical outcome in various cancers. Any obvious cell physiological roles of nIGF‐1R have, however, still not been disclosed. Previously, we reported that IGF‐1R translocates to cell nucleus and modulates gene expression by binding to enhancers, provided that the receptor is SUMOylated. In this study, we constructed stable transfectants of wild type IGF1R (WT) and triple‐SUMO‐site‐mutated IGF1R (TSM) using igf1r knockout mouse fibroblasts (R‐). Cell clones (R‐WT and R‐TSM) expressing equal amounts of IGF‐1R were selected for experiments. Phosphorylation of IGF‐1R, Akt, and Erk upon IGF‐1 stimulation was equal in R‐WT and R‐TSM. WT was confirmed to enter nuclei. TSM did also undergo nuclear translocation, although to a lesser extent. This may be explained by that TSM heterodimerizes with insulin receptor, which is known to translocate to cell nuclei. R‐WT proliferated substantially faster than R‐TSM, which did not differ significantly from the empty vector control. Upon IGF‐1 stimulation G1‐S‐phase progression of R‐WT increased from 12 to 38%, compared to 13 to 20% of R‐TSM. The G1‐S progression of R‐WT correlated with increased expression of cyclin D1, A, and CDK2, as well as downregulation of p27. This suggests that SUMO‐IGF‐1R affects upstream mechanisms that control and coordinate expression of cell cycle regulators. Further studies to identify such SUMO‐IGF‐1R dependent mechanisms seem important. We constructed stable transfectants of wild type IGF1R (R‐WT) and triple‐SUMO‐site‐mutated IGF1R (R‐TSM) using igf1r knockout mouse fibroblasts (R‐). R‐WT proliferated substantially faster than R‐TSM, which did not differ significantly from the empty vector control. Upon IGF‐1 stimulation G1‐S‐phase progression of R‐WT increased significantly stronger comparing to R‐TSM, which was correlated with expression changes of cell cycle regulatory proteins.

  April 25, 2017   doi: 10.1002/jcp.25818   open full text
• Understanding the role of structural integrity and differential expression of integrin profiling to identify potential therapeutic targets in breast cancer.
  Vishal Das, Gazal Kalyan, Saugata Hazra, Mintu Pal.
  Journal of Cellular Physiology. April 25, 2017
  Breast cancer is found to be the most prevalent neoplasm in women worldwide. Despite the function of physically tethering cells to the matrix, transmembrane protein integrins are crucially involved in diverse cellular functions such as cell differentiation, proliferation, invasion, migration, and metastasis. Dysregulation of integrins and their interactions with the cells and their microenvironment can trigger several signaling cues that determine the cell fate decision. In this review, we spotlight all pre‐existing integrin molecules, their structure, molecular interactions motifs, and function through several cross talks with kinase receptors. We also discuss the role of these integrins as potential prognostic and therapeutic targets and also in the regulation of breast cancer cells differentiation. Understanding of integrin structure and their motifs for ligand interactions in this context will enable the development of new therapeutic approaches to sensitize the tumors and their microenvironment to conventional therapy and overall suppress their metastatic phenotype. In this review, we spotlight all pre‐existing integrin molecules, their structure, molecular interactions motifs, and function through several cross talks with kinase receptors. We also discuss the role of these integrins as potential prognostic and therapeutic targets and also in the regulation of breast cancer cells differentiation. Understanding of integrin structure and their motifs for ligand interactions in this context will enable the development of new therapeutic approaches to sensitize the tumors and their microenvironment to conventional therapy and overall suppress their metastatic phenotype.

  April 25, 2017   doi: 10.1002/jcp.25821   open full text
• NSI‐189, a small molecule with neurogenic properties, exerts behavioral, and neurostructural benefits in stroke rats.
  Naoki Tajiri, David M. Quach, Yuji Kaneko, Stephanie Wu, David Lee, Tina Lam, Ken L. Hayama, Thomas G. Hazel, Karl Johe, Michael C. Wu, Cesar V. Borlongan.
  Journal of Cellular Physiology. April 25, 2017
  Enhancing neurogenesis may be a powerful stroke therapy. Here, we tested in a rat model of ischemic stroke the beneficial effects of NSI‐189, an orally active, new molecular entity (mol. wt. 366) with enhanced neurogenic activity, and indicated as an anti‐depressant drug in a clinical trial (Fava et al., , Molecular Psychiatry, DOI: 10.1038/mp.2015.178) and being tested in a Phase 2 efficacy trial (ClinicalTrials.gov, , ClinicalTrials.gov Identifier: NCT02695472) for treatment of major depression. Oral administration of NSI‐189 in adult Sprague–Dawley rats starting at 6 hr after middle cerebral artery occlusion, and daily thereafter over the next 12 weeks resulted in significant amelioration of stroke‐induced motor and neurological deficits, which was maintained up to 24 weeks post‐stroke. Histopathological assessment of stroke brains from NSI‐189‐treated animals revealed significant increments in neurite outgrowth as evidenced by MAP2 immunoreactivity that was prominently detected in the hippocampus and partially in the cortex. These results suggest NSI‐189 actively stimulated remodeling of the stroke brain. Parallel in vitro studies further probed this remodeling process and demonstrated that oxygen glucose deprivation and reperfusion (OGD/R) initiated typical cell death processes, which were reversed by NSI‐189 treatment characterized by significant attenuation of OGD/R‐mediated hippocampal cell death and increased Ki67 and MAP2 expression, coupled with upregulation of neurogenic factors such as BDNF and SCF. These findings support the use of oral NSI‐189 as a therapeutic agent well beyond the initial 6‐hr time window to accelerate and enhance the overall functional improvement in the initial 6 months post stroke. Here, we show that enhancing neurogenesis may be a powerful stroke therapy. The present study tested in a rat model of ischemic stroke the beneficial effects of NSI‐189, an orally active, new molecular entity (mol. wt. 366) with enhanced neurogenic activity, and indicated as an anti‐depressant drug in a clinical trial (Fava et al., , Molecular Psychiatry, DOI: 10.1038/mp.2015.178) and being tested in a Phase 2 efficacy trial (ClinicalTrials.gov, , ClinicalTrials.gov Identifier: NCT02695472) for treatment of major depression. Behavioral recovery in NSI‐189‐treated stroke animals was accompanied by significant increments in neurite outgrowth as evidenced by MAP2 immunoreactivity that was prominently detected in the hippocampus and partially in the cortex, indicating that NSI‐189 actively stimulated remodeling of the stroke brain.

  April 25, 2017   doi: 10.1002/jcp.25847   open full text
• Osteoactivin regulates head and neck squamous cell carcinoma invasion by modulating matrix metalloproteases.
  Oneida A. Arosarena, Eric W. Barr, Ryan Thorpe, Hilary Yankey, Joseph T. Tarr, Fayez F. Safadi.
  Journal of Cellular Physiology. April 25, 2017
  Nearly 60% of patients with head and neck squamous cell carcinoma (HNSCC) die of metastases or locoregional recurrence. Metastasis is mediated by cancer cell migration and invasion, which are in part dependent on extracellular matrix degradation by matrix metalloproteinases. Osteoactivin (OA) overexpression plays a role in metastases in several malignancies, and has been shown to upregulate matrix metalloproteinase (MMP) expression and activity. To determine how OA modulates MMP expression and activity in HNSCC, and to investigate OA effects on cell invasion, we assessed effects of OA treatment on MMP mRNA and protein expression, as well as gelatinase and caseinolytic activity in HNSCC cell lines. We assessed the effects of OA gene silencing on MMP expression, gelatinase and caseinolytic activity, and cell invasion. OA treatment had differential effects on MMP mRNA expression. OA treatment upregulated MMP‐10 expression in UMSCC14a (p = 0.0431) and SCC15 (p < 0.0001) cells, but decreased MMP‐9 expression in UMSCC14a cells (p = 0.0002). OA gene silencing decreased MMP‐10 expression in UMSCC12 cells (p = 0.0001), and MMP‐3 (p = 0.0005) and ‐9 (p = 0.0036) expression in SCC25 cells. In SCC15 and SCC25 cells, OA treatment increased MMP‐2 (p = 0.0408) and MMP‐9 gelatinase activity (p < 0.0001), respectively. OA depletion decreased MMP‐2 (p = 0.0023) and ‐9 (p < 0.0001) activity in SCC25 cells. OA treatment increased 70 kDa caseinolytic activity in UMSCC12 cells consistent with tissue type plasminogen activator (p = 0.0078). OA depletion decreased invasive capacity of UMSCC12 cells (p < 0.0001). OA's effects on MMP expression in HNSCC are variable, and may promote cancer cell invasion. Osteoactivin/gpnmb has been implicated in metastases, particularly bony metastases, in several malignancies. In this study we demonstrated that osteoactivin differentially upregulates matrix metalloprotease expression and activity in head and neck cancer cell lines.

  April 25, 2017   doi: 10.1002/jcp.25900   open full text
• High glucose alters the secretome of mechanically stimulated osteocyte‐like cells affecting osteoclast precursor recruitment and differentiation.
  Marta Maycas, María Teresa Portolés, María Concepción Matesanz, Irene Buendía, Javier Linares, María José Feito, Daniel Arcos, María Vallet‐Regí, Lilian I. Plotkin, Pedro Esbrit, Arancha R. Gortázar.
  Journal of Cellular Physiology. April 25, 2017
  Diabetes mellitus (DM) induces bone deterioration, while mechanical stimulation promotes osteocyte‐driven bone formation. We aimed to evaluate the interaction of acute exposure (24 h) to high glucose (HG) with both the pro‐survival effect conferred to osteocytic MLO‐Y4 cells and osteoblastic MC3T3‐E1 cells by mechanical stimulation and the interaction of these cells with osteoclast precursor RAW264.7 cells. We found that 24 h of HG (25 mM) pre‐exposure prevented both cell survival and ERK and β‐catenin nuclear translocation upon mechanical stimulation by fluid flow (FF) (10 min) in both MLO‐Y4 and MC3T3‐E1 cells. However, migration of RAW 264.7 cells was inhibited by MLO‐Y4 cell‐conditioned medium (CM), but not by MC3T3‐E1 cell‐CM, with HG or FF. This inhibitory effect was associated with consistent changes in VEGF, RANTES, MIP‐1α, MIP‐1β MCP‐1, and GM‐CSF in MLO‐Y4 cell‐CM. RAW264.7 proliferation was inhibited by MLO‐Y4 CM under static or HG conditions, but it increased by FF‐CM with or without HG. In addition, both FF and HG abrogated the capacity of RAW 264.7 cells to differentiate into osteoclasts, but in a different manner. Thus, HG‐CM in static condition allowed formation of osteoclast‐like cells, which were unable to resorb hydroxyapatite. In contrast, FF‐CM prevented osteoclastogenesis even in HG condition. Moreover, HG did not affect basal RANKL or IL‐6 secretion or their inhibition induced by FF in MLO‐Y4 cells. In conclusion, this in vitro study demonstrates that HG exerts disparate effects on osteocyte mechanotransduction, and provides a novel mechanism by which DM disturbs skeletal metabolism through altered osteocyte‐osteoclast communication. This in vitro study demonstrates that HG exerts disparate effects on osteocyte mechanotransduction, and provides a novel mechanism by which DM disturbs skeletal metabolism through altered osteocyte‐osteoclast communication.

  April 25, 2017   doi: 10.1002/jcp.25829   open full text
• Diagnostic and Therapeutic Potential of Exosomes in Cancer: The Beginning of a New Tale?
  Hamed Mirzaei, Amirhossein Sahebkar, Mahmoud Reza Jaafari, Mohammad Goodarzi, Hamid Reza Mirzaei.
  Journal of Cellular Physiology. April 25, 2017
  Exosomes have emerged as one of the main players in intercellular communication. These small nano‐sized particles have many roles in various physiological pathways in normal and abnormal cells. Exosomes can carry various cargos such as proteins, mRNAs, and miRNAs to recipient cells. Uptake of exosomes and their cargo can induce and/or inhibit different cellular and molecular pathways that lead to the alteration of cell behavior. Multiple lines of evidence have indicated that exosomes released from cancer cells can effect development of cancer in different stages. These particles and their cargo could regulate different processes such as tumor growth, metastasis, drug resistance, angiogenesis, and immune system functioning. It has been observed that exosomes can be used as potential diagnostic biomarkers in various cancer types. Moreover, some studies have used these particles as biological vehicles for delivery of various drugs such as doxorubicin, siRNAs, and miRNAs. Here, we summarized the findings on the role of exosomes in different pathological processes involved in cancer. Moreover, application of these particles as diagnostic and therapeutic biomarkers in different types of cancers is discussed. J. Cell. Physiol. 9999: 1–10, 2016. © 2016 Wiley Periodicals, Inc.

  April 25, 2017   doi: 10.1002/jcp.25739   open full text
• sp2‐Iminosugar α‐glucosidase inhibitor 1‐C‐octyl‐2‐oxa‐3‐oxocastanospermine specifically affected breast cancer cell migration through Stim1, β1‐integrin, and FAK signaling pathways.
  Nahla Gueder, Ghada Allan, Marie‐Sophie Telliez, Frédéric Hague, José M. Fernandez, Elena M. Sanchez‐Fernandez, Carmen Ortiz‐Mellet, Ahmed Ahidouch, Halima Ouadid‐Ahidouch.
  Journal of Cellular Physiology. April 25, 2017
  Aberrant glycosylation changes on many glycoproteins are often related to cancer progression and metastasis. sp2‐Iminosugar‐type castanospermine analogues, inhibitors of α‐glucosidases, have been reported to exhibit antitumor activity. However, their effects on cell migration and the underlying molecular mechanism are not fully understood. Here, we investigated the effect of the pseudo‐C‐octyl glycoside 2‐oxa‐3‐oxocastanospermine derivatives (CO‐OCS) on breast cancer cells (MCF‐7 and MDA‐MB‐231 cells), and MCF‐10A mammary normal cell lines. We showed that CO‐OCS treatment results in the drastic decrease of breast cancer cell migration without affecting cell proliferation. Furthermore, CO‐OCS significantly reduced both the expression of β1‐integrin, which is a crucial interacting partner of Focal Adhesion Kinase (FAK), and the phosphorylation rates of FAK and ERK1/2. CO‐OCS also drastically reduced Ca2+ entry through Store Operated Channels (SOC). Orai1 and Stim1, two N‐glycosylated proteins, are involved in Store‐Operated Calcium Entry (SOCE), and are essential for breast tumor cell migration. Our results showed that CO‐OCS decreased the expression, at the protein level, of Stim1 without affecting that of Orai1. Moreover, cell migration and SOCE were attenuated by CO‐OCS as well as when Stim1 was silenced. In contrast, in MCF‐10A cells, CO‐OCS slightly reduced cell migration, but was without effect on gene expression of Stim1, Orai1, β1‐integrin, or FAK and ERK1/2 activation. Our results provide strong evidence for a significant effect of CO‐OCS on breast cancer cell migration and support that this effect was associated with β1‐integrin, Stim1, and FAK signaling pathways. CO‐OCS suppressed more drastically migration of breast cancer cells than normal cells CO‐OCS affected specifically breast cancer cell migration by targeting β1‐integrin and Stim1 CO‐OCS does not affect the expression of β1‐integrin, or Stim1 in MCF‐10A normal cells.

  April 25, 2017   doi: 10.1002/jcp.25832   open full text
• Up‐Regulation of Transient Receptor Potential Melastatin 6 Channel Expression by Tumor Necrosis Factor‐α in the Presence of Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor.
  Chisa Furukawa, Naoko Fujii, Aya Manabe, Toshiyuki Matsunaga, Satoshi Endo, Hajime Hasegawa, Yoshinori Ito, Masahiko Yamaguchi, Yasuhiro Yamazaki, Akira Ikari.
  Journal of Cellular Physiology. April 25, 2017
  Anti‐epidermal growth factor receptor (EGFR) drugs such as erlotinib and gefitinib cause a side effect of hypomagnesemia, but chemotherapy to treat this has not yet been developed. The transient receptor potential melastatin 6 (TRPM6) channel is involved in the reabsorption of Mg2+ in the renal tubule. We reported previously that the expression of TRPM6 is up‐regulated by epidermal growth factor (EGF) in renal tubular epithelial NRK‐52E and HEK293 cells. EGF‐induced elevation of TRPM6 expression was inhibited by erlotinib, gefitinib, and lapatinib. We found that tumor necrosis factor‐α (TNF‐α) increases TRPM6 expression in the presence of erlotinib. Therefore, we investigated what molecules are involved in the up‐regulation of TRPM6 expression by TNF‐α. EGF increased the levels of phosphorylated extracellular signal‐regulated kinase 1 and 2 (p‐ERK1/2), which were inhibited by erlotinib. TNF‐α did not change p‐ERK1/2 levels, but increased the phosphorylation and nuclear localization of nuclear factor‐κB (NF‐κB), which were blocked by the NF‐κB inhibitors BAY 11–7082 and pyrrolidinedithiocarbamate ammonium. Similarly, luciferase reporter activity of human TRPM6 was increased by TNF‐α, which was blocked by NF‐κB inhibitors, and was inhibited by a mutation in the κB‐binding site in the proximal region of the TRPM6 promoter. A chromatin immunoprecipitation assay revealed that NF‐κB binds to the κB‐binding site, which was blocked by NF‐κB inhibitors. In the presence of erlotinib, TNF‐α increased Mg2+ influx, which was blocked by NF‐κB inhibitors. These results suggest that TNF‐α reverses the reduction in Mg2+ reabsorption caused by anti‐EGFR drugs. J. Cell. Physiol. 232: 2841–2850, 2017. © 2016 Wiley Periodicals, Inc. Tumor necrosis factor‐α (TNF‐α) increases TRPM6 expression in the presence of anti‐epidermal growth factor receptor drug, erlotinib. TNF‐α did not change p‐ERκ1/2 levels, but increased the phosphorylation and nuclear localization of nuclear factor‐κB (NF‐κB), which were blocked by the NF‐κB inhibitors. In the presence of erlotinib, TNF‐α increased Mg2+ influx, which was blocked by NF‐κB inhibitors.

  April 25, 2017   doi: 10.1002/jcp.25709   open full text
• Brain‐derived neurotrophic factor improves proliferation of endometrial epithelial cells by inhibition of endoplasmic reticulum stress during early pregnancy.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. April 25, 2017
  Brain‐derived neurotrophic factor (BDNF) is a member of the neurotrophin family binds to two transmembrane receptors; neurotrophic receptor tyrosine kinase 2 (NTRK2) with high affinity and p75 with low affinity. Although BDNF‐NTRK2 signaling in the central nervous system is known, signaling in the female reproductive system is unknown. Therefore, we determined effects of BDNF on porcine endometrial luminal epithelial (pLE) cells isolated from Day 12 of pregnancy, as well as expression of BDNF and NTRK2 in endometria of cyclic and pregnant pigs. BDNF‐NTRK2 genes were expressed in uterine glandular (GE) and luminal (LE) epithelia during early pregnancy. In addition, their expression in uterine GE and LE decreased with increasing parity of sows. Recombinant BDNF increased proliferation in pLE cells in a dose‐dependent, as well as expression of PCNA and Cyclin D1 in nuclei of pLE cells. BDNF also activated phosphorylation of AKT, P70S6K, S6, ERK1/2, JNK, P38 proteins in pLE cells. In addition, cell death resulting from tunicamycin‐induced ER stress was prevented when pLE cells were treated with the combination of tunicamycin and BDNF which also decreased cells in the Sub‐G1 phase of the cell cycle. Furthermore, tunicamycin‐induced unfolded protein response genes were mostly down‐regulated to the basal levels as compared to non‐treated pLE cells. Our finding suggests that BDNF acts via NTRK2 to induce development of pLE cells for maintenance of implantation and pregnancy by activating cell signaling via the PI3K and MAPK pathways and by inhibiting ER stress. BDNF acts via NTRK2 to induce development of the uterine epithelial cells for maintenance of implantation and pregnancy by activating cell signaling via the PI3K and MAPK pathways and by inhibiting ER stress during early pregnancy.

  April 25, 2017   doi: 10.1002/jcp.25834   open full text
• WNT4 Acts Downstream of BMP2 to Mediate the Regulation of ATRA Signaling on RUNX1 Expression: Implications for Terminal Differentiation of Antler Chondrocytes.
  Hong‐Liang Zhang, Zhan‐Qing Yang, Cui‐Cui Duan, Shuang Geng, Kai Wang, Hai‐Fan Yu, Zhan‐Peng Yue, Bin Guo.
  Journal of Cellular Physiology. April 24, 2017
  Although ATRA is involved in regulating the proliferation and differentiation of chondrocytes, its underlying mechanism remains unknown. Here we showed that ATRA could stimulate the proliferation of antler chondrocytes and expression of COL X and MMP13 which were two well‐known markers for hypertrophic chondrocytes. Silencing of CRABP2 prevented the induction of ATRA on chondrocyte terminal differentiation, while overexpression of CRABP2 exhibited the opposite effects. CYP26A1 and CYP26B1 weakened the sensitivity of antler chondrocytes to ATRA. Further analysis evidenced that ATRA might induce chondrocyte terminal differentiation and modulate the expression of BMP2, WNT4 and RUNX1 through RARα/RXRα. Knockdown of BMP2 enhanced the induction of ATRA on the expression of COL X and MMP13, whereas overexpression of BMP2 abrogated this effectiveness. WNT4 might mediate the effects of ATRA and BMP2 on chondrocyte terminal differentiation. Dysregulation of BMP2 impaired the regulation of ATRA on WNT4 expression. Administration of ATRA to antler chondrocytes transfected with RUNX1 siRNA failed to induce the differentiation. Conversely, rRUNX1 strengthened the stimulation of ATRA on the expression of COL X and MMP13. Simultaneously, RUNX1 was a downstream effector of BMP2 and WNT4 in chondrocyte terminal differentiation. Moreover, WNT4 might play an important role in the crosstalk between BMP2 and RUNX1. Attenuation of BMP2 or WNT4 enhanced the interaction between ATRA and RUNX1, while constitutive expression of BMP2 or WNT4 reversed the regulation of ATRA on RUNX1. Collectively, WNT4 may act downstream of BMP2 to mediate the effects of ATRA on the terminal differentiation of antler chondrocytes through targeting RUNX1. This article is protected by copyright. All rights reserved

  April 24, 2017   doi: 10.1002/jcp.25972   open full text
• Generation of Insulin‐Producing Cells From Human‐Induced Pluripotent Stem Cells Using a Stepwise Differentiation Protocol Optimized With Platelet‐Rich Plasma.
  Seyed Ehsan Enderami, Yousef Mortazavi, Masoud Soleimani, Samad Nadri, Alireza Biglari, Reyhaneh Nassiri Mansour.
  Journal of Cellular Physiology. April 21, 2017
  Studies on patient‐specific human‐induced pluripotent stem cells (hiPSCs) as well as a series of autologous growth factors presumably will reveal their many benefits for cell base replacement therapy in type 1 diabetes mellitus (TIDM) patients in the future. For this purpose, we established a multistep protocol by adding platelet‐rich plasma (PRP) that induce the hiPSCs into insulin‐producing cells (IPCs). We present here a differentiation protocol consisting of five stages in two groups including protocol with PRP and without PRP. Charac­teristics of derived IPCs in both groups were evaluated at the mRNA and protein levels, cell cycle and viability in the end stage of cell differentiation. The in vitro studies indicated the treatment of hiPSCs in the protocol with PRP resulting in differentiated cells with strong characteristics of IPCs including islet‐like cells, the expression of mature and functional pancreatic beta cell specific marker genes. In addition to these pancreatic specific markers were detected by immunochemistry and Western blot. Our differentiated cells in two groups secreted insulin and C‐peptide in a glucose stimulation test by ELISA showing in vitro functional. The results of the cell cycle assay confirmed that differentiation has been done. We reported for the first time that PRP might be ideal additive in the culture medium to induce pancreatic differentiation in the hiPSCs. This study provides a new approach to investigate the role of PRP in pancreatic differentiation protocols and enhance the feasibility of using patient‐specific iPSCs and autologous PRP for future beta cells replacement therapies for T1DM. J. Cell. Physiol. 232: 2878–2886, 2017. © 2016 Wiley Periodicals, Inc. We established a multistep protocol by adding platelet‐rich plasma (PRP) that induce the hiPSCs into insulin‐producing cells (IPCs). We reported for the first time that PRP might be ideal additive in the culture medium to induce pancreatic differentiation in the hiPSCs.

  April 21, 2017   doi: 10.1002/jcp.25721   open full text
• Purification of small molecule‐induced cardiomyocytes from human induced pluripotent stem cells using a reporter system.
  Geun Hye Hwang, So Mi Park, Ho Jae Han, Joong Sun Kim, Seung Pil Yun, Jung Min Ryu, Ho Jin Lee, Woochul Chang, Su‐Jin Lee, Jeong‐Hee Choi, Jin‐Sung Choi, Min Young Lee.
  Journal of Cellular Physiology. April 18, 2017
  In order to realize the practical use of human pluripotent stem cell (hPSC)‐derived cardiomyocytes for the purpose of clinical use or cardiovascular research, the generation of large numbers of highly purified cardiomyocytes should be achieved. Here, we show an efficient method for cardiac differentiation of human induced pluripotent stem cells (hiPSCs) in chemically defined conditions and purification of hiPSC‐derived cardiomyocytes using a reporter system. Regulation of the Wnt/β‐catenin signaling pathway is implicated in the induction of the cardiac differentiation of hPSCs. We increased cardiac differentiation efficiency of hiPSCs in chemically defined conditions through combined treatment with XAV939, a tankyrase inhibitor and IWP2, a porcupine inhibitor and optimized concentrations. Although cardiac differentiation efficiency was high (>80%), it was difficult to suppress differentiation into non‐cardiac cells, Therefore, we applied a lentiviral reporter system, wherein green fluorescence protein (GFP) and Zeocin‐resistant gene are driven by promoter activation of a gene (TNNT2) encoding cardiac troponin T (cTnT), a cardiac‐specific protein, to exclude non‐cardiomyocytes from differentiated cell populations. We transduced this reporter construct into differentiated cells using a lentiviral vector and then obtained highly purified hiPSC‐derived cardiomyocytes by treatment with the lowest effective dose of Zeocin. We significantly increased transgenic efficiency through manipulation of the cells in which the differentiated cells were simultaneously infected with virus and re‐plated after single‐cell dissociation. Purified cells specifically expressed GFP, cTnT, displayed typical properties of cardiomyocytes. This study provides an efficient strategy for obtaining large quantities of highly purified hPSC‐derived cardiomyocytes for application in regenerative medicine and biomedical research. Combined treatment with XAV939 and IWP2 enhances hiPSC cardiac differentiation. Large amount of highly purified hiPSC‐derived cardiomyocytes are obtained using a reporter system. Purified cells represent functional cardiomyocytes that possess electrophysiological properties and form cardiac junctions.

  April 18, 2017   doi: 10.1002/jcp.25783   open full text
• Establishment of a new conditionally immortalized human skeletal muscle microvascular endothelial cell line.
  Hironori Sano, Yasuteru Sano, Eri Ishiguchi, Fumitaka Shimizu, Masatoshi Omoto, Toshihiko Maeda, Hideaki Nishihara, Yukio Takeshita, Shiori Takahashi, Mariko Oishi, Takashi Kanda.
  Journal of Cellular Physiology. April 18, 2017
  In skeletal muscle, the capillaries have tight junctions (TJs) that are structurally similar to those in the blood‐brain barrier (BBB) and blood‐nerve barrier (BNB). Although many findings have been clarified in the territory of BBB and BNB, few have so far examined the TJs of capillaries in the skeletal muscle. In addition, no in vitro human skeletal muscle microvasculature models have been reported thus far. We newly established a new human skeletal muscle microvascular endothelial cell (HSMMEC) line. HSMMECs were isolated from human skeletal muscle and were infected with retroviruses harboring temperature‐sensitive SV40 T antigen and telomerase genes. This cell line, termed TSM15, showed a spindle fiber‐shaped morphology, an immunoreactivity to anti‐factor VIII and anti‐VE‐cadherin antibodies, and a temperature‐sensitive growth. TSM15 cells grew stably for more than 40 passages when they were cultured at 33°C, thereby retaining their spindle fiber‐shaped morphology and contact inhibition at confluence. The cells expressed tight junctional molecules such as claudin‐5, occludin, and zonula occludens‐1, as well as transporters such as a glucose transporter 1. The transendothelial electrical resistance of TSM15 was as high as those of the human brain microvascular endothelial cell line. This novel cell line might facilitate the analyses of the pathophysiology of inflammatory myopathy, such as dermatomyositis, and can improve our understanding of the physiological and biochemical properties of the microvasculature in human skeletal muscle. A new conditionally immortalized human skeletal muscle microvascular endothelial cell line was established. This cell line, termed TSM15, expressed tight junctional molecules as well as transporters and has a barrier integrity relevant to that of a human brain microvascular endothelial cell line.

  April 18, 2017   doi: 10.1002/jcp.25772   open full text
• GM13133 is a negative regulator in mouse white adipocytes differentiation and drives the characteristics of brown adipocytes.
  LiangHui You, YaHui Zhou, XianWei Cui, XingYun Wang, YaZhou Sun, Yao Gao, Xing Wang, Juan Wen, Kaipeng Xie, RanRan Tang, ChenBo Ji, XiRong Guo.
  Journal of Cellular Physiology. April 18, 2017
  Obesity is tightly associated with the disturbance of white adipose tissue storing excess energy. Thermogenic adipocytes (brown and beige) exert a critical role of oxidizing nutrients at the high rates through non‐shivering thermogenesis. The recruitment of brown characteristics in white adipocytes, termed browning, has been considered as a promising strategy for treating obesity and associated metabolic complications. Recently, long noncoding RNAs play a crucial role in regulating tissue development and participating in disease pathogenesis, yet their effects on the conversion of white into brown‐like adipocytes and thermogenic function were not totally understood. Here, we identified a mouse brown adipose specific expressed lncRNA, termed GM13133. Moreover, a considerable amount of GM13133 is expressed in adipocytes and actively modulated by cold, β3‐adrenergic agonist and cAMP stimuli, implying a potential role in the conversion from white to brown adipocytes. Overexpression of GM13133 did not affect the proliferation of mouse white pre‐adipocytes, but inhibited white adipocyte differentiation by decreasing lipid accumulation. The forced expression of GM13133 also significantly drove the conversion of white into brown‐like adipocytes with the enhanced mitochondrial biogenesis and the induced expression of brown adipocytes specific markers. A global mRNA analysis further indicated the possible regulatory role of cAMP signaling pathway in GM13133 mediated white‐to‐brown adipocytes conversion. Our results identified a lncRNA‐mediated modulation in primary mouse white adipocyte differentiation and indicate the functional significance of GM13133 in promoting browning of white adipocytes and maintenance of thermogenesis, further providing a potential strategy to treating obesity. Our results provide identified a lncRNA‐mediated modulation in primary mouse white adipocyte differentiation and further indicate the functional significance of GM13133 in promoting browning of white adipocytes and maintenance of thermogenesis, further providing a potential strategy to treating obesity.

  April 18, 2017   doi: 10.1002/jcp.25878   open full text
• Evaluation the ethno‐pharmacological studies in Iran during 2004–2016: A systematic review.
  Zahra Sadeghi, Maryam Akaberi, Alireza Sobhkhizi, Amirhossein Sahebkar, Seyed Ahmad Emami.
  Journal of Cellular Physiology. April 18, 2017
  Although Iran has a deep history in herbal medicine and great heritage of ancient medical scholars, few efforts have been made to evaluate ethnopharmacological aspects of medicinal plants in this country. In the present study, the authors have reviewed all important literature about the ethnopharmacological investigations on medicinal plants used in the last decade in Iran. All provinces of Iran were categorized according to a phytogeographical division. Information was collected through bibliographic investigations from scientific journals and books. Afterward, the data were analyzed through the construction of specific ecological regions of the country. Fifty‐five references reporting medicinal plants in five ecological zones were retrieved. The Irano‐Turanian subregion has produced the greatest number of publications in this field among others (47%). Results illustrate that the most reported botanical families were Lamiaceae and Asteraceae (28.57% and 27.73%, respectively). Among various illnesses reported for these plants, gastrointestinal (30.15%), respiratory problems (14.28%), diabetes (11.11%), and cold/flu (11.11%) were the most cited. The most frequently cited medicinal uses were attributed to decoction and infusion preparations. Iran has a rich history of knowledge about phytotherapy and has also a diverse geographical regions, and a plant flora that is a good candidate for drug discovery. Documentation of indigenous knowledge about herbal medicine used by Iranian tribes is vital for the future development of herbal drugs. Ethnopharmacological studies of Iranian folk medicine with quantitative analytical techniques are warranted to find drug candidates, and also to preserve the precious knowledge of the Iranian folk medicine. This is the first ethno‐pharmacological survery of medicinal plants used in the treatment of different diseases in Iran.

  April 18, 2017   doi: 10.1002/jcp.25803   open full text
• Osteogenic differentiation Potential of Mesenchymal Stem Cells cultured on Nanofibrous Scaffold Improved in the Presence of Pulsed Electromagnetic Field.
  Monireh Arjmand, Abdolreza Ardeshirylajimi, Hossein Maghsoudi, Esmaeel Azadian.
  Journal of Cellular Physiology. April 17, 2017
  Nowadays, tissue engineering by using stem cells in combination with scaffolds and bioactive molecules has made significant contributions to the regeneration of damaged bone tissues. Since the usage of bioactive molecules including, growth factors to induce differentiation is safety limited in clinical applications, and it has also been previously observed that extremely low frequency pulsed electromagnetic fields (PEMF) can be effective in the enhancement of proliferation rate and osteogenic differentiation of stem cells, the aim of this study was investigating the osteoinductive potential of PEMF in combination with Poly(caprolactone) (PCL) nanofibrous scaffold. To achieve this aim, Adipose‐derived mesenchymal stem cells (ADSCs) isolated and characterized and then osteogenic differentiation of them was investigated after culturing on the surface of PCL scaffold under treatments of PEMF, PEMF plus osteogenic medium (OM) and OM. Analysis of common osteogenic markers such as Alizarin red staining, ALP activity, calcium content and four important bone‐related genes in days of 7, 14 and 21 confirmed that the effects of PEMF on the osteogenic differentiation of ADSCs are very similar to the effects of osteogenic medium. Thus, regarding the immunological concerns about the application of bioactive molecules for tissue engineering, PEMF could be a good alternative for osteogenic medium. Although, results were showed a synergetic effect for simultaneous application of PEMF and PCL scaffold in the osteogenesis process of ADSCs. Taking together, ADSCs‐seeded PCL nanofibrous scaffold in combination with PEMF could be a great option for use in bone tissue engineering applications. This article is protected by copyright. All rights reserved

  April 17, 2017   doi: 10.1002/jcp.25962   open full text
• Activation of ClC‐3 chloride channel by 17β‐estradiol relies on the estrogen receptor α expression in breast cancer.
  Haifeng Yang, Lianshun Ma, Yawei Wang, Wanhong Zuo, Bingxue Li, Yaping Yang, Yehui Chen, Lixin Chen, Liwei Wang, Linyan Zhu.
  Journal of Cellular Physiology. April 17, 2017
  Although extensively studied, the mechanisms by which estrogen promotes breast cancer growth remain to be fully elucidated. Tamoxifen, an antiestrogen to treat ERα+ breast cancer, is also a high‐affinity blocker of the chloride channels. In this study, we explored the involvement of the chloride channels in the action of estrogen in breast cancer. We found that 17β‐estradiol (17β‐E2) concentration‐dependently activated the chloride currents in ERα+ breast cancer MCF‐7 cells. Extracellular hypertonic challenge and chloride channel blockers, NPPB and DIDS inhibited the 17β‐E2‐activated chloride currents. Decreased the ClC‐3 protein expression caused the depletion of the 17β‐E2‐activated chloride currents. 17β‐E2‐activated chloride currents which relied on the ERα expression were demonstrated by the following evidences. Firstly, 17β‐E2‐activated chloride currents could not be observed in ERα‐ breast cancer MDA‐MB‐231 cells. Secondly, ER antagonists, tamoxifen and ICI 182,780, and downregulation of ERα expression inhibited or abolished the 17β‐E2‐activated chloride currents. Thirdly, ERα expression was induced in MDA‐MB‐231 cells by ESR1 gene transfection, and then 17β‐E2‐activated chloride currents could be observed. In MCF‐7 cells, ERα and ClC‐3 mainly located in nucleus and translocated to cell plasma and membrane with respect to co‐localization following treatment of 17β‐E2. Downregulation of ERα expression could decrease the expression of ClC‐3 protein. Conversely, downregulation of ClC‐3 expression did not influence the ERα expression. Taken together, our findings demonstrated that ClC‐3 is a potential target of 17β‐E2 and modulates by the ERα in breast cancer cell. Pharmacological modulation of ClC‐3 may provide a deep understanding in antiestrogen treatment of breast cancer patients. This article is protected by copyright. All rights reserved

  April 17, 2017   doi: 10.1002/jcp.25963   open full text
• Platelet‐derived microparticles regulates thrombin generation via phophatidylserine in abdominal sepsis.
  Yongzhi Wang, Su Zhang, Lingtao Luo, Eva Norström, Oscar Ö Braun, Matthias Mörgelin, Henrik Thorlacius.
  Journal of Cellular Physiology. April 14, 2017
  Sepsis is associated with dysfunctional coagulation. Recent data suggest that platelets play a role in sepsis by promoting neutrophil accumulation. Herein, we show that cecal ligation and puncture (CLP) triggered systemic inflammation, which is characterized by formation of IL‐6 and CXC chemokines as well as neutrophil accumulation in the lung. Platelet depletion decreased neutrophil accumulation, IL‐6 and CXC chemokines formation in septic lungs. Depletion of platelets increased peak thrombin formation and total thrombin generation (TG) in plasma from septic animals. CLP elevated circulating levels of platelet‐derived microparticles (PMPs). In vitro generated PMPs were a potent inducer of TG. Interestingly, in vitro wild‐type recombinant annexin V abolished PMP‐induced thrombin formation whereas a mutant annexin V protein, which does not bind to phosphatidylserine (PS), had no effect. Administration of wild‐type, but not mutant annexin V, significantly inhibited thrombin formation in septic animals. Moreover, CLP‐induced formation of thrombin‐antithrombin complexes were reduced in platelet‐depleted mice and in animals pretreated with annexin V. PMP‐induced TG attenuated in FXII‐ and FVII‐deficient plasma. These findings suggest that sepsis‐induced TG is dependent on platelets. Moreover, PMPs formed in sepsis are a potent inducer of TG via PS exposure and activation of both the intrinsic and extrinsic pathway of coagulation. In conclusion, these observations suggest that PMPs and PS play an important role in dysfunctional coagulation in abdominal sepsis. This article is protected by copyright. All rights reserved

  April 14, 2017   doi: 10.1002/jcp.25959   open full text
• Spermicidal efficacy of VRP, a synthetic cationic antimicrobial peptide, inducing apoptosis and membrane disruption.
  Prasanta Ghosh, Arpita Bhoumik, Sudipta Saha, Sandipan Mukherjee, Sarfuddin Azmi, Jimut Kanti Ghosh, Sandhya Rekha Dungdung.
  Journal of Cellular Physiology. April 14, 2017
  Presently available contraceptives are mostly hormonal or detergent in nature with numerous side effects like irritation, lesion, inflammation in vagina, alteration of body homeostasis etc. Antimicrobial peptides with spermicidal activity but without adverse effects may be suitable alternatives. In the present study spermicidal activity of a cationic antimicrobial peptide VRP on human spermatozoa has been elucidated. Progressive forward motility of human spermatozoa was instantly stopped after 100 µM VRP treatment and at 350 µM all kinds of sperm motility ceased within 20 seconds as assessed by the Sander‐Cramer assay. The spermicidal effect was confirmed by eosin‐nigrosin assay and HOS test. VRP treatment (100µM) in human spermatozoa induced both the intrinsic and extrinsic pathways of apoptosis. TUNEL assay showed VRP treatment significantly disrupted the DNA integrity and changed the mitochondrial membrane permeability as evident from MPTP assay. AFM and SEM results depicted ultra structural changes including disruption of the acrosomal cap and plasma membrane of the head and midpiece region after treatment with 350 µM VRP. MTT assay showed after treatments with 100 µM and 350 µM of VRP for 24 hrs, a substantial amount of Lactobacillus acidophilus (about 90% and 75% respectively) remained viable. Hence, VRP being a small synthetic peptide with antimicrobial and spermicidal activity but tolerable to normal vaginal microflora, may be a suitable target for elucidating its contraceptive potentiality. This article is protected by copyright. All rights reserved

  April 14, 2017   doi: 10.1002/jcp.25958   open full text
• Metformin attenuates albumin‐induced alterations in renal tubular cells in vitro.
  Soumaya Allouch, Shankar Munusamy.
  Journal of Cellular Physiology. April 13, 2017
  Proteinuria (albuminuria) plays a crucial role in the etiology of chronic kidney disease (CKD) via alteration of multiple signaling pathways and cellular process in renal cells. The objectives of this study are to investigate the effects of activation of the energy‐sensing molecule AMP‐activated kinase (AMPK) in renal cells using metformin on endoplasmic reticulum (ER) stress, AKT, mTOR, epithelial‐to‐mesenchymal transition (EMT), autophagy, and apoptosis that are thought to mediate renal cell injury during proteinuria, and to dissect the AMPK‐ and non‐AMPK mediated effects of metformin using an in vitro model of albumin‐induced renal cell injury. Rat renal proximal tubular (NRK‐52E) cells were exposed to 10 and 15 mg/ml of albumin for 72 h in the presence of 1 mM Metformin and/or 0.5 µM compound C, and assessed for alterations in the aforementioned pathways. Metformin treatment restored AMPK phosphorylation and augmented autophagy in renal cells exposed to albumin. In addition, metformin treatment attenuated the albumin‐induced phosphorylation of AKT and the downstream targets of mTOR, and prevented albumin‐mediated inductions of EMT marker (α‐SMA), pro‐apoptotic ER stress marker CHOP, and apoptotic caspases ‐12 and ‐3 in renal cells. Blockade of metformin‐induced AMPK activation with compound C blunted the ER defense response and autophagy but had no effect on the markers of EMT and apoptosis in our model. Our studies suggest that metformin protects renal cells against proteinuric cytotoxicity via suppression of AKT and mTOR activation, inhibition of EMT and apoptosis, and augmentation of autophagy and ER defense response through AMPK‐independent and AMPK‐dependent mechanisms, respectively. Proteinuria (albuminuria) plays a crucial role in the etiology of chronic kidney disease via alteration of multiple signaling pathways in renal cells. The current study using an in vitro model of albumin‐induced cytotoxicity suggests that metformin protects renal cells against proteinuric cytotoxicity via suppression of AKT and mTOR activation, inhibition of epithelial‐to‐mesenchymal transition (EMT) and apoptosis, and augmentation of autophagy and ER defense response through AMPK‐independent and AMPK‐dependent mechanisms respectively.

  April 13, 2017   doi: 10.1002/jcp.25838   open full text
• Anti‐tumor effects of crocetin and related molecular targets.
  Maliheh Moradzadeh, Hamid Reza Sadeghnia, Alijan Tabarraei, Amirhossein Sahebkar.
  Journal of Cellular Physiology. April 13, 2017
  Natural products have gained a wide popularity as chemopreventive and anti‐cancer agents owing to their multi‐mechanistic mode of action, availability and synergism with several conventional chemotherapeutic agents. Crocetin is a carotenoid compound isolated from the stigma of Crocus sativus L. (saffron). Crocetin has shown promising effects as an anti‐tumor agent in animal models and cell culture systems. Crocetin retards the growth of cancer cells via inhibiting nucleic acid synthesis, enhancing anti‐oxidative system, and inducing apoptosis and differentiation pathways. The present review outlines natural sources of crocetin, and its pharmacokinetic and pharmacological properties relevant to the prevention and treatment of cancer. Also, we discuss molecular targets underlying the putative anti‐tumor effects of crocetin. This article is protected by copyright. All rights reserved

  April 13, 2017   doi: 10.1002/jcp.25953   open full text
• Leptin accelerates the pathogenesis of heterotopic ossification in rat tendon tissues via mTORC1 signaling.
  Huaji Jiang, Yuhui Chen, Guorong Chen, Xinggui Tian, Jiajun Tang, Lei Luo, Minjun Huang, Bin Yan, Xiang Ao, Wen Zhou, Liping Wang, Xiaochun Bai, Zhongmin Zhang, Liang Wang, Cory J. Xian.
  Journal of Cellular Physiology. April 13, 2017
  Leptin, an adipocyte‐derived cytokine associated with bone metabolism, is believed to play a critical role in the pathogenesis of heterotopic ossification (HO). The effect and underlying action mechanism of leptin were investigated on osteogenic differentiation of tendon‐derived stem cells (TDSCs) in vitro and the HO formation in rat tendons. Isolated rat TDSCs were treated with various concentrations of leptin in the presence or absence of mTORC1 signaling specific inhibitor rapamycin in vitro. A rat model with Achilles tenotomy was employed to evaluate the effect of leptin on HO formation together with or without rapamycin treatment. In vitro studies with TDSCs showed that leptin increased the expression of osteogenic biomarkers (alkaline phosphatase, runt‐related transcription factor 2, osterix, osteocalcin) and enhanced mineralization of TDSCs via activating the mTORC1 signal pathway (as indicated by phosphorylation of p70 ribosomal S6 kinase 1 and p70 ribosomal S6). However, mTORC1 signaling blockade with rapamycin treatment suppressed leptin‐induced osteogenic differentiation and mineralization. In vivo studies showed that leptin promoted HO formation in the Achilles tendon after tenotomy, and rapamycin treatment blocked leptin‐induced HO formation. In conclusion, leptin can promote TDSC osteogenic differentiation and heterotopic bone formation via mTORC1 signaling in both in vitro and in vivo models, which provides a new potential therapeutic target for HO prevention. This article is protected by copyright. All rights reserved

  April 13, 2017   doi: 10.1002/jcp.25955   open full text
• Complex Osteoclastogenic Inductive Effects of Nicotine Over Hydroxyapatite.
  J. Costa‐Rodrigues, I. Rocha, M.H. Fernandes.
  Journal of Cellular Physiology. April 13, 2017
  Cigarette smoke is associated to pathological weakening of bone tissue, being considered an important playmaker in conditions such as osteoporosis and periodontal bone loss. In addition, it is also associated with an increased risk of failure in bone regeneration strategies. The present work aimed to characterize the effects of nicotine on human osteoclastogenesis over a hydroxyapatite substrate. Osteoclast precursors were maintained in the absence or presence of the osteoclastogenesis enhancers M‐CSF and RANKL, and were further treated with nicotine levels representative of the concentrations observed in the plasma and saliva of smokers. It was observed that nicotine at low concentrations elicit an increase in osteoclast differentiation, but only in the presence of M‐CSF and RANKL it was also able to significantly increase the resorbing ability of osteoclasts. A slight downregulation of NFkB pathway and an increase in the production of TNF‐α and, particularly PGE2, were involved in the observed effects of nicotine. At high concentrations, nicotine revealed cytotoxic effects, causing a decrease in cell density. In conclusion, nicotine at levels found in the plasma of the smokers, has the ability to act directly on osteoclast precursors, inducing its osteoclastogenic differentiation. The stimulatory behavior appears to be dependent on the stage of osteoclastic differentiation of the precursor cells, which means, in the absence of M‐CSF and RANKL, it only favors the initial stages of osteoclast differentiation, while in the presence of the growth factors, a significant increase in their resorbing ability is also achieved. This article is protected by copyright. All rights reserved

  April 13, 2017   doi: 10.1002/jcp.25956   open full text
• Pin1, the Master Orchestrator of Bone Cell Differentiation.
  Rabia Islam, Won‐Joon Yoon, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. April 12, 2017
  Pin1 is an enzyme that specifically recognizes the peptide bond between phosphorylated serine or threonine (pS/pT‐P) and proline. This recognition causes a conformational change of its substrate, which further regulates downstream signaling. Pin1−/− mice show developmental bone defects and reduced mineralization. Pin1 targets RUNX2 (Runt‐Related Transcription Factor 2), SMAD1/5, and β‐catenin in the FGF, BMP, and WNT pathways, respectively. Pin1 has multiple roles in the crosstalk between different anabolic bone signaling pathways. For example, it controls different aspects of osteoblastogenesis and increases the transcriptional activity of Runx2, both directly and indirectly. Pin1 also influences osteoclastogenesis at different stages by targeting PU.1 (Purine‐rich nucleic acid binding protein 1), C‐FOS, and DC‐STAMP. The phenotype of Pin1−/− mice has led to the recent identification of multiple roles of Pin1 in different molecular pathways in bone cells. These roles suggest that Pin1 can be utilized as an efficient drug target in congenital and acquired bone diseases. J. Cell. Physiol. 232: 2339–2347, 2017. © 2016 Wiley Periodicals, Inc. Pin1 is a peptidyl prolyl cis–trans isomerase that isomerizes only phosphorylated Ser/Thr‐Pro motifs. Upon binding, Pin1 mediates structural modifications resulting in dramatic changes in substrate activity. In this review, we have summarized the known roles of Pin1 in molecular regulation of bone cell differentiation. Pin1 binds to Runx2, Smad1/5, and β‐catenin, which act downstream of the FGF, BMP, and Wnt signaling pathways, respectively. Binding of Pin1 increases the stability and transcriptional activity of Runx2 and Smad1/5. Pin1 also inhibits the nuclear export of β‐catenin and increases the level of transcriptionally active nuclear β‐catenin.

  April 12, 2017   doi: 10.1002/jcp.25442   open full text
• Human White Adipocytes Convert Into “Rainbow” Adipocytes In Vitro.
  Giulia Maurizi, Antonella Poloni, Domenico Mattiucci, Spartaco Santi, Angela Maurizi, Valerio Izzi, Angelica Giuliani, Stefania Mancini, Maria Cristina Zingaretti, Jessica Perugini, Ilenia Severi, Massimo Falconi, Marco Vivarelli, Maria Rita Rippo, Silvia Corvera, Antonio Giordano, Pietro Leoni, Saverio Cinti.
  Journal of Cellular Physiology. April 12, 2017
  White adipocytes are plastic cells able to reversibly transdifferentiate into brown adipocytes and into epithelial glandular cells under physiologic stimuli in vivo. These plastic properties could be used in future for regenerative medicine, but are incompletely explored in their details. Here, we focused on plastic properties of human mature adipocytes (MA) combining gene expression profile through microarray analysis with morphologic data obtained by electron and time lapse microscopy. Primary MA showed the classic morphology and gene expression profile of functional mature adipocytes. Notably, despite their committed status, MA expressed high levels of reprogramming genes. MA from ceiling cultures underwent transdifferentiation toward fibroblast‐like cells with a well‐differentiated morphology and maintaining stem cell gene signatures. The main morphologic aspect of the transdifferentiation process was the secretion of large lipid droplets and the development of organelles necessary for exocrine secretion further supported the liposecretion process. Of note, electron microscope findings suggesting liposecretion phenomena were found also in explants of human fat and rarely in vivo in fat biopsies from obese patients. In conclusion, both MA and post‐liposecretion adipocytes show a well‐differentiated phenotype with stem cell properties in line with the extraordinary plasticity of adipocytes in vivo. J. Cell. Physiol. 232: 2887–2899, 2017. © 2016 Wiley Periodicals, Inc. Plastic properties of human mature adipocytes (MA) have been explored combining gene expression profile made by microarray analysis with morphologic data obtained by electron and time lapse microscopy. Mature adipocytes from ceiling cultures underwent transdifferentiation toward fibroblast‐like cells with a well‐differentiated morphology and maintaining stem cell gene signatures.

  April 12, 2017   doi: 10.1002/jcp.25743   open full text
• Serotonin transporter protects the placental cells against apoptosis in caspase 3‐independent pathway.
  Coedy Hadden, Tariq Fahmi, Anthonya Cooper, Alena V. Savenka, Vladimir V. Lupashin, Drucilla J. Roberts, Luc Maroteaux, Sylvie Hauguel‐de Mouzon, Fusun Kilic.
  Journal of Cellular Physiology. April 12, 2017
  Serotonin (5‐HT) and its specific transporter, SERT play important roles in pregnancy. Using placentas dissected from 18d gestational SERT‐knock out (KO), peripheral 5‐HT (TPH1)‐KO, and wild‐type (WT) mice, we explored the role of 5‐HT and SERT in placental functions in detail. An abnormal thick band of fibrosis and necrosis under the giant cell layer in SERT‐KO placentas appeared only moderately in TPH1‐KO and minimally present in WT placentas. The majority of the changes were located at the junctional zone of the placentas in SERT. The etiology of these findings was tested with TUNEL assays. The placentas from SERT‐KO and TPH1‐KO showed 49‐ and 8‐fold increase in TUNEL‐positive cells without a concurrent change in the DNA repair or cell proliferation compared to WT placentas. While the proliferation rate in the embryos of TPH1‐KO mice was 16‐fold lower than the rate in gestational age matched embryos of WT or SERT‐KO mice. These findings highlight an important role of continuous 5‐HT signaling on trophoblast cell viability. SERT may contribute to protecting trophoblast cells against cell death via terminating the 5‐HT signaling which changes cell death ratio in trophoblast as well as proliferation rate in embryos. However, the cell death in SERT‐KO placentas is in caspase 3‐independent pathway.

  April 12, 2017   doi: 10.1002/jcp.25812   open full text
• Dendritic Cell‐Specific Transmembrane Protein (DC‐STAMP) Regulates Osteoclast Differentiation via the Ca2+/NFATc1 Axis.
  Ya‐Hui Chiu, Edward Schwarz, Dongge Li, Yuexin Xu, Tzong‐Ren Sheu, Jinbo Li, Karen L. de Mesy Bentley, Changyong Feng, Baoli Wang, Jhih‐Cheng Wang, Liz Albertorio‐Saez, Ronald Wood, Minsoo Kim, Wensheng Wang, Christopher T. Ritchlin.
  Journal of Cellular Physiology. April 12, 2017
  DC‐STAMP is a multi‐pass transmembrane protein essential for cell–cell fusion between osteoclast precursors during osteoclast (OC) development. DC‐STAMP−/− mice have mild osteopetrosis and form mononuclear cells with limited resorption capacity. The identification of an Immunoreceptor Tyrosine‐based Inhibitory Motif (ITIM) on the cytoplasmic tail of DC‐STAMP suggested a potential signaling function. The absence of a known DC‐STAMP ligand, however, has hindered the elucidation of downstream signaling pathways. To address this problem, we engineered a light‐activatable DC‐STAMP chimeric molecule in which light exposure mimics ligand engagement that can be traced by downstream Ca2+ signaling. Deletion of the cytoplasmic ITIM resulted in a significant elevation in the amplitude and duration of intracellular Ca2+ flux. Decreased NFATc1 expression in DC‐STAMP−/− cells was restored by DC‐STAMP over‐expression. Multiple biological phenotypes including cell–cell fusion, bone erosion, cell mobility, DC‐STAMP cell surface distribution, and NFATc1 nuclear translocation were altered by deletion of the ITIM and adjacent amino acids. In contrast, mutations on each of the tyrosine residues surrounding the ITIM showed no effect on DC‐STAMP function. Collectively, our results suggest that the ITIM on DC‐STAMP is a functional motif that regulates osteoclast differentiation through the NFATc1/Ca2+ axis. J. Cell. Physiol. 232: 2538–2549, 2017. © 2016 Wiley Periodicals, Inc.

  April 12, 2017   doi: 10.1002/jcp.25638   open full text
• Contributions of 3D Cell Cultures for Cancer Research.
  Maddaly Ravi, Aarthi Ramesh, Aishwarya Pattabhi.
  Journal of Cellular Physiology. April 12, 2017
  Cancer cell lines have contributed immensely in understanding the complex physiology of cancers. They are excellent material for studies as they offer homogenous samples without individual variations and can be utilised with ease and flexibility. Also, the number of assays and end‐points one can study is almost limitless; with the advantage of improvising, modifying or altering several variables and methods. Literally, a new dimension to cancer research has been achieved by the advent of 3Dimensional (3D) cell culture techniques. This approach increased many folds the ways in which cancer cell lines can be utilised for understanding complex cancer biology. 3D cell culture techniques are now the preferred way of using cancer cell lines to bridge the gap between the ‘absolute in vitro’ and ‘true in vivo’. The aspects of cancer biology that 3D cell culture systems have contributed include morphology, microenvironment, gene and protein expression, invasion/migration/metastasis, angiogenesis, tumour metabolism and drug discovery, testing chemotherapeutic agents, adaptive responses and cancer stem cells. We present here, a comprehensive review on the applications of 3D cell culture systems for these aspects of cancers. J. Cell. Physiol. 232: 2679–2697, 2017. © 2016 Wiley Periodicals, Inc. 3D cell cultures are gaining ground for several applications, of which cancer research is an important area. Several aspects of cancer physiology have been unraveled using these cultures. Also, 3D cell cultures provide realistic material for drug discovery and biomarker discovery.

  April 12, 2017   doi: 10.1002/jcp.25664   open full text
• The Role of Exercise and TFAM in Preventing Skeletal Muscle Atrophy.
  Nicholas T. Theilen, George H. Kunkel, Suresh C. Tyagi.
  Journal of Cellular Physiology. April 12, 2017
  Skeletal muscle atrophy is the consequence of protein degradation exceeding protein synthesis. This arises for a multitude of reasons including the unloading of muscle during microgravity, post‐surgery bedrest, immobilization of a limb after injury, and overall disuse of the musculature. The development of therapies prior to skeletal muscle atrophy settings to diminish protein degradation is scarce. Mitochondrial dysfunction is associated with skeletal muscle atrophy and contributes to the induction of protein degradation and cell apoptosis through increased levels of ROS observed with the loss of organelle function. ROS binds mtDNA, leading to its degradation and decreasing functionality. Mitochondrial transcription factor A (TFAM) will bind and coat mtDNA, protecting it from ROS and degradation while increasing mitochondrial function. Exercise stimulates cell signaling pathways that converge on and increase PGC‐1α, a well‐known activator of the transcription of TFAM and mitochondrial biogenesis. Therefore, in the present review we are proposing, separately, exercise and TFAM treatments prior to atrophic settings (muscle unloading or disuse) alleviate skeletal muscle atrophy through enhanced mitochondrial adaptations and function. Additionally, we hypothesize the combination of exercise and TFAM leads to a synergistic effect in targeting mitochondrial function to prevent skeletal muscle atrophy. J. Cell. Physiol. 232: 2348–2358, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by © 2016 Wiley Periodicals, Inc. Skeletal muscle atrophy via unloading and disuse of the musculature is the consequence of protein degradation exceeding protein synthesis. This state is correlated with increased markers of mitochondrial dysfunction. We hypothesize targeting increased mitochondrial function through exercise and mitochondrial transcription factor A treatments will diminish skeletal muscle atrophy after a period of muscle unloading and disuse.

  April 12, 2017   doi: 10.1002/jcp.25737   open full text
• Homocysteine as a Pathological Biomarker for Bone Disease.
  Jyotirmaya Behera, Jyoti Bala, Mohammed Nuru, Suresh C. Tyagi, Neetu Tyagi.
  Journal of Cellular Physiology. April 12, 2017
  In the last few decades, perturbation in methyl‐group and homocysteine (Hcy) balance have emerged as independent risk factors in a number of pathological conditions including neurodegenerative disease, cardiovascular dysfunction, cancer development, autoimmune disease, and kidney disease. Recent studies report Hcy to be a newly recognized risk factor for osteoporosis. Elevated Hcy levels are known to modulate osteoclastgenesis by causing detrimental effects on bone via oxidative stress induced metalloproteinase‐mediated extracellular matrix degradation and decrease in bone blood flow. Evidence from previous studies also suggests that the decreased chondrocytes mediated bone mineralization in chick limb‐bud mesenchymal cells and during the gestational period of ossification in rat model. However, Hcy imbalance and its role in bone loss, regression in vascular invasion, and osteoporosis, are not clearly understood. More investigations are required to explore the complex interplay between Hcy imbalance and onset of bone disease progression. This article reviews the current body of knowledge on regulation of Hcy mediated oxidative stress and its role in bone remodeling, vascular blood flow and progression of bone disease. J. Cell. Physiol. 232: 2704–2709, 2017. © 2016 Wiley Periodicals, Inc. This article reviews the current body of knowledge on regulation of Hcy‐mediated oxidative stress and its role in bone remodeling, vascular blood flow, and progression of bone disease.

  April 12, 2017   doi: 10.1002/jcp.25693   open full text
• Nuclear Localization of Diacylglycerol Kinase Alpha in K562 Cells Is Involved in Cell Cycle Progression.
  Alessandro Poli, Roberta Fiume, Gianluca Baldanzi, Daniela Capello, Stefano Ratti, Marco Gesi, Lucia Manzoli, Andrea Graziani, Pann‐Ghill Suh, Lucio Cocco, Matilde Y. Follo.
  Journal of Cellular Physiology. April 10, 2017
  Phosphatidylinositol (PI) signaling is an essential regulator of cell motility and proliferation. A portion of PI metabolism and signaling takes place in the nuclear compartment of eukaryotic cells, where an array of kinases and phosphatases localize and modulate PI. Among these, Diacylglycerol Kinases (DGKs) are a class of phosphotransferases that phosphorylate diacylglycerol and induce the synthesis of phosphatidic acid. Nuclear DGKalpha modulates cell cycle progression, and its activity or expression can lead to changes in the phosphorylated status of the Retinoblastoma protein, thus, impairing G1/S transition and, subsequently, inducing cell cycle arrest, which is often uncoupled with apoptosis or autophagy induction. Here we report for the first time not only that the DGKalpha isoform is highly expressed in the nuclei of human erythroleukemia cell line K562, but also that its nuclear activity drives K562 cells through the G1/S transition during cell cycle progression. J. Cell. Physiol. 232: 2550–2557, 2017. © 2016 Wiley Periodicals, Inc.

  April 10, 2017   doi: 10.1002/jcp.25642   open full text
• Arabinogalactan Proteins From Baobab and Acacia Seeds Influence Innate Immunity of Human Keratinocytes In Vitro.
  Abderrakib Zahid, Julie Despres, Magalie Benard, Eric Nguema‐Ona, Jerome Leprince, David Vaudry, Christophe Rihouey, Maité Vicré‐Gibouin, Azeddine Driouich, Marie‐Laure Follet‐Gueye.
  Journal of Cellular Physiology. April 10, 2017
  Plant derived arabinogalactan proteins (AGP) were repeatedly confirmed as immunologically as well as dermatologically active compounds. However, little is currently known regarding their potential activity toward skin innate immunity. Here, we extracted and purified AGP from acacia (Acacia senegal) and baobab (Adansonia digitata) seeds to investigate their biological effects on the HaCaT keratinocyte cell line in an in vitro system. While AGP from both sources did not exhibit any cytotoxic effect, AGP from acacia seeds enhanced cell viability. Moreover, real‐time quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) analysis showed that AGP extracted from both species induced a substantial overexpression of hBD‐2, TLR‐5, and IL1‐α genes. These data suggest that plant AGP, already known to control plant defensive processes, could also modulate skin innate immune responses. J. Cell. Physiol. 232: 2558–2568, 2017. © 2016 Wiley Periodicals, Inc. Real‐time quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) analysis showed that AGP extracted from acacia or baobab seeds induced a substantial overexpression of hBD‐2, TLR‐5, and IL1‐α genes. These data suggest that plant AGP, already known to control plant defensive processes, could also modulate skin innate immune responses.

  April 10, 2017   doi: 10.1002/jcp.25646   open full text
• Anti‐Tumor Effects of Cardiac Glycosides on Human Lung Cancer Cells and Lung Tumorspheres.
  Vivek Kaushik, Juan Sebastian Yakisich, Neelam Azad, Yogesh Kulkarni, Rajkumar Venkatadri, Clayton Wright, Yon Rojanasakul, Anand Krishnan V. Iyer.
  Journal of Cellular Physiology. April 10, 2017
  Lung cancer is a leading cause of cancer‐related death in the United States. Although several drugs have been developed that target individual biomarkers, their success has been limited due to intrinsic or acquired resistance for the specific targets of such drugs. A more effective approach is to target multiple pathways that dictate cancer progression. Cardiac glycosides demonstrate such multimodal effects on cancer cell survival, and our aim was to evaluate the effect of two naturally occurring monosaccaridic cardiac glycosides—Convallatoxin and Peruvoside on lung cancer cells. Although both drugs had significant anti‐proliferative effects on H460 and Calu‐3 lung cancer cells, Convallatoxin demonstrated twofold higher activity as compared to Peruvoside using both viability and colony forming assays, suggesting a role for the aglycone region in dictating drug potency. The tumor suppressor p53 was found to be important for action of both drugs—p53‐underexpressing cells were less sensitive as compared to p53‐positive H460 cells. Further, assessment of p53‐underexpressing H460 cells showed that drugs were able to arrest cells in the G0/G1 phase of the cell cycle in a dose‐dependent manner. Both drugs significantly inhibited migration and invasion of cancer cells and decreased the viability of floating tumorspheres. An assessment of intracellular pathways indicated that both drugs were able to modulate proteins that are involved in apoptosis, autophagy, cell cycle, proliferation, and EMT. Our data suggest, a promising role for cardiac glycosides in lung cancer treatment, and provides impetus for further investigation of the anti‐cancer potential of this class of drugs. J. Cell. Physiol. 232: 2497–2507, 2017. © 2016 Wiley Periodicals, Inc. The cardiac glycosides convallatoxin and peruvoside exert potent anti‐cancer effects in lung cancer cells. A significant inhibition of cell migration, invasion, and lung tumorsphere formation was observed in response to both drugs.

  April 10, 2017   doi: 10.1002/jcp.25611   open full text
• The Role of Calcium‐Sensing Receptors in Endothelin‐1‐Dependent Effects on Adult Rat Ventricular Cardiomyocytes: Possible Contribution to Adaptive Myocardial Hypertrophy.
  Elena Dyukova, Rolf Schreckenberg, Christoph Arens, Guzel Sitdikova, Klaus‐Dieter Schlüter.
  Journal of Cellular Physiology. April 10, 2017
  Nitric oxide (NO)‐deficiency as it occurs during endothelial dysfunction activates the endothelin‐1 (ET‐1) system and increases the expression of receptor activity modifying protein (RAMP)‐1 that acts as a chaperon for calcium‐sensing receptors (CaR) that have recently been identified to improve cardiac function. Here, we hypothesized that ET‐1 increases the cardiac expression of CaR and thereby induces an adaptive type of hypertrophy. Expressions of RAMP‐1, endothelin receptors, and CaR were analyzed by RT‐PCR in left ventricular tissues of L‐NAME‐treated rats. Effects of ET‐1 on CaR expression and cell function (load free cell shortening) were analyzed in adult rat ventricular cardiomyocytes. siRNA directed against CaR and RAMP‐1 was used to investigate a causal relationship. PD142893 and BQ788 were used to dissect the contribution of ETB1, ETB2, and ETA receptors. Non‐specific NO synthase inhibition with L‐Nitro arginine methyl ester (L‐NAME) caused a cardiac upregulation of ETB receptors and CaR suggesting a paracrine effect of ET‐1 on cardiomyocytes. Indeed, ET‐1 induced the expression of CaR in cultured cardiomyocytes. Under these conditions, cardiomyocytes increased cell size (hypertrophy) but maintained normal function. Inhibition of ETA and ETB1 receptors led to ET‐1‐dependent reduction in cell shortening and attenuated up‐regulation of CaR. Down‐regulation of RAMP‐1 reduced CaR responsiveness. In conclusion, ET‐1 causes an adaptive type of hypertrophy by up‐regulation of CaR in cardiomyocytes via ETA and/or ETB1 receptors. J. Cell. Physiol. 232: 2508–2518, 2017. © 2016 Wiley Periodicals, Inc. Current manuscript is the first report indicating the ET‐1‐dependent upregulation of CaR receptor in cardiomyocytes and it shows that this upregulation allows cardiac cells to maintain proper function. Second new finding indicates involvement of ETA and /or ETB1 receptors in adaptive hypertrophy in cardiomyocytes, whereas ETB2 receptors activation leads to mal‐adaptive effect. Finally, chaperone activity of RAMP1 is required for the functional coupling of CaR in cardiomyocytes.

  April 10, 2017   doi: 10.1002/jcp.25612   open full text
• Lkb1 regulation of skeletal muscle development, metabolism and muscle progenitor cell homeostasis.
  Tizhong Shan, Ziye Xu, Jiaqi Liu, Weiche Wu, Yizhen Wang.
  Journal of Cellular Physiology. April 10, 2017
  Liver kinase B1 (Lkb1), also named as Serine/Threonine protein kinase 11 (STK11), is a serine/threonine kinase that plays crucial roles in various cellular processes including cell survival, cell division, cellular polarity, cell growth, cell differentiation, and cell metabolism. In metabolic tissues, Lkb1 regulates glucose homeostasis and energy metabolism through phosphorylating and activating the AMPK subfamily proteins. In skeletal muscle, Lkb1 affects muscle development and postnatal growth, lipid and fatty acid oxidation, glucose metabolism, and insulin sensitivity. Recently, the regulatory roles of Lkb1 in regulating division, self‐renew, proliferation, and differentiation of skeletal muscle progenitor cells have been reported. In this review, we discuss the roles of Lkb1 in regulating skeletal muscle progenitor cell homeostasis and skeletal muscle development and metabolism. In skeletal muscle, Lkb1 affects muscle development and postnatal growth, lipid, and fatty acid oxidation, glucose metabolism and insulin sensitivity. In skeletal muscle progenitor cells, Lkb1 regulates cell division, self‐renew, proliferation, and differentiation. Here, we review the role of Lkb1 in skeletal muscle development, metabolism and muscle progenitor cell homeostasis.

  April 10, 2017   doi: 10.1002/jcp.25786   open full text
• Stimulatory actions of a novel thiourea derivative on large‐conductance, calcium‐activated potassium channels.
  Sheng‐Nan Wu, Jyh‐Haur Chern, Santai Shen, Hwei‐Hisen Chen, Ying‐Ting Hsu, Chih‐Chin Lee, Ming‐Huan Chan, Ming‐Chi Lai, Feng‐Shiun Shie.
  Journal of Cellular Physiology. April 10, 2017
  In this study, we examine whether an anti‐inflammatory thiourea derivative, compound #326, actions on ion channels. The effects of compound #326 on Ca2+‐activated K+ channels were evaluated by patch‐clamp recordings obtained in cell‐attached, inside‐out or whole‐cell configuration. In pituitary GH3 cells, compound #326 increased the amplitude of Ca2+‐activated K+ currents (IK(Ca)) with an EC50 value of 11.6 μM, which was reversed by verruculogen, but not tolbutamide or TRAM‐34. Under inside‐out configuration, a bath application of compound #326 raised the probability of large‐conductance Ca2+‐activated K+ (BKCa) channels. The activation curve of BKCa channels was shifted to less depolarised potential with no modification of the gating charge of the curve; consequently, the difference of free energy was reduced in the presence of this compound. Compound #326‐stimulated activity of BKCa channels is explained by a shortening of mean closed time, despite its inability to alter single‐channel conductance. Neither delayed‐rectifier nor erg‐mediated K+ currents was modified. Compound #326 decreased the peak amplitude of voltage‐gated Na+ current with no clear change in the overall current–voltage relationship of this current. In HEK293T cells expressing α‐hSlo, compound #326 enhanced BKCa channels effectively. Intriguingly, the inhibitory actions of compound #326 on interleukin 1β in lipopolysaccharide‐activated microglia were significantly reversed by verruculogen, whereas BKCa channel inhibitors suppressed the expressions of inducible nitric oxide synthase. The BKCa channels could be an important target for compound #326 if similar in vivo results occur, and the multi‐functionality of BKCa channels in modulating microglial immunity merit further investigation. Compound #326‐stimulated activity of BKCa channels is explained by a shortening of mean closed time, despite its inability to alter single‐channel conductance. Intriguingly, the inhibitory actions of compound #326 on interleukin 1β in lipopolysaccharide‐activated microglia were significantly reversed by verruculogen, whereas BKCa channel inhibitors suppressed the expressions of inducible nitric oxide synthase.

  April 10, 2017   doi: 10.1002/jcp.25788   open full text
• Inhibitory effect of blue light emitting diode on migration and invasion of cancer cells.
  Phil‐Sun Oh, Hyun‐Soo Kim, Eun‐Mi Kim, Hyosook Hwang, Hyang Hwa Ryu, SeokTae Lim, Myung‐Hee Sohn, Hwan‐Jeong Jeong.
  Journal of Cellular Physiology. April 10, 2017
  The aim of this study was to determine the effects and molecular mechanism of blue light emitting diode (LED) in tumor cells. A migration and invasion assay for the metastatic behavior of mouse colon cancer CT‐26 and human fibrosarcoma HT‐1080 cells was performed. Cancer cell migration‐related proteins were identified by obtaining a 2‐dimensional gel electrophoresis (2‐DE) in total cellular protein profile of blue LED‐irradiated cancer cells, followed by matrix‐assisted laser desorption/ionization‐time of flight (MALDI‐TOF) analysis of proteins. Protein levels were examined by immunoblotting. Irradiation with blue LED inhibited CT‐26 and HT‐1080 cell migration and invasion. The anti‐metastatic effects of blue LED irradiation were associated with inhibition of matrix metalloproteinase (MMP)‐2 and MMP‐9 expression. P38 MAPK phosphorylation was increased in blue LED‐irradiated CT‐26 and HT‐1080 cells, but was inhibited after pretreatment with SB203580, a specific inhibitor of p38 MAPK. Inhibition of p38 MAPK phosphorylation by SB203580 treatment increased number of migratory cancer cells in CT‐26 and HT‐1080 cells, indicating that blue LED irradiation inhibited cancer cell migration via phosphorylation of p38 MAPK. Additionally blue LED irradiation of mice injected with CT‐26 cells expressing luciferase decreased early stage lung metastasis compared to untreated control mice. These results indicate that blue LED irradiation inhibits cancer cell migration and invasion in vitro and in vivo.

  April 10, 2017   doi: 10.1002/jcp.25805   open full text
• Let‐7i‐Induced Atg4B Suppression Is Essential for Autophagy of Placental Trophoblast in Preeclampsia.
  Yinyan Xu, Xinyan Huang, Juan Xie, Yanni Chen, Jing Fu, Li Wang.
  Journal of Cellular Physiology. April 10, 2017
  Autophagy, identified as type II programmed cell death, has already been known to be involved in the pathophysiology of preeclampsia (PE), which is a gestational disease with high morbidity. The present study aims to investigate the functional role of let‐7i, a miRNA, in trophoblastic autophagy. Placental tissue used in this study was collected from patients with severe preeclampsia (SPE) or normal pregnant women. A decreased level of let‐7i was found in placenta of SPE. In addition, autophagic vacuoles were observed in SPE and the expression of microtubule associated protein 1 light chain 3 (LC3) II/I was elevated. In vitro, let‐7i mimics suppressed the autophagic activities in human HTR‐8/SVneo trophoblast cell line (HTR‐8) and human placental choriocarcinoma cell line JEG‐3, whereas let‐7i inhibitor enhanced the activities. As a potential target of let‐7i, autophagy‐related 4B cysteine peptidase (Atg4B) had an increased expression level in SPE. As expected, the increased expression of Atg4B was negatively regulated by let‐7i using dual luciferase reporter assay. Furthermore, these trophoblast‐like cells transfected with the let‐7i mimic or inhibitors resulted in a significant change of Atg4B in both mRNA and protein level. More importantly, Atg4B overexpression could partly reverse let‐7i mimic‐reduced LC3II/I levels; whereas Atg4B silencing partly attenuated let‐7i inhibitor‐induced the level of LC3II/I expression. Taken together, these findings suggest that let‐7i is able to regulate autophagic activity via regulating Atg4B expression, which might contribute to the pathogenesis of PE. J. Cell. Physiol. 232: 2581–2589, 2017. © 2016 Wiley Periodicals, Inc. This study provided insight into the role of let‐7i in the pathophysiology of PE. Our study demonstrates for the first time that a novel let‐7i‐Atg4B‐autophagy signaling pathway plays a key role in PE. This finding suggests that miRNAs are key molecules involved in autophagic activity, and might be regard as a new therapeutic target against PE.

  April 10, 2017   doi: 10.1002/jcp.25661   open full text
• KCNQ1 variants associate with hypertension in type 2 diabetes and affect smooth muscle contractility in vitro.
  Kuo‐Chin Huang, Te‐Mao Li, Xiang Liu, Jin‐Hua Chen, Wen‐Kuei Chien, Yi‐Tzone Shiao, Hsinyi Tsang, Ting‐Hsu Lin, Chiu‐Chu Liao, Shao‐Mei Huang, Ju‐Pi Li, Cheng‐Wen Lin, Jung‐Chun Lin, Chih‐Chien Lin, Chih‐Ho Lai, Chi‐Fung Cheng, Wen‐Miin Liang, Chien‐Hui Hung, Ching‐Chu Chen, Ying‐Ju Lin, Fuu‐Jen Tsai.
  Journal of Cellular Physiology. April 10, 2017
  KCNQ1 encodes a potassium voltage‐gated channel and represents a susceptibility locus for type 2 diabetes mellitus (T2DM). Here, we explored the association between KCNQ1 polymorphisms and hypertension risk in individuals with T2DM, as well as the role of KCNQ1 in vascular smooth muscle cell contraction in vitro. To investigate the relationship between KCNQ1 and the risk of developing hypertension in patients with T2DM, we divided the T2DM cohort into hypertension (n = 452) and non‐hypertension (n = 541) groups. The Mann–Whitney U test, chi‐square test, and multivariate regression analyses were used to assess the clinical characteristics and genotypic frequencies. In vitro studies utilized the rat aortic smooth muscle A10 cell line. Patients in the hypertension group were significantly older at the time of enrollment and had higher levels of body mass index, waist‐to‐hip ratio, and triglyceride than those in the non‐hypertension group. The KCNQ1 rs3864884 and rs12576239 genetic variants were associated with hypertension in T2DM. KCNQ1 expression was lower in the individuals with the CC versus the CT and TT genotypes. Smooth muscle cell contractility was inhibited by treatment with a KCNQ1 inhibitor. These results suggest that KCNQ1 might be associated with hypertension in individuals with T2DM. The KCNQ1 rs3864884 and rs12576239 genetic variants were associated with hypertension in T2DM. Smooth muscle cell contractility was inhibited by treatment with a KCNQ1 inhibitor. KCNQ1 might be associated with hypertension in individuals with T2DM.

  April 10, 2017   doi: 10.1002/jcp.25775   open full text
• Irx3 and Bmp2 regulate mouse mesenchymal cell chondrogenic differentiation in both a Sox9‐dependent and ‐independent manner.
  Yoshihiro Tamamura, Kenichi Katsube, Hisashi Mera, Maki Itokazu, Shigeyuki Wakitani.
  Journal of Cellular Physiology. April 10, 2017
  Sox9, a master regulator of cartilage development, controls the cell fate decision to differentiate from mesenchymal to chondrogenic cells. In addition, Sox9 regulates the proliferation and differentiation of chondrocytes, as well as the production of cartilage‐specific proteoglycans. The existence of Sox9‐independent mechanisms in cartilage development remains to be determined. Here, we attempted to identify genes involved in such putative mechanisms via microarray analysis using a mouse chondrogenic cell line, N1511. We first focused on transcription factors that exhibited upregulated expression following Bmp2 treatment, which was not altered by subsequent treatment with Sox9 siRNA. Among these, we selected positive regulators for chondrogenesis and identified Iroquois‐related homeobox 3 (Irx3) as one of the candidate genes. Irx3 expression gradually increased with chondrocyte terminal differentiation in a reciprocal manner to Sox9 expression, and promoted the chondrogenic differentiation of mesenchymal cells upon Bmp2 treatment. Furthermore, Irx3 partially rescued impaired chondrogenesis by upregulating the expression of epiphycan and lumican under reduced Sox9 expression. Finally, Irx3 was shown to act in concert with Bmp2 signaling to activate the p38 MAPK pathway, which in turn stimulated Sox9 expression, as well as the expression of epiphycan and lumican in a Sox9‐independent manner. These results indicate that Irx3 represents a novel chondrogenic factor of mesenchymal cells, acts synergistically with Bmp2‐mediated signaling, and regulates chondrogenesis independent of the transcriptional machinery associated with Sox9‐mediated regulation. Irx3 cooperates with Bmp2 to regulate the expression of type II collagen and aggrecan in a Sox9‐dependent manner, whereas that of epiphycan and lumican in a Sox9‐independent manner in C3H10T1/2 cells.

  April 10, 2017   doi: 10.1002/jcp.25776   open full text
• Testosterone Rescues the De‐Differentiation of Smooth Muscle Cells Through Serum Response Factor/Myocardin.
  Carolina Leimgruber, Amado A. Quintar, Nahuel Peinetti, María V. Scalerandi, Juan P. Nicola, Joseph M. Miano, Cristina A. Maldonado.
  Journal of Cellular Physiology. April 10, 2017
  Prostatic smooth muscle cells (pSMCs) differentiation is a key factor for prostatic homeostasis, with androgens exerting multiple effects on these cells. Here, we demonstrated that the myodifferentiator complex Srf/Myocd is up‐regulated by testosterone in a dose‐dependent manner in primary cultures of rat pSMCs, which was associated to the increase in Acta2, Cnn1, and Lmod1 expressions. Blocking Srf or Myocd by siRNAs inhibited the myodifferentiator effect of testosterone. While LPS led to a dedifferentiated phenotype in pSMCs, characterized by down‐regulation of Srf/Myocd and smooth muscle cell (SMC)‐restricted genes, endotoxin treatment on Myocd‐overexpressing cells did not result in phenotypic alterations. Testosterone at a physiological dose was able to restore the muscular phenotype by normalizing Srf/Myocd expression in inflammation‐induced dedifferentiated pSMCs. Moreover, the androgen reestablished the proliferation rate and IL‐6 secretion increased by LPS. These results provide novel evidence regarding the myodifferentiating role of testosterone on SMCs by modulating Srf/Myocd. Thus, androgens preserve prostatic SMC phenotype, which is essential to maintain the normal structure and function of the prostate. J. Cell. Physiol. 232: 2806–2817, 2017. © 2016 Wiley Periodicals, Inc. Androgens favor a contractile phenotype on smooth muscle cells that relies on the Myocd/SRF myodifferentiator complex, with this role being critical for cellular homeostasis and pathophysiology.

  April 10, 2017   doi: 10.1002/jcp.25679   open full text
• TNF‐α promotes osteoclastogenesis through JNK signaling‐dependent induction of Semaphorin3D expression in estrogen‐deficiency induced osteoporosis.
  Chenglin Sang, Jiefeng Zhang, Yongxian Zhang, Fangjing Chen, Xuecheng Cao, Lei Guo.
  Journal of Cellular Physiology. April 10, 2017
  Tumor necrosis factor α (TNF‐α)‐induced osteoclast formation have been demonstrated to play an important role in the pathogenesis of estrogen deficiency‐mediated bone loss, but the exact mechanisms by which TNF‐α enhanced osteoclast differentiation were not fully elucidated. The class III semaphorins members were critical to regulate bone homeostasis. Here, we identified a novel mechanism whereby TNF‐α increasing Semaphorin3D expression contributes to estrogen deficiency‐induced osteoporosis. In this study, we found that Semaphorin3D expression was upregulated by TNF‐α during the process of RANKL‐induced osteoclast differentiation. Inhibition of Semaphorin3D in pre‐osteoclasts could attenuate the stimulatory effects of TNF‐α on osteoclast proliferation and differentiation. Mechanistically, blocking of the Jun N‐terminal kinase (JNK) signaling markedly rescued TNF‐α‐induced Semaphorin3D expression, suggesting that JNK signaling was involved in the regulation of Semaphorin3D expression by TNF‐α. In addition, silencing of Semaphorin3D in vivo could alleviate estrogen deficiency‐induced osteoporosis. Our results revealed a novel function for Semaphorin3D and suggested that increased Semaphorin3D may contribute to enhanced bone loss by increased TNF‐α in estrogen deficiency‐induced osteoporosis. Thus, Semaphorin3D may provide a potential therapeutic target for the treatment of estrogen‐deficiency induced osteoporosis. We found that TNF‐a could increase Sema3D expression in RANKL‐induced osteoclast formation. Upregulation of Sema3D expression was required for TNF‐a‐mediated osteoclast proliferation and differentiation. JNK signaling was involved in the regulation of Sema3D expression by TNF‐a.

  April 10, 2017   doi: 10.1002/jcp.25784   open full text
• Sensitivity of TP53‐Mutated Cancer Cells to the Phytoestrogen Genistein Is Associated With Direct Inhibition of Plk1 Activity.
  Sol‐Bi Shin, Sang‐Uk Woo, Young‐Won Chin, Young‐Joo Jang, Hyungshin Yim.
  Journal of Cellular Physiology. April 10, 2017
  Polo‐like kinase 1 (Plk1), a conserved Ser/Thr mitotic kinase, has been identified as a promising target for anticancer drug development because its overexpression is correlated with malignancy. Here, we found that genistein, an isoflavone, inhibits Plk1 kinase activity directly. Previously the mitotic disturbance phenomenon induced by treatment with genistein was not fully explained by its inhibitory effect on EGFR. In kinase profiling assays, it showed selectivity relative to a panel of kinases, including EGFR. Treatment with genistein induced cell death in a concentration‐dependent manner in cancer cells from diverse tissue origins, but not in non‐transformed cells such as hTERT‐RPE or MCF10A cells. We also observed that genistein tended to be more selective against cancer cells with mutations in the TP53 gene. TP53‐depeleted LNCaP and NCI‐H460 cells using shRNA targeting human TP53 were more sensitive to cell death by treatment of genistein. Furthermore, genistein induced mitotic arrest by inhibiting Plk1 activity and, consequently, led to mitotic catastrophe and apoptosis. These data suggest that genistein may be a promising anticancer drug candidate due to its inhibitory activity against Plk1 as well as EGFR and effectiveness toward cancer cells, especially those with p53‐mutation. J. Cell. Physiol. 232: 2818–2828, 2017. © 2016 Wiley Periodicals, Inc. The inhibitory activity of genistein on the Plk1 kinase was relatively selective to its activity against a panel of other kinases, including EGFR. Genistein had a cytotoxic effect in human cancer cells from diverse tissue origins, but not in non‐transformed cells. The sensitivity of cancer cells containing mutations in the TP53 gene to genistein tended to be higher when compared with that of cells with wild‐type TP53. Since around 50% of solid tumors have mutant TP53, the development of genistein as an anticancer drug would be valuable for treatment of such tumors.

  April 10, 2017   doi: 10.1002/jcp.25680   open full text
• A critical role for adiponectin‐mediated development of endometrial luminal epithelial cells during the peri‐implantation period of pregnancy.
  Whasun Lim, Myung Jin Choi, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. April 10, 2017
  Adiponectin is one of the adipokines in the collagen superfamily. It is secreted primarily by white adipocytes and influences reproductive processes including ovarian and uterine functions. Adiponectin regulates energy homeostasis, insulin sensitivity, and is anti‐inflammatory in various tissues. Its receptors (ADIPOR1 and ADIPOR2) are widely expressed in mammalian tissues, including porcine conceptuses and endometrial during the estrous cycle and peri‐implantation period of pregnancy. However, regulatory effects of adiponectin on endometrial epithelial cells are unknown. Therefore, we investigated the effects of parity on expression of ADIPOR1 and ADIPOR2 and the effects of adiponectin in the porcine endometrium during early pregnancy. Results of this study revealed robust expression of ADIPOR1 and ADIPOR2 in uterine luminal (LE) and glandular (GE) epithelia during early pregnancy and expression decreased as with increasing parity. For porcine luminal epithelial (pLE) cells, adiponectin enhanced proliferation, and increased phosphorylation of AKT, P70S6K, S6, ERK1/2, JNK, P38, and P90RSK in a time‐dependent manner. Moreover, the abundance of adiponectin‐activated signaling molecules were suppressed by pharmacological inhibitors including wortmannin, U0126, SP600125, and SB203580, respectively, in pLE cells. Furthermore, inhibition of each targeted signal transduction molecule influenced proliferation of adiponectin‐stimulated pLE cells. In addition, adiponectin inhibited tunicamycin‐induced endoplasmic reticulum (ER)‐stress through effects on ER stress regulated proteins in pLE cells. Collectively, these results suggest that adiponectin affects development of porcine uterine epithelia and reproductive performance through modulation of PI3K/AKT and MAPK cell signaling pathways. Adiponectin enhance uterine receptivity to implantation and placentation through PI3K/AKT and MAPK pathways for development of the porcine uterus and reproductive performance.

  April 10, 2017   doi: 10.1002/jcp.25768   open full text
• Anti‐Atherosclerotic Effects of Vitamins D and E in Suppression of Atherogenesis.
  Bahman Rashidi, Zahra Hoseini, Amirhossein Sahebkar, Hamed Mirzaei.
  Journal of Cellular Physiology. April 10, 2017
  Atherosclerosis is a progressive and multifactorial disease which occurs under the influence of various risk factors including endothelial dysfunction (ED), oxidative stress, and low‐density lipoprotein (LDL) oxidation. In contract to the initial hypotheses on the usefulness of vitamin E supplementation for cardiovascular disease prevention, large outcome trials showed consumption of vitamin E has no obvious effect on cardiovascular disease and, in some cases, it may even increase the rate of mortality. This seemingly unexpected finding may be due to the opposite effects of vitamin E compounds. Vitamin E is a group of compounds which have different and even opposing effects, yet in most of the studies, the exact consumed component of vitamin E is not determined. It appears that the combined consumption of gamma‐tocopherol, vitamin C, D, and tetrahydrobiopterin (BH4) may be extremely effective in both preventing atherogenesis and suppressing plaque development. In this regard, one of main issues is effect of vitamins E and D deficiency on microRNAs network in atherosclerosis. Various studies have indicated that miRNAs have key roles in atherosclerosis pathogenesis. The deficiency of vitamins E and D could provide a deregulation for miRNAs network and these events could lead to progression of atherosclerosis. Here, we highlighted a variety of mechanisms involve in the progression of atherosclerosis and effects of vitamins D and E on these mechanisms. Moreover, we summarized miRNAs involve in atherosclerosis and their regulation by vitamins E and D deficiency. J. Cell. Physiol. 9999: 1–9, 2016. © 2016 Wiley Periodicals, Inc.

  April 10, 2017   doi: 10.1002/jcp.25738   open full text
• Regulation of chondrocyte functions by transient receptor potential cation channel V6 in osteoarthritis.
  Tengfei Song, Jun Ma, Lei Guo, Peng Yang, Xuhui Zhou, Tianwen Ye.
  Journal of Cellular Physiology. April 10, 2017
  Transient receptor potential vanilloid (TRPV) channels function to maintain the dynamic balance of calcium signaling and calcium metabolism in bones. The goal of this study was to determine the potential role of TRPV6 in regulation of chondrocytes. The level of TRPV6 expression was analyzed by western blot in articular cartilage derived from the knee joints of osteoarthritis (OA) rat models and OA patients. Bone structure and osteoarthritic changes in the knee joints of TRPV6 knockout mice were examined using micro‐computed and histological analysis at the age of 6 and 12 months old. Furthermore, to investigate the effects of TRPV6 on chondrocyte extracellular matrix secretion, the release of matrix degrading enzymes, cell proliferation, and apoptosis, we decreased and increased TRPV6 expression in chondrocytes with lentiviral constructs encoding shRNA targeting TRPV6 and encoding TRPV6, respectively. The results showed that the level of TRPV6 expression in an OA rat model was markedly down‐regulated. TRPV6 knockout mice showed severe osteoarthritis changes, including cartilage fibrillation, eburnation, and loss of proteoglycans. In addition, deficiency of TRPV6 clearly affected chondrocyte function, such as extracellular matrix secretion, the release of matrix degrading enzymes, cell proliferation, and apoptosis. Taken together, our results implicated that TRPV6 channel, as a chondro‐protective factor, was involved in the pathogenesis of OA. The level of TRPV6 expression in an OA rat model was markedly down‐regulated. TRPV6 knockout mice showed severe osteoarthritic changes. Deficiency of TRPV6 clearly affected chondrocyte function.

  April 10, 2017   doi: 10.1002/jcp.25770   open full text
• Live Cell Imaging: Assessing the Phototoxicity of 488 and 546 nm Light and Methods to Alleviate it.
  Stephen Douthwright, Greenfield Sluder.
  Journal of Cellular Physiology. April 10, 2017
  In live cell imaging of fluorescent proteins, phototoxicity of the excitation light can be problematical. Cell death is obvious, but reduced cell viability can make the interpretation of observations error prone. We characterized the phototoxic consequences of 488 and 546 nm light on untransformed human cells and tested methods that have or could be used to alleviate photodamage. Unlabeled RPE1 cells were given single 0.5–2.5 min irradiations in early G1 from a mercury arc lamp on a fluorescence microscope. Four hundred eighty‐eight nanometer light produced a dose‐dependent decrease in the percentage of cells that progressed to mitosis, slowing of the cell cycle for some of those entering mitosis, and a ∼12% incidence of cell death for the highest dose. For 546 nm light we found a 10–15% reduction in the percentage of cells entering mitosis, no strong dose dependency, and a ∼2% incidence of cell death for the longest irradiations. For cells expressing GFP‐centrin1 or mCherry‐centrin1, fewer entered mitosis for each dose than unlabeled cells. For constant total dose 488 nm light irradiations of unlabeled cells, reducing the intensity 10‐fold or spreading the exposures out as a series of 10 sec pulses at 1 min intervals produced a minor and not consistent improvement in the percentage of cells entering mitosis. Reducing oxidative processes, by culturing at ∼3% oxygen or adding the reducing agent Trolox noticeably increased the fraction of cells entering mitosis. Thus, for long‐term imaging there can be value to using RFP constructs and for GFP‐tagged proteins reducing oxidative processes. J. Cell. Physiol. 232: 2461–2468, 2017. © 2016 Wiley Periodicals, Inc. We characterized the phototoxic consequences of 488 and 546 nm light on untransformed human cells. Four hundred eighty‐eight nanometer light produced a dose‐dependent decrease in the percentage of cells that progressed to mitosis, and slowing of the cell cycle for some of those entering mitosis and for 546 nm light, we found a 10–15% reduction in the percentage of cells entering mitosis, but no strong dose dependency. Reducing oxidative processes, by culturing at ∼3% oxygen or adding the reducing agent Trolox noticeably increased the fraction of cells entering mitosis.

  April 10, 2017   doi: 10.1002/jcp.25588   open full text
• Cardiotrophin‐1 Regulates Adipokine Production in 3T3‐L1 Adipocytes and Adipose Tissue From Obese Mice.
  Miguel López‐Yoldi, Beatriz Marcos‐Gomez, María Asunción Romero‐Lozano, Neira Sáinz, Jesús Prieto, Jose Alfredo Martínez, Matilde Bustos, Maria J. Moreno‐Aliaga.
  Journal of Cellular Physiology. April 10, 2017
  Cardiotrophin‐1 (CT‐1) belongs to the IL‐6 family of cytokines. Previous studies of our group revealed that CT‐1 is a key regulator of glucose and lipid metabolism. The aim of the present study was to analyze the in vitro and in vivo effects of CT‐1 on the production of several adipokines involved in body weight regulation, nutrient metabolism, and inflammation. For this purpose, 3T3‐L1 adipocytes were incubated with recombinant protein CT‐1 (rCT‐1) (1–40 ng/ml) for 1 and 18 h. Moreover, the acute effects of rCT‐1 administration (0.2 mg/kg, i.v.) for 30 min and 3 h on adipokines levels were also evaluated in high‐fat fed obese mice. In 3T3‐L1 adipocytes, rCT‐1 treatment downregulated the expression and secretion of leptin, resistin, and visfatin. However, rCT‐1 significantly stimulated apelin mRNA and secretion. rCT‐1 (18 h) also promoted the activation by phosphorylation of AKT, ERK 1/2, and STAT3. Interestingly, pre‐treatment with the PI3K inhibitor LY294002 reversed the stimulatory effects of rCT‐1 on apelin expression, suggesting that this pathway could be mediating the effects of rCT‐1 on apelin production. In contrast, acute administration of rCT‐1 (30 min and 3 h) to diet‐induced obese mice downregulated leptin and resistin, without significantly modifying apelin or visfatin mRNA in adipose tissue. Furthermore, CT‐1 null mice exhibited altered expression of adipokines in adipose tissue. The present study demonstrates that rCT‐1 modulates the production of adipokines in vitro and in vivo, suggesting that the regulation of the secretory function of adipocytes could be involved in the metabolic actions of this cytokine. J. Cell. Physiol. 232: 2469–2477, 2017. © 2016 Wiley Periodicals, Inc. The current study demonstrates that CT‐1 is capable of regulating the secretory pattern of adipokines by adipocytes, decreasing the production of pro‐inflammatory adipokines, such as leptin, resistin, and visfatin, while stimulating the secretion of apelin. These suggest that the beneficial actions of CT‐1 on glucose and lipid metabolism could be also related to its ability to control adipokine secretion and therefore the cross‐talk between adipose tissue and other key metabolic organs.

  April 10, 2017   doi: 10.1002/jcp.25590   open full text
• Overexpression of Large‐Conductance Calcium‐Activated Potassium Channels in Human Glioblastoma Stem‐Like Cells and Their Role in Cell Migration.
  Paolo Rosa, Luigi Sforna, Silvia Carlomagno, Giorgio Mangino, Massimo Miscusi, Mauro Pessia, Fabio Franciolini, Antonella Calogero, Luigi Catacuzzeno.
  Journal of Cellular Physiology. April 10, 2017
  Glioblastomas (GBMs) are brain tumors characterized by diffuse invasion of cancer cells into the healthy brain parenchyma, and establishment of secondary foci. GBM cells abundantly express large‐conductance, calcium‐activated potassium (BK) channels that are thought to promote cell invasion. Recent evidence suggests that the GBM high invasive potential mainly originates from a pool of stem‐like cells, but the expression and function of BK channels in this cell subpopulation have not been studied. We investigated the expression of BK channels in GBM stem‐like cells using electrophysiological and immunochemical techniques, and assessed their involvement in the migratory process of this important cell subpopulation. In U87‐MG cells, BK channel expression and function were markedly upregulated by growth conditions that enriched the culture in GBM stem‐like cells (U87‐NS). Cytofluorimetric analysis further confirmed the appearance of a cell subpopulation that co‐expressed high levels of BK channels and CD133, as well as other stem cell markers. A similar association was also found in cells derived from freshly resected GBM biopsies. Finally, transwell migration tests showed that U87‐NS cells migration was much more sensitive to BK channel block than U87‐MG cells. Our data show that BK channels are highly expressed in GBM stem‐like cells, and participate to their high migratory activity. J. Cell. Physiol. 232: 2478–2488, 2017. © 2016 Wiley Periodicals, Inc. We investigated the expression of BK channels in GBM stem‐like cells using electrophysiological and immunochemical techniques, and assessed their involvement in the migratory process of this important cell subpopulation. In U87‐MG cells, BK channel expression and function were markedly upregulated in GBM stem‐like cells expressing CD133, as well as other stem cell markers. In addition, transwell migration tests showed that GBM stem‐like cell migration was very sensitive to BK channel block.

  April 10, 2017   doi: 10.1002/jcp.25592   open full text
• TRIM33 is essential for osteoblast proliferation and differentiation via BMP pathway.
  Jia Guo, Wei Qin, Quan Xing, Manman Gao, Fuxin Wei, Zhi Song, Lingling Chen, Ying Lin, Xianling Gao, Zhengmei Lin.
  Journal of Cellular Physiology. April 10, 2017
  Tripartite motif containing 33 (TRIM33) functions both as a positive and negative regulator of the TGF‐β/BMP pathway in tumors; however, its effect and mechanism during osteoblast proliferation and differentiation, which involves the TGF‐β/BMP pathway is not defined. In this study, we used mouse C3H10T1/2 mesenchymal stem cell line and MC3T3‐E1 preosteoblasts to investigate the role of TRIM33 during this process. The results demonstrated that the expression of TRIM33 increased during the differentiation. Moreover, the overexpression or knockdown of TRIM33 resulted in both an augmentation or decrease in osteoblast differentiation, which were measured by the expression of alkaline phosphatase (ALP) at the mRNA level, both Runt‐related transcription factor 2 (Runx2) and osteocalcin (OCN) at the protein level, and the formation of mineral modules. To further demonstrate the mechanism of TRIM33 in this process, we found that TRIM33 could positively mediate the BMP pathway by forming TRIM33‐Smad1/5 complex. This interaction between TRIM33 and Smad1/5 triggered the phosphorylation of Smad1/5. In addition, the essential role of TRIM33 in osteoblast proliferation was determined in this study by CellCounting Kit (CCK) −8 and cell cycle assays. In summary, we establish the function of TRIM33 as a positive regulator of osteoblast differentiation in BMP pathway, which mediates its effect through its interaction with and activation of Smad1/5. In addition, the results clearly demonstrate that TRIM33 is necessary for osteoblast proliferation by regulating cell cycle. These results suggest that TRIM33 can be a positive target of osteoblast proliferation and differentiation through BMP pathway. We establish the function of TRIM33 as a positive regulator of osteoblast differentiation in BMP pathway, which mediates its effect through its interaction with and activation of Smad1/5. In addition, the results clearly demonstrate that TRIM33 is necessary for osteoblast proliferation by regulating cell cycle. These results suggest that TRIM33 can be a positive target of osteoblast proliferation and differentiation through BMP pathway.

  April 10, 2017   doi: 10.1002/jcp.25769   open full text
• Epithelial–mesenchymal transition (EMT): A biological process in the development, stem cell differentiation, and tumorigenesis.
  Tong Chen, Yanan You, Hua Jiang, Zack Z. Wang.
  Journal of Cellular Physiology. April 10, 2017
  The lineage transition between epithelium and mesenchyme is a process known as epithelial–mesenchymal transition (EMT), by which polarized epithelial cells lose their adhesion property and obtain mesenchymal cell phenotypes. EMT is a biological process that is often involved in embryogenesis and diseases, such as cancer invasion and metastasis. The EMT and the reverse process, mesenchymal–epithelial transition (MET), also play important roles in stem cell differentiation and de‐differentiation (or reprogramming). In this review, we will discuss current research progress of EMT in embryonic development, cellular differentiation and reprogramming, and cancer progression, all of which are representative models for researches of stem cell biology in normal and in diseases. Understanding of EMT and MET may help to identify specific markers to distinguish normal stem cells from cancer stem cells in future. In this review, we discuss current research progress of EMT in embryonic development, cellular differentiation and reprogramming, and cancer progression, all of which are representative models for researches of stem cell biology in normals and in diseases.

  April 10, 2017   doi: 10.1002/jcp.25797   open full text
• Neurotransmitters: The Critical Modulators Regulating Gut–Brain Axis.
  Rahul Mittal, Luca H. Debs, Amit P. Patel, Desiree Nguyen, Kunal Patel, Gregory O'Connor, M'hamed Grati, Jeenu Mittal, Denise Yan, Adrien A. Eshraghi, Sapna K. Deo, Sylvia Daunert, Xue Zhong Liu.
  Journal of Cellular Physiology. April 10, 2017
  Neurotransmitters, including catecholamines and serotonin, play a crucial role in maintaining homeostasis in the human body. Studies on these neurotransmitters mainly revolved around their role in the “fight or flight” response, transmitting signals across a chemical synapse and modulating blood flow throughout the body. However, recent research has demonstrated that neurotransmitters can play a significant role in the gastrointestinal (GI) physiology. Norepinephrine (NE), epinephrine (E), dopamine (DA), and serotonin have recently been a topic of interest because of their roles in the gut physiology and their potential roles in GI and central nervous system pathophysiology. These neurotransmitters are able to regulate and control not only blood flow, but also affect gut motility, nutrient absorption, GI innate immune system, and the microbiome. Furthermore, in pathological states, such as inflammatory bowel disease (IBD) and Parkinson's disease, the levels of these neurotransmitters are dysregulated, therefore causing a variety of GI symptoms. Research in this field has shown that exogenous manipulation of catecholamine serum concentrations can help in decreasing symptomology and/or disease progression. In this review article, we discuss the current state‐of‐the‐art research and literature regarding the role of neurotransmitters in regulation of normal GI physiology, their impact on several disease processes, and novel work focused on the use of exogenous hormones and/or psychotropic medications to improve disease symptomology. J. Cell. Physiol. 232: 2359–2372, 2017. © 2016 Wiley Periodicals, Inc. Neurotransmitters affect microbiota in the gut: Neurotransmitters, including serotonin, alter the microbiota in the gut that can modulate the production of cytokines and bacterial by‐products, leading to either healthy or diseased state.

  April 10, 2017   doi: 10.1002/jcp.25518   open full text
• MAP Kinase‐Dependent RUNX2 Phosphorylation Is Necessary for Epigenetic Modification of Chromatin During Osteoblast Differentiation.
  Yan Li, Chunxi Ge, Renny T. Franceschi.
  Journal of Cellular Physiology. April 10, 2017
  RUNX2, an essential transcription factor for osteoblast differentiation and bone formation is activated by ERK/MAP kinase‐dependent phosphorylation. However, relationship between these early events and specific epigenetic modifications of chromatin during osteoblast differentiation have not been previously examined. Here, we explore these relationships using chromatin immunoprecipitation (ChIP) to detect chromatin modifications in RUNX2‐binding regions of Bglap2 and Ibsp. Growth of MC3T3‐E1c4 preosteoblast cells in differentiation conditions rapidly induced Bglap2 and lbsp mRNAs. For both genes, osteogenic stimulation increased chromatin‐bound P‐ERK, P‐RUNX2, p300, and RNA polymerase II as well as histone H3K9 and H4K5 acetylation. The level of H3K4 di‐methylation, another gene activation‐associated histone mark, also increased. In contrast, levels of the gene repressive marks, H3K9 mono‐, di‐, and tri‐methylation in the same regions were reduced. Inhibition of MAP kinase signaling blocked differentiation‐dependent chromatin modifications and Bglap2 and Ibsp expression. To evaluate the role of RUNX2 phosphorylation in these responses, RUNX2‐deficient C3H10T1/2 cells were transduced with adenovirus encoding wild type or phosphorylation site mutant RUNX2 (RUNX2 S301A/S319A). Wild type RUNX2, but not the non‐phosphorylated mutant, increased H3K9 and H4K5 acetylation as well as chromatin‐associated P‐ERK, p300, and polymerase II. Thus, RUNX2 phosphorylation is necessary for subsequent epigenetic changes required for osteoblast gene expression. Taken together, this study reveals a molecular mechanism through which osteogenic genes are controlled by a MAPK and P‐RUNX2‐dependent process involving epigenetic modifications of specific promoter regions. J. Cell. Physiol. 232: 2427–2435, 2017. © 2016 Wiley Periodicals, Inc. The RUNX2 transcription factor requires MAP kinase‐dependent phosphorylation to become transcriptionally active. Here, we show that phosphorylation is required for transcription‐related changes in histone acetylation and methylation and recruitment of cofactors such as P300/CBP and RNA polymerase II to promoter regions of bone‐related genes.

  April 10, 2017   doi: 10.1002/jcp.25517   open full text
• Bone Shaft Revascularization After Marrow Ablation Is Dramatically Accelerated in BSP‐/‐ Mice, Along With Faster Hematopoietic Recolonization.
  Wafa Bouleftour, Renata Neves Granito, Arnaud Vanden‐Bossche, Odile Sabido, Bernard Roche, Mireille Thomas, Marie Thérèse Linossier, Jane E. Aubin, Marie‐Hélène Lafage‐Proust, Laurence Vico, Luc Malaval.
  Journal of Cellular Physiology. April 10, 2017
  The bone organ integrates the activity of bone tissue, bone marrow, and blood vessels and the factors ensuring this coordination remain ill defined. Bone sialoprotein (BSP) is with osteopontin (OPN) a member of the small integrin binding ligand N‐linked glycoprotein (SIBLING) family, involved in bone formation, hematopoiesis and angiogenesis. In rodents, bone marrow ablation induces a rapid formation of medullary bone which peaks by ∼8 days (d8) and is blunted in BSP‐/‐ mice. We investigated the coordinate hematopoietic and vascular recolonization of the bone shaft after marrow ablation of 2 month old BSP+/+ and BSP‐/‐ mice. At d3, the ablated area in BSP‐/‐ femurs showed higher vessel density (×4) and vascular volume (×7) than BSP+/+. Vessel numbers in the shaft of ablated BSP+/+ mice reached BSP‐/‐ values only by d8, but with a vascular volume which was twice the value in BSP‐/‐, reflecting smaller vessel size in ablated mutants. At d6, a much higher number of Lin− (×3) as well as LSK (Lin− IL‐7Rα− Sca‐1hi c‐Kithi, ×2) and hematopoietic stem cells (HSC: Flt3− LSK, ×2) were counted in BSP‐/‐ marrow, indicating a faster recolonization. However, the proportion of LSK and HSC within the Lin− was lower in BSP‐/‐ and more differentiated stages were more abundant, as also observed in unablated bone, suggesting that hematopoietic differentiation is favored in the absence of BSP. Interestingly, unablated BSP‐/‐ femur marrow also contains more blood vessels than BSP+/+, and in both intact and ablated shafts expression of VEGF and OPN are higher, and DMP1 lower in the mutants. In conclusion, bone marrow ablation in BSP‐/‐ mice is followed by a faster vascular and hematopoietic recolonization, along with lower medullary bone formation. Thus, lack of BSP affects the interplay between hematopoiesis, angiogenesis, and osteogenesis, maybe in part through higher expression of VEGF and the angiogenic SIBLING, OPN. J. Cell. Physiol. 232: 2528–2537, 2017. © 2016 Wiley Periodicals, Inc. In mice with a knockout of the SIBLING gene, Bone Sialoprotein (BSP‐/‐) vascular and hematopoietic recolonization are faster than in wild type, despite lower medullary bone formation. This goes with enhanced expression of osteopontin and VEGF, and downregulation of Dentin Matrix Protein 1, another SIBLING shown to inhibit angiogenesis. Lack of BSP thus affects the interplay between hematopoiesis, angiogenesis, and osteogenesis, in part through the mediation of pro‐ and antiangiogenic factors.

  April 10, 2017   doi: 10.1002/jcp.25630   open full text
• Use of a Collagen Membrane to Enhance the Survival of Primary Intestinal Epithelial Cells.
  Fiorella Di Claudio, Cecilia I. Muglia, Paola L. Smaldini, María Lucía Orsini Delgado, Fernando M. Trejo, J. Raúl Grigera, Guillermo H. Docena.
  Journal of Cellular Physiology. April 10, 2017
  Intestinal epithelial cell culture is important for biological, functional, and immunological studies. Since enterocytes have a short in vivo life span due to anoikis, we aimed to establish a novel and reproducible method to prolong the survival of mouse and human cells. Cells were isolated following a standard procedure, and cultured on ordered‐cow's collagen membranes. A prolonged cell life span was achieved; cells covered the complete surface of bio‐membranes and showed a classical enterocyte morphology with high expression of enzymes supporting the possibility of cryopreservation. Apoptosis was dramatically reduced and cultured enterocytes expressed cytokeratin and LGR5 (low frequency). Cells exposed to LPS or flagellin showed the induction of TLR4 and TLR5 expression and a functional phenotype upon exposure to the probiotic Bifidobacterium bifidum or the pathogenic Clostridium difficile. The secretion of the homeostatic (IL‐25 and TSLP), inhibitory (IL‐10 and TGF‐β), or pro‐inflammatory mediators (IL‐1β and TNF) were induced. In conclusion, this novel protocol using cow's collagen‐ordered membrane provides a simple and reproducible method to maintain intestinal epithelial cells functional for cell‐microorganism interaction studies and stem cell expansion. J. Cell. Physiol. 232: 2489–2496, 2017. © 2016 Wiley Periodicals, Inc. We have developed a method for culturing primary intestinal epithelial cells useful for the study of their interaction with microbes.

  April 10, 2017   doi: 10.1002/jcp.25594   open full text
• Cre Recombinase Strains Used for the Study of Adipose Tissues and Adipocyte Progenitors.
  Jiaqi Liu, Ziye Xu, Weiche Wu, Yizhen Wang, Tizhong Shan.
  Journal of Cellular Physiology. April 10, 2017
  Adipose tissues play important roles in whole body energy homeostasis and lifer span. Understanding the mechanisms of controlling adipose tissues development is significant for providing useful information to treat the worldwide epidemic of obesity and its associated metabolic diseases. Several different Cre transgenes have been generated and used for determining the origin of adipose tissues and the function of individual gene in regulating adipose growth and development. Here, we mainly review and discuss the efficiency and specific of those Cre recombinase mouse strains used for the study of adipose tissues and adipocyte progenitors. J. Cell. Physiol. 232: 2698–2703, 2017. © 2016 Wiley Periodicals, Inc. We mainly review and discuss the efficiency and specific of those Cre recombinase mouse strains used for determining the origin of adipose tissues and the function of individual gene in regulating adipose growth and development.

  April 10, 2017   doi: 10.1002/jcp.25675   open full text
• HSF1 phosphorylation by ERK/GSK3 suppresses RNF126 to sustain IGF‐IIR expression for hypertension‐induced cardiomyocyte hypertrophy.
  Chih‐Yang Huang, Fa‐Lun Lee, Shu‐Fen Peng, Kuan‐Ho Lin, Ray‐Jade Chen, Tsung‐Jung Ho, Fu‐Jen Tsai, V. Vijaya Padma, Wei‐Wen Kuo, Chih‐Yang Huang.
  Journal of Cellular Physiology. April 06, 2017
  Hypertension‐induced cardiac hypertrophy and apoptosis are major characteristics of early‐stage heart failure (HF). Inhibition of extracellular signal‐regulated kinases (ERK) efficaciously suppressed angiotensin II (ANG II)‐induced cardiomyocyte hypertrophy and apoptosis by blocking insulin‐like growth factor II receptor (IGF‐IIR) signaling. However, the detailed mechanism by which ANG II induces ERK‐mediated IGF‐IIR signaling remains elusive. Here, we found that ANG II activated ERK to upregulate IGF‐IIR expression via the angiotensin II type I receptor (AT1R). ERK activation subsequently phosphorylates HSF1 at serine 307, leading to a secondary phosphorylation by glycogen synthase kinase III (GSK3) at serine 303. Moreover, we found that ANG II mediated ERK/GSK3‐induced IGF‐IIR protein stability by downregulating the E3 ubiquitin ligase of IGF‐IIR RING finger protein CXXVI (RNF126). The expression of RNF126 decreased following ANG II‐induced HSF1S303 phosphorylation, resulting in IGF‐IIR protein stability and increased cardiomyocyte injury. Inhibition of GSK3 significantly alleviated ANG II‐induced cardiac hypertrophy in vivo and in vitro. Taken together, these results suggest that HSF1 phosphorylation stabilizes IGF‐IIR protein stability by downregulating RNF126 during cardiac hypertrophy. ANG II activates ERK/GSK3 to phosphorylate HSF1, resulting in RNF126 degradation, which stabilizes IGF‐IIR protein expression and eventually results in cardiac hypertrophy. HSF1 could be a valuable therapeutic target for cardiac diseases among hypertensive patients. This article is protected by copyright. All rights reserved

  April 06, 2017   doi: 10.1002/jcp.25945   open full text
• Maintenance of Bone Homeostasis by DLL1‐Mediated Notch Signaling.
  Yukari Muguruma, Katsuto Hozumi, Hiroyuki Warita, Takashi Yahata, Tomoko Uno, Mamoru Ito, Kiyoshi Ando.
  Journal of Cellular Physiology. March 31, 2017
  Adult bone mass is maintained through a balance of the activities of osteoblasts and osteoclasts. Although Notch signaling has been shown to maintain bone homeostasis by controlling the commitment, differentiation, and function of cells in both the osteoblast and osteoclast lineages, the precise mechanisms by which Notch performs such diverse and complex roles in bone physiology remain unclear. By using a transgenic approach that modified the expression of delta‐like 1 (DLL1) or Jagged1 (JAG1) in an osteoblast‐specific manner, we investigated the ligand‐specific effects of Notch signaling in bone homeostasis. This study demonstrated for the first time that the proper regulation of DLL1 expression, but not JAG1 expression, in osteoblasts is essential for the maintenance of bone remodeling. DLL1‐induced Notch signaling was responsible for the expansion of the bone‐forming cell pool by promoting the proliferation of committed but immature osteoblasts. However, DLL1‐Notch signaling inhibited further differentiation of the expanded osteoblasts to become fully matured functional osteoblasts, thereby substantially decreasing bone formation. Osteoblast‐specific expression of DLL1 did not alter the intrinsic differentiation ability of cells of the osteoclast lineage. However, maturational arrest of osteoblasts caused by the DLL1 transgene impaired the maturation and function of osteoclasts due to a failed osteoblast‐osteoclast coupling, resulting in severe suppression of bone metabolic turnover. Taken together, DLL1‐mediated Notch signaling is critical for proper bone remodeling as it regulates the differentiation and function of both osteoblasts and osteoclasts. Our study elucidates the importance of ligand‐specific activation of Notch signaling in the maintenance of bone homeostasis. J. Cell. Physiol. 232: 2569–2580, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc. The proper regulation of DLL1 expression, but not JAG1 expression, in osteoblasts is essential for the maintenance of bone remodeling. DLL1‐Notch signaling directly controls the physiological expansion and differentiation of osteoblasts and indirectly affects the maturation and function of osteoclasts, thereby ensuring the homeostasis of bone mass.

  March 31, 2017   doi: 10.1002/jcp.25647   open full text
• Reduced primary cilia length and altered Arl13b expression are associated with deregulated chondrocyte Hedgehog signaling in alkaptonuria.
  Stephen D. Thorpe, Silvia Gambassi, Clare L. Thompson, Charmilie Chandrakumar, Annalisa Santucci, Martin M. Knight.
  Journal of Cellular Physiology. March 31, 2017
  Alkaptonuria (AKU) is a rare inherited disease resulting from a deficiency of the enzyme homogentisate 1,2‐dioxygenase which leads to the accumulation of homogentisic acid (HGA). AKU is characterized by severe cartilage degeneration, similar to that observed in osteoarthritis. Previous studies suggest that AKU is associated with alterations in cytoskeletal organization which could modulate primary cilia structure/function. This study investigated whether AKU is associated with changes in chondrocyte primary cilia and associated Hedgehog signaling which mediates cartilage degradation in osteoarthritis. Human articular chondrocytes were obtained from healthy and AKU donors. Additionally, healthy chondrocytes were treated with HGA to replicate AKU pathology (+HGA). Diseased cells exhibited shorter cilia with length reductions of 36% and 16% in AKU and +HGA chondrocytes respectively, when compared to healthy controls. Both AKU and +HGA chondrocytes demonstrated disruption of the usual cilia length regulation by actin contractility. Furthermore, the proportion of cilia with axoneme breaks and bulbous tips was increased in AKU chondrocytes consistent with defective regulation of ciliary trafficking. Distribution of the Hedgehog‐related protein Arl13b along the ciliary axoneme was altered such that its localization was increased at the distal tip in AKU and +HGA chondrocytes. These changes in cilia structure/trafficking in AKU and +HGA chondrocytes were associated with a complete inability to activate Hedgehog signaling in response to exogenous ligand. Thus, we suggest that altered responsiveness to Hedgehog, as a consequence of cilia dysfunction, may be a contributing factor in the development of arthropathy highlighting the cilium as a novel target in AKU. This study investigates whether the rare inherited disease alkaptonuria (AKU) is associated with changes in chondrocyte primary cilia and associated Hedgehog signaling, which is known to mediate cartilage degradation in osteoarthritis. AKU results in alterations to primary cilia structure which are associated with ciliary trafficking defects with the potential to impact a range of fundamental signaling pathways. In particular, AKU chondrocytes demonstrated a complete inability to activate Hedgehog signaling in response to endogenous ligand; which we propose may be the result of the observed dysfunction of cilia structure and trafficking. Thus, we suggest that altered responsiveness to Hedgehog, as a consequence of cilia dysfunction, may be a contributing factor in the development of arthropathy highlighting the cilium as a novel target in AKU.

  March 31, 2017   doi: 10.1002/jcp.25839   open full text
• Crosstalk of autophagy and apoptosis: Involvement of the dual role of autophagy under ER stress.
  Shuling Song, Jin Tan, Yuyang Miao, Mengmeng Li, Qiang Zhang.
  Journal of Cellular Physiology. March 31, 2017
  Endoplasmic reticulum (ER) stress is a common cellular stress response that is triggered by a variety of conditions that disturb cellular homeostasis, and induces cell apoptosis. Autophagy, an important and evolutionarily conserved mechanism for maintaining cellular homeostasis, is closely related to the apoptosis induced by ER stress. There are common upstream signaling pathways between autophagy and apoptosis induced by ER stress, including PERK/ATF4, IRE1α, ATF6, and Ca2+. Autophagy can not only block the induction of apoptosis by inhibiting the activation of apoptosis‐associated caspase which could reduce cellular injury, but also help to induce apoptosis. In addition, the activation of apoptosis‐related proteins can also inhibit autophagy by degrading autophagy‐related proteins, such as Beclin‐1, Atg4D, Atg3, and Atg5. Although the interactions of different autophagy‐ and apoptosis‐related proteins, and also common upstream signaling pathways have been found, the potential regulatory mechanisms have not been clearly understood. In this review, we summarize the dual role of autophagy, and the interplay and potential regulatory mechanisms between autophagy and apoptosis under ER stress condition. The dura role of autophagy, pro‐survival, and ‐death, may be dependent on the extent of ER stress, and it implements by the regulation of apoptosis. Autophagy can not only block the induction of apoptosis by inhibiting the activation of apoptosis‐associated caspase which could reduce cellular injury, but also help to induce apoptosis. In addition, the activation of apoptosis‐related proteins can also inhibit autophagy by degrading autophagy‐related proteins, such as Beclin‐1, Atg4D, Atg3, and Atg5.

  March 31, 2017   doi: 10.1002/jcp.25785   open full text
• Staphylococcal protein A promotes osteoclastogenesis through MAPK signaling during bone infection.
  Yuan Wang, Xin Liu, Ce Dou, Zhen Cao, Chuan Liu, Shiwu Dong, Jun Fei.
  Journal of Cellular Physiology. March 31, 2017
  Bone infection is a common and serious complication in the orthopedics field, which often leads to excessive bone destruction and non‐union. Osteoclast is the only type of cells which have the function of bone resorption. Its over activation is closely related to excessive bone loss. Staphylococcus aureus (S. aureus) is a major pathogen causing bone infection, which can produce a large number of strong pathogenic substances staphylococcal protein A (SPA). However, few studies were reported about the effects of SPA on osteoclastogenesis. In our study, we observed that S. aureus activated osteoclasts and promoted bone loss in bone infection specimens. Then, we investigated the effects of SPA on RANKL‐induced osteoclastogenesis in vitro, the results revealed that SPA promoted osteoclastic differentiation and fusion, and enhanced osteoclastic bone resorption. In addition, we also showed that SPA upregulated the expression of NFATc1 and c‐FOS through the activation of MAPK signaling to promote osteoclastogenesis. Our findings might help us better understand the pathogenic role of S. aureus in bone infection and develop new therapeutic strategies for infectious bone diseases. Staphylococcal protein A (SPA) is one of the most important pathogenic components of Staphylococcus aureus (S. aureus). Here we reported that S. aureus caused bone loss in bone infection specimens. In vitro study showed that SPA promoted osteoclastic differentiation, fusion and bone resorption activity by upregulating NFATc1 and c‐FOS through MAPK signaling.

  March 31, 2017   doi: 10.1002/jcp.25774   open full text
• Challenging of AS160/TBC1D4 Alters Intracellular Lipid milieu in L6 Myotubes Incubated With Palmitate.
  Agnieszka Mikłosz, Bartłomiej Łukaszuk, Małgorzata Żendzian‐Piotrowska, Justyna Brańska‐Januszewska, Halina Ostrowska, Adrian Chabowski.
  Journal of Cellular Physiology. March 31, 2017
  The Akt substrate of 160 kDa (AS160) is a key regulator of GLUT4 translocation from intracellular depots to the plasma membrane in myocytes. Likely, AS160 also controls LCFAs transport, which requires relocation of fatty acid transporters. The aim of the present study was to determine the impact of AS160 knockdown on lipid milieu in L6 myotubes incubated with palmitate (PA). Therefore, we compared two different settings, namely: 1) AS160 knockdown prior to palmitate incubation (pre‐PA‐silencing, AS160−/PA); 2) palmitate incubation with subsequent AS160 knockdown (post‐PA‐silencing, PA/AS160−). The efficiency of AS160 silencing was checked at mRNA and protein levels. The expression and localization of FA transporters were determined using Western Blot and immunofluorescence analyses. Intracellular lipid content (FFA, DAG, TAG, and PL) and FA composition were estimated by GLC, whereas basal palmitate uptake was analyzed by means of scintigraphy. Both groups with silenced AS160 were characterized by a greater expression of FA transporters (FAT/CD36, FATP‐1, 4) which had contributed to an increased FA cellular influx. Accordingly, we observed that post‐PA‐silencing of AS160 resulted in a marked decrement in DAG, TAG, and PL contents, but increased FFA content (PA/AS160− vs. PA). The opposite effect was observed in the group with pre‐PA‐silencing of AS160 in which AS160 knockdown did not affect the lipid pools (AS160−/PA vs. PA). Our results indicate that post‐PA‐silencing of AS160 has a capacity to decrease the lipotoxic effect(s) of PA by decreasing the content of lipids (DAG and PL) that promote insulin resistance in myotubes. J. Cell. Physiol. 232: 2373–2386, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals Inc. AS160/TBC1D4 is a key regulator of GLUT4 translocation to the plasma membrane in myocytes, moreover, it role in the control of protein mediated fatty acids transport is also likely. The current study indicates that AS160/TBC1D4 may influence not only FA transporters content, and therefore their influx, but also their storage and oxidation. However, the effect differs depending on the events sequence, i.e., AS160 knock‐down prior to or after palmitate incubation.

  March 31, 2017   doi: 10.1002/jcp.25632   open full text
• Pyk2 inhibition promotes contractile differentiation in arterial smooth muscle.
  Mario Grossi, Anirban Bhattachariya, Ina Nordström, Karolina M. Turczyńska, Daniel Svensson, Sebastian Albinsson, Bengt‐Olof Nilsson, Per Hellstrand.
  Journal of Cellular Physiology. March 31, 2017
  Modulation from contractile to synthetic phenotype of vascular smooth muscle cells is a central process in disorders involving compromised integrity of the vascular wall. Phenotype modulation has been shown to include transition from voltage‐dependent toward voltage‐independent regulation of the intracellular calcium level, and inhibition of non‐voltage dependent calcium influx contributes to maintenance of the contractile phenotype. One possible mediator of calcium‐dependent signaling is the FAK‐family non‐receptor protein kinase Pyk2, which is activated by a number of stimuli in a calcium‐dependent manner. We used the Pyk2 inhibitor PF‐4594755 and Pyk2 siRNA to investigate the role of Pyk2 in phenotype modulation in rat carotid artery smooth muscle cells and in cultured intact arteries. Pyk2 inhibition promoted the expression of smooth muscle markers at the mRNA and protein levels under stimulation by FBS or PDGF‐BB and counteracted phenotype shift in cultured intact carotid arteries and balloon injury ex vivo. During long‐term (24–96 hr) treatment with PF‐4594755, smooth muscle markers increased before cell proliferation was inhibited, correlating with decreased KLF4 expression and differing from effects of MEK inhibition. The Pyk2 inhibitor reduced Orai1 and preserved SERCA2a expression in carotid artery segments in organ culture, and eliminated the inhibitory effect of PDGF stimulation on L‐type calcium channel and large‐conductance calcium‐activated potassium channel expression in carotid cells. Basal intracellular calcium level, calcium wave activity, and store‐operated calcium influx were reduced after Pyk2 inhibition of growth‐stimulated cells. Pyk2 inhibition may provide an interesting approach for preserving vascular smooth muscle differentiation under pathophysiological conditions. The Pyk2 inhibitor PF‐4594755 potently inhibits Pyk2 phosphorylation of vascular smooth muscle cells stimulated by FBS or PDGF‐BB. This is accompanied by increased expression of smooth muscle marker proteins without concomitant effects on cell proliferation. Pyk2 inhibition upregulates SERCA2a expression and may preserve the contractile phenotype by lowering basal intracellular calcium and promoting voltage‐dependent control of calcium influx over the plasma membrane.

  March 31, 2017   doi: 10.1002/jcp.25760   open full text
• Mactosylceramide prevents glial cell overgrowth by inhibiting insulin and fibroblast growth factor receptor signaling.
  Stine Gerdøe‐Kristensen, Viktor K. Lund, Hans H. Wandall, Ole Kjaerulff.
  Journal of Cellular Physiology. March 31, 2017
  Receptor tyrosine kinase (RTK) signaling controls key aspects of cellular differentiation, proliferation, survival, metabolism, and migration. Deregulated RTK signaling also underlies many cancers. Glycosphingolipids (GSL) are essential elements of the plasma membrane. By affecting clustering and activity of membrane receptors, GSL modulate signal transduction, including that mediated by the RTK. GSL are abundant in the nervous system, and glial development in Drosophila is emerging as a useful model for studying how GSL modulate RTK signaling. Drosophila has a simple GSL biosynthetic pathway, in which the mannosyltransferase Egghead controls conversion of glucosylceramide (GlcCer) to mactosylceramide (MacCer). Lack of elongated GSL in egghead (egh) mutants causes overgrowth of subperineurial glia (SPG), largely due to aberrant activation of phosphatidylinositol 3‐kinase (PI3K). However, to what extent this effect involves changes in upstream signaling events is unresolved. We show here that glial overgrowth in egh is strongly linked to increased activation of Insulin and fibroblast growth factor receptors (FGFR). Glial hypertrophy is phenocopied when overexpressing gain‐of‐function mutants of the Drosophila insulin receptor (InR) and the FGFR homolog Heartless (Htl) in wild type SPG, and is suppressed by inhibiting Htl and InR activity in egh. Knockdown of GlcCer synthase in the SPG fails to suppress glial overgrowth in egh nerves, and slightly promotes overgrowth in wild type, suggesting that RTK hyperactivation is caused by absence of MacCer and not by GlcCer accumulation. We conclude that an early product in GSL biosynthesis, MacCer, prevents inappropriate activation of insulin and fibroblast growth factor receptors in Drosophila glia. Receptor tyrosine kinase signaling controls key aspects of cellular differentiation, proliferation, survival, metabolism, and migration. Glycosphingolipids modulate signal transduction, including that mediated by the receptor tyrosine kinases. We report that an early product in glycosphingolipid biosynthesis, mactosylceramide, prevents inappropriate activation of insulin and fibroblast growth factor receptors in Drosophila glial cells.

  March 31, 2017   doi: 10.1002/jcp.25762   open full text
• Acetoacetate induces hepatocytes apoptosis by the ROS‐mediated MAPKs pathway in ketotic cows.
  Xiliang Du, Zhen Shi, Zhicheng Peng, Chenxu Zhao, Yuming Zhang, Zhe Wang, Xiaobing Li, Guowen Liu, Xinwei Li.
  Journal of Cellular Physiology. March 31, 2017
  Dairy cows with ketosis are characterized by oxidative stress, hepatic damage, and hyperketonemia. Acetoacetate (AA) is the main component of ketone bodies in ketotic cows, and is associated with the above pathological process. However, the potential mechanism was not illuminated. Therefore, the aim of this study was to investigate the mechanism of AA‐induced hepatic oxidative damage in ketotic cows. Compared with healthy cows, ketotic cows exhibited severe oxidative stress and hepatic damage. Moreover, the extent of hepatic damage and oxidative stress had a positive relationship with the AA levels. In vitro, AA treatment increased reactive oxygen species (ROS) content and further induced oxidative stress and apoptosis of bovine hepatocytes. In this process, AA treatment increased the phosphorylation levels of JNK and p38MAPK and decreased the phosphorylation level of ERK, which could increase p53 and inhibit nuclear factor E2‐related factor 2 (Nrf2) expression, nuclear localization, and DNA‐binding affinity, thereby inducing the overexpression of pro‐apoptotic molecules Bax, Caspase 3, Caspase 9, PARP and inhibition of anti‐apoptotic molecule Bcl‐2. Antioxidant N‐acetylcysteine (NAC) treatment or interference of MAPKs pathway could attenuate the hepatocytes apoptosis induced by AA. Collectively, these results indicate that AA triggers hepatocytes apoptosis via the ROS‐mediated MAPKs pathway in ketotic cows. Excess acetoacetate induced imbalance of oxidation and antioxidation, resulting in overproduction of reactive oxygen species (ROS) in the mitochondria. High concentration of ROS increased the phosphorylation levels of p38MAPK and JNK and decreased the phosphorylation levels of ERK1/2, which could activate p53 and inhibit Nrf2 expression, nuclear localization, and DNA‐binding affinity. Consequently, the expression of pro‐apoptotic molecules Bax, Casepase9, Casepase3, and PARP was significantly upregulated and anti‐apoptotic molecules Bcl‐2 were markedly downregulated, resulting in hepatocytes apoptosis in ketotic cows.

  March 31, 2017   doi: 10.1002/jcp.25773   open full text
• Biological and pharmacological evaluation of dimethoxycurcumin: A metabolically stable curcumin analogue with a promising therapeutic potential.
  Manouchehr Teymouri, Nastaran Barati, Matteo Pirro, Amirhosein Sahebkar.
  Journal of Cellular Physiology. March 31, 2017
  Dimethoxycurcumin (DiMC) is a synthetic analog of curcumin with superior inter‐related pro‐oxidant and anti‐cancer activity, and metabolic stability. Numerous studies have shown that DiMC reserves the biologically beneficial features, including anti‐inflammatory, anti‐carcinogenic, and cytoprotective properties, almost to the same extent as curcumin exhibits. DiMC lacks the phenolic‐OH groups as opposed to curcumin, dimethoxycurcumin, and bis‐demethoxycurcumin that all vary in the number of methoxy groups per molecule, and has drawn the attentions of researchers who attempted to discover the structure‐activity relationship (SAR) of curcumin. In this regard, tetrahydrocurcumin (THC), the reduced and biologically inert metabolite of curcumin, denotes the significance of the conjugated α,β diketone moiety for the curcumin activity. DiMC exerts unique molecular activities compared to curcumin, including induction of androgen receptor (AR) degradation and suppression of the transcription factor activator protein‐1 (AP‐1). The enhanced AR degradation on DiMC treatment suggests it as a novel anticancer agent against resistant tumors with androgenic etiology. Further, DiMC might be a potential treatment for acne vulgaris. DiMC induces epigenetic alteration more effectively than curcumin, although both showed no direct DNA hypomethylating activity. Given the metabolic stability, nanoparticulation of DiMC is more promising for in vivo effectiveness. However, studies in this regard are still in its infancy. In the current review, we portray the various molecular and biological functions of DiMC reported so far. Whenever possible, the efficiency is compared with curcumin and the reasons for DiMC being more metabolically stable are elaborated. We also provide future perspective investigations with respect to varying DiMC‐nanoparticles. This review, being the first of its kind, is a comprehensive evaluation of the molecular, cellular, and clinical properties of dimethoxycurcumin as a promising analog of curcumin. We have discussed various molecular and biological functions of DiMC reported so far. Whenever possible, the efficiency compared with curcumin and the reasons for DiMC being more metabolically stable than curcumin has been explained. We have also provided future perspective investigations with respect to varying DiMC‐nanoparticles.

  March 31, 2017   doi: 10.1002/jcp.25749   open full text
• Therapeutic effects of curcumin in inflammatory and immune‐mediated diseases: A nature‐made jack‐of‐all‐trades?
  Elham Abdollahi, Amir Abbas Momtazi, Thomas P. Johnston, Amirhossein Sahebkar.
  Journal of Cellular Physiology. March 31, 2017
  Curcumin is a dietary polyphenol from turmeric with numerous pharmacological activities. Novel animal and human studies indicate that curcumin can affect different immune cells, such as various T lymphocyte subsets, macrophages, dendritic cells, B lymphocytes and natural killer cells, which results in decreasing severity of various diseases with immunological etiology. The present review provides a comprehensive overview of the effects of curcumin on different immune cells and immune system‐related diseases. The present review provides a comprehensive overview of the effects of curcumin on different immune cells and immune system‐related diseases.

  March 31, 2017   doi: 10.1002/jcp.25778   open full text
• Cyclic‐glycine‐proline accelerates mammary involution by promoting apoptosis and inhibiting IGF‐1 function.
  Gagandeep Singh‐Mallah, Christopher D. McMahon, Jian Guan, Kuljeet Singh.
  Journal of Cellular Physiology. March 31, 2017
  In rodents, post‐lactational involution of mammary glands is characterized by the loss of mammary epithelial cells via apoptosis, which is associated with a decline in the expression of insulin‐like growth factor‐1 (IGF‐1). Overexpression of IGF‐1 delays involution by inhibiting apoptosis of epithelial cells and preserving the remaining secretory alveoli. Cyclic‐glycine‐proline (cGP), a metabolite of IGF‐1, normalizes IGF‐1 function under pathological conditions by regulating the bioavailability of IGF‐1. The present study investigated the effect of cGP on the physiological decline in IGF‐1 function during post‐lactational mammary involution. Rat dams were gavaged with either cGP (3 mg/kg) or saline once per day from post‐natal d8‐22. Before collecting tissue on post‐natal d23, a pair of mammary glands were sealed on d20 (72 hr‐engorgement, thus representative of late‐involution) and d22 (24 hr‐engorgement, thus representative of mid‐involution), while the remaining glands were allowed to involute naturally (early‐involution). During early‐involution, cGP accelerated the loss of mammary cells through apoptosis, resulting in an earlier clearance of intact secretory alveoli compared with the control group. This coincided with an earlier up‐regulation of the cell survival factors, Bcl‐xl and IGF‐1R, in the early‐involution cGP glands compared with the control glands. During late‐involution, cGP reduced the bioactivity of IGF‐1, which was evident through decreased phosphorylation of IGF‐1R in the regressed alveoli. Maternal administration of cGP did not alter milk production and composition during early‐, peak‐, or late‐stage of lactation. These data show that cGP accelerates post‐lactational involution by promoting apoptosis and the physiological decline in IGF‐1 function. Role of cGP in regulating IGF‐1 function during brain development and in neurological conditions is well known. This study shows that cGP accelerates the onset of post‐lactational involution in mammary glands by promoting the physiological decline in IGF‐1 function.

  March 31, 2017   doi: 10.1002/jcp.25782   open full text
• Effects of Carbocysteine and Beclomethasone on Histone Acetylation/Deacetylation Processes in Cigarette Smoke Exposed Bronchial Epithelial Cells.
  Elisabetta Pace, Serena Di Vincenzo, Maria Ferraro, Liboria Siena, Giuseppina Chiappara, Paola Dino, Patrizio Vitulo, Alessandro Bertani, Federico Saibene, Luigi Lanata, Mark Gjomarkaj.
  Journal of Cellular Physiology. March 31, 2017
  Histone deacetylase expression/activity may control inflammation, cell senescence, and responses to corticosteroids. Cigarette smoke exposure, increasing oxidative stress, may negatively affect deacetylase expression/activity. The effects of cigarette smoke extracts (CSE), carbocysteine, and beclomethasone dipropionate on chromatin remodeling processes in human bronchial epithelial cells are largely unknown. The present study was aimed to assess the effects of cigarette smoke, carbocysteine, and beclomethasone dipropionate on histone deacetylase 3 (HDAC3) expression/activity, N‐CoR (nuclear receptor corepressor) expression, histone acetyltransferases (HAT) (p300/CBP) expression, p‐CREB and IL‐1 m‐RNA expression, neutrophil chemotaxis. Increased p‐CREB expression was observed in the bronchial epithelium of smokers. CSE increased p‐CREB expression and decreased HDAC3 expression and activity and N‐CoR m‐RNA and protein expression. At the same time, CSE increased the expression of the HAT, p300/CBP. All these events increased acetylation processes within the cells and were associated to increased IL‐1 m‐RNA expression and neutrophil chemotaxis. The incubation of CSE exposed cells with carbocysteine and beclomethasone counteracted the effects of cigarette smoke on HDAC3 and N‐CoR but not on p300/CBP. The increased deacetylation processes due to carbocysteine and beclomethasone dipropionate incubation is associated to reduced p‐CREB, IL‐1 m‐RNA expression, neutrophil chemotaxis. These findings suggest a new role of combination therapy with carbocysteine and beclomethasone dipropionate in restoring deacetylation processes compromised by cigarette smoke exposure. J. Cell. Physiol. 232: 2851–2859, 2017. © 2016 Wiley Periodicals, Inc. Cigarette smoke exposure, increasing oxidative stress, may negatively affect deacetylase expression/activity, thus, supporting increased pro‐inflammatory responses.The findings here provided suggest a new role of combination therapy with carbocysteine and beclomethasone dipropionate in restoring deacetylation processes compromised by cigarette smoke exposure.

  March 31, 2017   doi: 10.1002/jcp.25710   open full text
• Smoothened‐antagonists reverse homogentisic acid‐induced alterations of Hedgehog signaling and primary cilium length in alkaptonuria.
  Silvia Gambassi, Michela Geminiani, Stephen D. Thorpe, Giulia Bernardini, Lia Millucci, Daniela Braconi, Maurizio Orlandini, Clare L. Thompson, Elena Petricci, Fabrizio Manetti, Maurizio Taddei, Martin M. Knight, Annalisa Santucci.
  Journal of Cellular Physiology. March 29, 2017
  Alkaptonuria (AKU) is an ultra‐rare genetic disease, in which the accumulation of a toxic metabolite, homogentisic acid (HGA) leads to the systemic development of ochronotic aggregates. These aggregates cause severe complications mainly at the level of joints with extensive degradation of the articular cartilage. Primary cilia have been demonstrated to play an essential role in development and the maintenance of articular cartilage homeostasis, through their involvement in mechanosignaling and Hedgehog signaling pathways. Hedgehog signaling has been demonstrated to be activated in osteoarthritis (OA) and to drive cartilage degeneration in vivo. The numerous similarities between OA and AKU suggest that primary cilia Hedgehog signaling may also be altered in AKU. Thus, we characterized an AKU cellular model in which healthy chondrocytes were treated with HGA (66 µM) to replicate AKU cartilage pathology. We investigated the degree of activation of the Hedgehog signaling pathway and how treatment with inhibitors of the receptor Smoothened (Smo) influenced Hedgehog activation and primary cilia structure. The results obtained in this work provide a further step in the comprehension of the pathophysiological features of AKU, suggesting a potential therapeutic approach to modulate AKU cartilage degradation processes through manipulation of the Hedgehog pathway. Primary cilia of alkaptonuric (AKU) chondrocytes are shorter as compared to healthy cells and HGA treatment leads to the same phenotype. Primary cilia alterations are correlated to an aberrant Hedgehog (Hh) signaling activation. Treatment of HGA‐treated chondrocytes with antagonists of Smoothened reported cilia lengths and Hh activation to the normal levels.

  March 29, 2017   doi: 10.1002/jcp.25761   open full text
• Signaling in the Auditory System: Implications in Hair Cell Regeneration and Hearing Function.
  Rahul Mittal, Luca H. Debs, Desiree Nguyen, Amit P. Patel, M'hamed Grati, Jeenu Mittal, Denise Yan, Adrien A. Eshraghi, Xue Zhong Liu.
  Journal of Cellular Physiology. March 29, 2017
  Ear is a sensitive organ involved in hearing and balance function. The complex signaling network in the auditory system plays a crucial role in maintaining normal physiological function of the ear. The inner ear comprises a variety of host signaling pathways working in synergy to deliver clear sensory messages. Any disruption, as minor as it can be, has the potential to affect this finely tuned system with temporary or permanent sequelae including vestibular deficits and hearing loss. Mutations linked to auditory symptoms, whether inherited or acquired, are being actively researched for ways to reverse, silence, or suppress them. In this article, we discuss recent advancements in understanding the pathways involved in auditory system signaling, from hair cell development through transmission to cortical centers. Our review discusses Notch and Wnt signaling, cell to cell communication through connexin and pannexin channels, and the detrimental effects of reactive oxygen species on the auditory system. There has been an increased interest in the auditory community to explore the signaling system in the ear for hair cell regeneration. Understanding signaling pathways in the auditory system will pave the way for the novel avenues to regenerate sensory hair cells and restore hearing function. J. Cell. Physiol. 232: 2710–2721, 2017. © 2016 Wiley Periodicals, Inc. Wnt signaling influences hair cell differentiation. The presence of Wnt causes progenitor cells to differentiate into hair cell. Without Wnt, the progenitor cells become supporting cells.

  March 29, 2017   doi: 10.1002/jcp.25695   open full text
• LIPUS suppressed LPS‐induced IL‐1α through the inhibition of NF‐κB nuclear translocation via AT1‐PLCβ pathway in MC3T3‐E1 cells.
  Mayu Nagao, Natsuko Tanabe, Soichiro Manaka, Masako Naito, Jumpei Sekino, Tadahiro Takayama, Takayuki Kawato, Go Torigoe, Shunichiro Kato, Naoya Tsukune, Masao Maeno, Naoto Suzuki, Shuichi Sato.
  Journal of Cellular Physiology. March 29, 2017
  Inflammatory cytokines, interleukin (IL)‐1, IL‐6, and TNF‐α, are involved in inflammatory bone diseases such as rheumatoid osteoarthritis and periodontal disease. Particularly, periodontal disease, which destroys alveolar bone, is stimulated by lipopolysaccharide (LPS). Low‐intensity pulsed ultrasound (LIPUS) is used for bone healing in orthopedics and dental treatments. However, the mechanism underlying effects of LIPUS on LPS‐induced inflammatory cytokine are not well understood. We therefore aimed to investigate the role of LIPUS on LPS‐induced IL‐1α production. Mouse calvaria osteoblast‐like cells MC3T3‐E1 were incubated in the presence or absence of LPS (Porphyromonas gingivalis), and then stimulated with LIPUS for 30 min/day. To investigate the role of LIPUS, we determined the expression of IL‐1α stimulated with LIPUS and treated with an angiotensin II receptor type 1 (AT1) antagonist, Losartan. We also investigate to clarify the pathway of LIPUS, we transfected siRNA silencing AT1 (siAT1) in MC3T3‐E1. LIPUS inhibited mRNA and protein expression of LPS‐induced IL‐1α. LIPUS also reduced the nuclear translocation of NF‐κB by LPS‐induced IL‐1α. Losartan and siAT1 blocked all the stimulatory effects of LIPUS on IL‐1α production and IL‐1α‐mediated NF‐κB translocation induced by LPS. Furthermore, PLCβ inhibitor U73122 recovered NF‐κB translocation. These results suggest that LIPUS inhibits LPS‐induced IL‐1α via AT1‐PLCβ in osteoblasts. We exhibit that these findings are in part of the signaling pathway of LIPUS on the anti‐inflammatory effects of IL‐1α expression. LIPUS reduced IL‐1α production induced by LPS. LPS increased NF‐κB nuclear translocation. LIPUS stimulates AT1 receptor that acts inhibition of NF‐κB nuclear translocation.

  March 29, 2017   doi: 10.1002/jcp.25777   open full text
• Production of Macrophage Inhibitory Factor (MIF) by Primary Sertoli Cells; Its Possible Involvement in Migration of Spermatogonial Cells.
  Mahmoud Huleihel, Maram Abofoul‐Azab, Yael Abarbanel, Iris Einav, Elyahu Levitas, Eitan Lunenfeld.
  Journal of Cellular Physiology. March 29, 2017
  Macrophage migration inhibitory factor (MIF) is a multifunctional molecule. MIF was originally identified as a T‐cell‐derived factor responsible for the inhibition of macrophage migration. In testicular tissue of adult rats, MIF is constitutively expressed by Leydig cells under physiological conditions. The aim of this study was to examine MIF levels in testicular homogenates from different aged mice, and the capacity of Sertoli cells to produce it. We also examined MIF involvement in spermatogonial cell migration. Similar levels of MIF protein were detected in testicular homogenates of mice of different ages (1–8‐week‐old). However, the RNA expression levels of MIF were high in 1‐week‐old mice and significantly decreased with age compared to 1‐week‐old mice. MIF was stained in Sertoli, Leydig cells, and developed germ cells in the seminiferous tubules. Isolated Sertoli cells from 1‐week‐old mice stained to MIF. Cultures of Sertoli cells from 1‐week‐old mice produced and expressed high levels of MIF which significantly decreased with age. MIF was localized in the cytoplasm and nucleus of Sertoli cell cultures isolated from 1‐week‐old mice; however, it was localized only in the cytoplasm and branches of cultures isolated from 8‐week‐old mice. MIFR was detected in GFRα1 and Sertoli cells. MIF could induce migration of spermatogonial cells, and this effect was synergistic with glial cell‐line neurotrophic factor. Our results show, for the first time, the capacity of Sertoli cells to produce MIF under normal conditions and that MIFR expressed in GFRα1 and Sertoli cells. We also showed that MIF induced spermatogonial cell migration. J. Cell. Physiol. 232: 2869–2877, 2017. © 2016 Wiley Periodicals, Inc. Our study showed that Sertoli cells and spermatogonial cells generate MIF and express MIF receptor. MIF could induce spermatogonial cell migration.

  March 29, 2017   doi: 10.1002/jcp.25718   open full text
• Epigenetic Signatures at the RUNX2‐P1 and Sp7 Gene Promoters Control Osteogenic Lineage Commitment of Umbilical Cord‐Derived Mesenchymal Stem Cells.
  Hugo Sepulveda, Rodrigo Aguilar, Catalina P. Prieto, Francisco Bustos, Sócrates Aedo, José Lattus, Brigitte van Zundert, Veronica Palma, Martin Montecino.
  Journal of Cellular Physiology. March 28, 2017
  Wharton's Jelly mesenchymal stem cells (WJ‐MSCs) are an attractive potential source of multipotent stem cells for bone tissue replacement therapies. However, the molecular mechanisms involved in their osteogenic conversion are poorly understood. Particularly, epigenetic control operating at the promoter regions of the two master regulators of the osteogenic program, RUNX2/P57 and SP7 has not yet been described in WJ‐MSCs. Via quantitative PCR profiling and chromatin immunoprecipitation (ChIP) studies, here we analyze the ability of WJ‐MSCs to engage osteoblast lineage. In undifferentiated WJ‐MSCs, RUNX2/P57 P1, and SP7 promoters are found deprived of significant levels of the histone post‐translational marks that are normally associated with transcriptionally active genes (H3ac, H3K27ac, and H3K4me3). Moreover, the RUNX2 P1 promoter lacks two relevant histone repressive marks (H3K9me3 and H3K27me3). Importantly, RUNX2 P1 promoter is found highly enriched in the H3K4me1 mark, which has been shown recently to mediate gene repression of key regulatory genes. Upon induction of WJ‐MSCs osteogenic differentiation, we found that RUNX2/P57, but not SP7 gene expression is strongly activated, in a process that is accompanied by enrichment of activating histone marks (H3K4me3, H3ac, and H3K27ac) at the P1 promoter region. Histone mark analysis showed that SP7 gene promoter is robustly enriched in epigenetic repressive marks that may explain its poor transcriptional response to osteoblast differentiating media. Together, these results point to critical regulatory steps during epigenetic control of WJ‐MSCs osteogenic lineage commitment that are relevant for future applications in regenerative medicine. J. Cell. Physiol. 232: 2519–2527, 2017. © 2016 Wiley Periodicals, Inc. Wharton's Jelly Mesenchymal Stem Cells (WJ‐MSCs) are an attractive potential source of multipotent cells for tissue replacement therapies. Here, we study the ability of WJ‐MSCs to engage the osteoblast differentiation by analyzing epigenetic signatures at the promoter regions of the two master regulators of this process, RUNX2/P57 and SP7. We find that the RUNX2/p57, but not SP7, gene is expressed in WJ‐MSCs stimulated to differentiate to osteoblasts. This process is accompanied by the enrichment of activating histone marks at the RUNX2‐P57 P1 promoter and maintenance of epigenetic silencing histone marks at the SP7 promoter.

  March 28, 2017   doi: 10.1002/jcp.25627   open full text
• Super‐resolution structure of DNA significantly differs in buccal cells of controls and Alzheimer's patients.
  Angeles Garcia, David Huang, Amanda Righolt, Christiaan Righolt, Maria Carmela Kalaw, Shubha Mathur, Elizabeth McAvoy, James Anderson, Angela Luedke, Justine Itorralba, Sabine Mai.
  Journal of Cellular Physiology. March 28, 2017
  The advent of super‐resolution microscopy allowed for new insights into cellular and physiological processes of normal and diseased cells. In this study, we report for the first time on the super‐resolved DNA structure of buccal cells from patients with Alzheimer's disease (AD) versus age‐ and gender‐matched healthy, non‐caregiver controls. In this super‐resolution study cohort of 74 participants, buccal cells were collected and their spatial DNA organization in the nucleus examined by 3D Structured Illumination Microscopy (3D‐SIM). Quantitation of the super‐resolution DNA structure revealed that the nuclear super‐resolution DNA structure of individuals with AD significantly differs from that of their controls (p < 0.05) with an overall increase in the measured DNA‐free/poor spaces. This represents a significant increase in the interchromatin compartment. We also find that the DNA structure of AD significantly differs in mild, moderate, and severe disease with respect to the DNA‐containing and DNA‐free/poor spaces. We conclude that whole genome remodeling is a feature of buccal cells in AD. Figure 1 summarizes the super‐resolved DNA structure of buccal cells in Alzheimer's patients and their age‐ and gender‐matched controls.

  March 28, 2017   doi: 10.1002/jcp.25751   open full text
• MicroRNA‐1 overexpression blunts cardiomyocyte hypertrophy elicited by thyroid hormone.
  Gabriela Placoná Diniz, Caroline Antunes Lino, Camila Rodrigues Moreno, Nathalia Senger, Maria Luiza Morais Barreto‐Chaves.
  Journal of Cellular Physiology. March 28, 2017
  It is well‐known that increased thyroid hormone (TH) levels induce cardiomyocyte growth. MicroRNAs (miRNAs) have been identified as key players in cardiomyocyte hypertrophy, which is associated with increased risk of heart failure. In this study, we evaluated the miR‐1 expression in TH‐induced cardiac hypertrophy, as well as the potential involvement of miR‐1 in cardiomyocyte hypertrophy elicited by TH in vitro. The possible role of type 1 angiotensin II receptor (AT1R) in the effect promoted by TH in miR‐1 expression was also evaluated. Neonatal rat cardiac myocytes (NRCMs) were treated with T3 for 24 hr and Wistar rats were subjected to hyperthyroidism for 14 days combined or not with AT1R blocker. Real Time RT‐PCR analysis indicated that miR‐1 expression was decreased in cardiac hypertrophy in response to TH in vitro and in vivo, and this effect was unchanged by AT1R blocker. In addition, HDAC4, which is target of miR‐1, was increased in NRCMs after T3 treatment. A gain‐of‐function study revealed that overexpression of miR‐1 prevented T3‐induced cardiomyocyte hypertrophy and reduced HADC4 mRNA levels in NRCMs. In vivo experiments confirmed the downregulation of miR‐1 in cardiac tissue from hyperthyroid animals, which was accompanied by increased HDAC4 mRNA levels. In addition, HDAC inhibitor prevented T3‐induced cardiomyocyte hypertrophy. Our data reveal a new mechanistic insight into cardiomyocyte growth in response to TH, suggesting that miR‐1 plays a role in cardiomyocyte hypertrophy induced by TH potentially via targeting HADC4. A gain‐of‐function study revealed that overexpression of miR‐1 prevented T3‐induced cardiomyocyte hypertrophy and reduced HADC4 mRNA levels in neonatal cardiomyocytes. In vivo experiments confirmed the downregulation of miR‐1 in cardiac tissue from hyperthyroid animals, which was accompanied by increased HDAC4 mRNA levels. HDAC inhibition prevented T3‐induced cardiomyocyte hypertrophy revealing a new mechanistic insight into cardiomyocyte growth in response to TH.

  March 28, 2017   doi: 10.1002/jcp.25781   open full text
• Novel circulating biomarkers for non‐alcoholic fatty liver disease: A systematic review.
  Amirhossein Sahebkar, Elena Sancho, David Abelló, Jordi Camps, Jorge Joven.
  Journal of Cellular Physiology. March 28, 2017
  Currently, a liver biopsy remains the only reliable way to precisely diagnose non‐alcoholic fatty liver disease (NAFLD) and establish the severity of liver injury, presence of fibrosis, and architecture remodeling. However, the cost and the intrinsic invasive procedure of a liver biopsy rules it out as a gold standard diagnostic test, and the imaging test are not the best choice due to the price, and currently is being refined. The lack of a biomarker of NAFLD pushes to develop this new line of research. The aim of the present systematic review is to clarify and update all the NAFLD biomarkers described in the literature until recently. We highlight α‐ketoglutarate and CK18‐F as currently the best potential biomarker of NAFLD. However, due to methodological differences, we propose the implementation of international, multicenter, multiethnic studies with larger population size, and biopsy proven NAFLD diagnosis to analyze and compare α‐ketoglutarate and CK18‐F as potential biomarkers of the silent evolution of NAFLD. The lack of a biomarker of NAFLD pushes to develop this new line of research. The aim of the present systematic review is to clarify and update all the NAFLD biomarkers described in the literature until recently. We highlight α‐ketoglutarate and CK18‐F as currently the best potential biomarker of NAFLD.

  March 28, 2017   doi: 10.1002/jcp.25779   open full text
• Steady state peripheral blood provides cells with functional and metabolic characteristics of real hematopoietic stem cells.
  Antonin Bourdieu, Maryse Avalon, Véronique Lapostolle, Sadek Ismail, Margaux Mombled, Christelle Debeissat, Marianne Guérinet, Pascale Duchez, Jean Chevaleyre, Marija Vlaski‐Lafarge, Arnaud Villacreces, Vincent Praloran, Zoran Ivanovic, Philippe Brunet de la Grange.
  Journal of Cellular Physiology. March 28, 2017
  Hematopoietic stem cells (HSCs), which are located in the bone marrow, also circulate in cord and peripheral blood. Despite high availability, HSCs from steady state peripheral blood (SSPB) are little known and not used for research or cell therapy. We thus aimed to characterize and select HSCs from SSPB by a direct approach with a view to delineating their main functional and metabolic properties and the mechanisms responsible for their maintenance. We chose to work on Side Population (SP) cells which are highly enriched in HSCs in mouse, human bone marrow, and cord blood. However, no SP cells from SSBP have as yet been characterized. Here we showed that SP cells from SSPB exhibited a higher proliferative capacity and generated more clonogenic progenitors than non‐SP cells in vitro. Furthermore, xenotransplantation studies on immunodeficient mice demonstrated that SP cells are up to 45 times more enriched in cells with engraftment capacity than non‐SP cells. From a cell regulation point of view, we showed that SP activity depended on O2 concentrations close to those found in HSC niches, an effect which is dependent on both hypoxia‐induced factors HIF‐1α and HIF‐2α. Moreover SP cells displayed a reduced mitochondrial mass and, in particular, a lower mitochondrial activity compared to non‐SP cells, while they exhibited a similar level of glucose incorporation. These results provided evidence that SP cells from SSPB displayed properties of very primitive cells and HSC, thus rendering them an interesting model for research and cell therapy. We provided evidence that Side Population (SP) cells from Steady State Peripheral Blood (SSPB) displayed properties of very primitive cells and HSC based on in vitro and in vivo functional assays. Moreover these SP cells are better maintained during culture at low O2 concentrations, and they displayed a reduced mitochondrial mass and mitochondrial activity compared to non‐SP cells, while exhibiting a similar level of glucose incorporation. These cells are thus an interesting model for research and cell therapy.

  March 28, 2017   doi: 10.1002/jcp.25881   open full text
• Advanced glycation end products (AGE) potentiates cell death in p53 negative cells via upregulaion of NF‐kappa B and impairment of autophagy.
  Neeharika Verma, Sunil K. Manna.
  Journal of Cellular Physiology. March 27, 2017
  Accumulation of advanced glycation end products (AGE) in diabetic patients and ageing people due to excess availability of simple 3‐ or 4‐carbon sugars, is well‐known. AGE has multiple deleterious effects including age‐related disorders, apoptosis, inflammation, and obesity. We have found that AGE increases autophagy but the sustained amount of autophagosomes is observed till 3 days without maturation. It is important to understand the underlying mechanism of AGE‐mediated signaling responsible for impairment of autophagy and its correlation to the induction of several adverse effects. We have identified cross talk between autophagy and apoptosis upon AGE stimulation, specifically in p53 negative cells. AGE impairs autophagosomes' clearance in p53 negative cells as observed with an autophagosome maturation blocker‐bafilomycinA1 treated cells. This autophagy impairment is well supported by upregulation and overexpression of NF‐κB in these p53 negative cells. Autophagy impairment acts as a switch to initiate apoptosis via regulation of NF‐κB and its dependent genes. Increase in the expression of NF‐κB‐dependent NEDD4, an E3 ubiquitin ligase, which targets Beclin1 for cleavage is also evident. Beclin1 interacts with Bcl‐2, an anti‐apoptotic protein thereby engaging it to facilitate apoptosis upon AGE stimulation. For the first time, we are providing data that NF‐κB targeted cell signaling is involved in AGE‐mediated autophagy impairment in p53 negative/null cells. The p53 acts antagonistically to prevent this impairment. This study will help to control the AGE‐mediated detrimental effects associated with ageing and lysosomal storage disorders. NF‐κB targeted cell signaling is involved in AGE‐mediated autophagy impairment in p53 negative/null cells. The p53 acts antagonistically to prevent this impairment. This study will help to control the AGE‐mediated detrimental effects associated with ageing and lysosomal storage disorders.

  March 27, 2017   doi: 10.1002/jcp.25828   open full text
• Distinctive expression pattern of cystathionine‐β‐synthase and cystathionine‐γ‐lyase identifies mesenchymal stromal cells transition to mineralizing osteoblasts.
  Laura Gambari, Gina Lisignoli, Elena Gabusi, Cristina Manferdini, Francesca Paolella, Anna Piacentini, Francesco Grassi.
  Journal of Cellular Physiology. March 27, 2017
  Mesenchymal stromal cells (MSCs) are key players in the repair or regeneration of the damaged bone tissue. However, heterogeneity exists between MSCs derived from different donors in their bone formation ability both in vitro and in vivo. The identification of markers defining MSCs with different functional phenotypes is fundamental to maximize their clinical potential. In our previous in vivo study, impaired expression in MSCs of cystathionine‐β‐synthase (CBS) and cystathionine‐γ‐lyase (CSE), the two key enzymes in the catabolic pathway of homocysteine, was associated to decreased bone formation and to the onset of osteoporosis in mice. Here, we investigated whether osteogenic differentiation of human MSCs (hMSCs) modulates the expression of CBS and CSE. The expression of CBS and CSE was also assessed during chondrogenesis to confirm the specificity of their expression during osteogenesis. hMSCs displayed a heterogeneous mineralizing capacity between donors (70% of the samples mineralized, while 30% did not mineralize). Inducible expression of CBS and CSE was found to be associated with a mineralizing phenotype in hMSCs. In particular, up‐regulation of CSE was restricted to hMSCs undergoing mineralization. During chondrogenesis, CBS was significantly up‐regulated while CSE expression was not affected. Ex‐vivo findings confirmed that mature h‐osteoblasts (hOBs) show consistently higher expression of CBS and CSE than hMSCs. Our data provide the first evidence that the expression of CBS and CSE in hMSCs closely correlates with the transition of hMSCs toward the osteoblastic phenotype and that CSE may constitute a novel marker of osteogenic differentiation. Human mesenchymal stromal cells (hMSCs) play a pivotal role in the regeneration of bone tissue, but are affected by a marked heterogeneity in functional response to osteogenic stimuli. Inducible expression of CBS and CSE was found to be associated with a mineralizing phenotype in hMSCs and their expression correlates with the transition of hMSCs towards the osteoblastic phenotype. CSE may constitute a novel marker of osteogenic differentiation.

  March 27, 2017   doi: 10.1002/jcp.25825   open full text
• Roles of Notch1 Signaling in Regulating Satellite Cell Fates Choices and Postnatal Skeletal Myogenesis.
  Tizhong Shan, Ziye Xu, Weiche Wu, Jiaqi Liu, Yizhen Wang.
  Journal of Cellular Physiology. March 27, 2017
  Adult skeletal muscle stem cells, also called satellite cells, are indispensable for the growth, maintenance, and regeneration of the postnatal skeletal muscle. Satellite cells, predominantly quiescent in mature resting muscles, are activated after skeletal muscle injury or degeneration. Notch1 signaling is an evolutionarily conserved pathway that plays crucial roles in satellite cells homeostasis and postnatal skeletal myogenesis and regeneration. Activation of Notch1 signaling promotes the muscle satellite cells quiescence and proliferation, but inhibits differentiation of muscle satellite cells. Notably, the new roles of Notch1 signaling during late‐stage of skeletal myogenesis including in post‐differentiation myocytes and post‐fusion myotubes have been recently reported. Here, we mainly review and discuss the regulatory roles of Notch1 in regulating satellite cell fates choices and skeletal myogenesis. J. Cell. Physiol. 9999: 1–4, 2016. © 2016 Wiley Periodicals, Inc. Notch1 signaling is an evolutionarily conserved pathway that plays crucial roles in satellite cells homeostasis and postnatal skeletal myogenesis. Recently, the new roles of Notch1 signaling during late‐stage of skeletal myogenesis including in post‐differentiation myocytes and post‐fusion myotubes have been reported. Here, we mainly review and discuss the regulatory roles of Notch1 in regulating satellite cell fates choices and skeletal myogenesis.

  March 27, 2017   doi: 10.1002/jcp.25730   open full text
• The role of various peroxisome proliferator‐activated receptors and their ligands in clinical practice.
  Giuseppe Derosa, Amirhossein Sahebkar, Pamela Maffioli.
  Journal of Cellular Physiology. March 27, 2017
  Peroxisome proliferator‐activated receptors (PPARs) are ligand‐activated transcription factors involved in several physiological processes including modulation of cellular differentiation, development, metabolism of carbohydrates, lipids, proteins, and tumorigenesis. The aim of this review is to examine how different PPAR ligands act, and discuss their use in clinical practice. PPAR ligands have a lot of effects and applications in clinical practice. Some PPAR ligands such as fibrates (PPAR‐α ligands) are currently used for the treatment of dyslipidemia, while pioglitazone and rosiglitazone (PPAR‐γ ligands) are anti‐diabetic and insulin‐sensitizing agents. Regarding new generation drugs, acting on both α/γ, β/δ, or α/δ receptors simultaneously, preliminary data on PPAR‐α/γ dual agonists revealed a positive effect on lipid profile, blood pressure, atherosclerosis, inflammation, and anti‐coagulant effects, while the overexpression of PPAR‐β/δ seems to prevent obesity and to decrease lipid storage in cardiac cells. Finally, PPAR‐α/δ dual agonist induces resolution of nonalcoholic steatohepatitis without fibrosis worsening. Peroxisome proliferator‐activated receptors (PPARs) are ligand‐activated transcription factors involved in several physiological processes. PPAR ligands have a lot of effects and applications in clinical practice. The aim of this review is to examine how different PPAR ligands act, and discuss their use in clinical practice.

  March 27, 2017   doi: 10.1002/jcp.25804   open full text
• NOR1 Suppresses Cancer Stem‐Like Cells Properties of Tumor Cells via the Inhibition of the AKT‐GSK‐3β‐Wnt/β‐catenin‐ALDH1A1 Signal Circuit.
  Wei Wang, Mei Yi, Shengnan Chen, Junjun Li, Haijing Zhang, Wei Xiong, Guiyuan Li, Xiaoling Li, Bo Xiang.
  Journal of Cellular Physiology. March 27, 2017
  Cancer stem cells (CSCs) play a key role in tumor radiotherapy and chemotherapy resistance, relapse, and metastasis, and are primarily maintained in a resting state in vivo. The failure of conventional therapies to target CSCs is the main cause of treatment failure. The discovery of CSCs in nasopharyngeal carcinoma (NPC) tumors is becoming more prevalent; however, the understanding of the mechanisms underlying the maintenance of tumor stemness is still limited. We previously cloned NOR1, a tumor suppressor gene downregulated in NPC cell lines and tissues. In this study, we demonstrate that Wnt/β‐catenin and ALDH1A1 form a signal circuit and that NOR1 antagonizes the tumor stem cell‐like phenotype in NPC cell lines: the ectopic overexpression of NOR1 reduced β‐catenin and ALDH1A1 expression; β‐catenin/TCF4 targeted the regulation of ALDH1A1 transcription in NPC cells; silencing ALDH1A1 reduced AKT (total and phosphorylated) and GSK‐3β (phosphorylated) expression; and eventually feedback decreased β‐catenin expression levels. We also found that NOR1 expression decreased cancer stem‐like cell properties of NPC cells, reduced their ability to form tumor spheroids in vitro, reduced tumorigenicity in nude mice in vivo, and increased sensitivity to chemotherapy agents. Taken together, our findings illustrated a new function of NOR1 that suppresses cancer stem‐like cell properties in tumor cells by inhibiting the AKT‐GSK‐3β‐Wnt/β‐catenin‐ALDH1A1 signal circuit. The study suggests that NOR1 deletion expression in NPC cells may be a potential molecular target for cancer stem cell therapy. J. Cell. Physiol. 232: 2829–2840, 2017. © 2016 Wiley Periodicals, Inc. Cancer stem cells (CSCs) play a key role in tumor radiotherapy and chemotherapy resistance, relapse, and metastasis. Our findings illustrated a new function of NOR1 that suppresses cancer stem‐like cell properties in tumor cells by inhibiting the AKT‐GSK‐3β‐Wnt/β‐catenin‐ALDH1A1 signal circuit.

  March 27, 2017   doi: 10.1002/jcp.25706   open full text
• Methylated of genes behaving as potential biomarkers in evaluating malignant degree of glioblastoma.
  Wan‐Shun Wen, Sheng‐Li Hu, Zhibing Ai, Lin Mou, Jing‐Min Lu, Sen Li.
  Journal of Cellular Physiology. March 27, 2017
  Abnormal methylation genes usually act as oncogenes or anti‐oncogenes in the occurrence and development of tumor, indicating their potential role as biomarkers in the evaluation of malignant tumor. However, the research on methylation's association with glioblastoma was rare. We attempted to figure out whether the methylation of genes could serve as the biomarker in evaluating the malignant degree of GBM. Methylation microarray data of 275 GBM patients have been downloaded from The Cancer Genome Atlas (TCGA) dataset. Logistic regression was used to find the methylated genes associated with the malignant degree of patients with the tumor. Functional enrichment analysis and network analysis were further performed on these selected genes. A total of 668, 412, 470, and 620 genes relevant with the methylation or demethylation were found to be associated with the malignant degree, Grades 1–4 of tumor. The higher the degree of malignant tumor, the higher of its methylation degree of its corresponding genes. GO and KEGG analysis results showed that these methylated genes were enriched in many functions as cell adhesion, abnormal transcription, and cell cycle disorder, etc. Of note, CCL11 and LCN11 were found to be significantly related to the progression of GBM. Critical genes associated with cell cycle as CCL11 and LCN1 may play essential roles in the occurrence, development, and transition of glioblastoma. More research was needed to explore its potential molecular mechanism. Abnormal methylation genes usually act as oncogenes or anti‐oncogenes in the occurrence and development of tumor, indicating their potential role as biomarkers in the evaluation of malignant tumor.A total of 668, 412, 470 and 620 genes relevant with the methylation or demethylation were found to be associated with the malignant degree, Grades 1–4 of glioblastoma. Of note, CCL11 and LCN11 were found to be significantly related to the progression of GBM.

  March 27, 2017   doi: 10.1002/jcp.25831   open full text
• Mouse Oocytes Acquire Mechanisms That Permit Independent Cell Volume Regulation at the End of Oogenesis.
  Samantha Richard, Alina P. Tartia, Detlev Boison, Jay M. Baltz.
  Journal of Cellular Physiology. March 27, 2017
  Mouse embryos employ a unique mechanism of cell volume regulation in which glycine is imported via the GLYT1 transporter to regulate intracellular osmotic pressure. Independent cell volume regulation normally becomes active in the oocyte after ovulation is triggered. This involves two steps: the first is the release of the strong adhesion between the oocyte and zona pellucida (ZP) while the second is the activation of GLYT1. In fully‐grown oocytes, release of adhesion and GLYT1 activation also occur spontaneously in oocytes removed from the follicle. It is unknown, however, whether the capacity to release oocyte‐ZP adhesion or activate GLYT1 first arises in the oocyte after ovulation is triggered or instead growing oocytes already possess these capabilities but they are suppressed in the follicle. Here, we assessed when during oogenesis oocyte‐ZP adhesion can be released and when GLYT1 can be activated, with adhesion assessed by an osmotic assay and GLYT1 activity determined by [3H]‐glycine uptake. Oocyte‐ZP adhesion could not be released by growing oocytes until they were nearly fully grown. Similarly, the amount of GLYT1 activity that can be elicited in oocytes increased sharply at the end of oogenesis. The SLC6A9 protein that is responsible for GLYT1 activity and Slc6a9 transcripts are present in growing oocytes and increased over the course of oogenesis. Furthermore, SLC6A9 becomes localized to the oocyte plasma membrane as the oocyte grows. Thus, oocytes acquire the ability to regulate their cell volume by releasing adhesion to the ZP and activating GLYT1 as they approach the end of oogenesis. J. Cell. Physiol. 232: 2436–2446, 2017. © 2016 Wiley Periodicals, Inc. Independent cell volume regulation in the mouse oocyte normally becomes active after ovulation is triggered in a two‐step process: the strong adhesion of the oocyte to the surrounding zona pellucida is released and glycine transport via the GLYT1 transporter is activated. Both mechanisms develop at the end of oocyte growth but remain suppressed within the ovarian follicle.

  March 27, 2017   doi: 10.1002/jcp.25581   open full text
• ­NOD Mice Having a Lyn Tyrosine Kinase Mutation Exhibit Abnormal Neutrophil Chemotaxis.
  Yue Wu, Michael Hannigan, Lijun Zhan, Joseph A. Madri, Chi‐Kuang Huang.
  Journal of Cellular Physiology. March 27, 2017
  Neutrophils from NOD (Non‐Obese Diabetic) mice exhibited reduced migration speed, decreased frequency of directional changes, and loss of directionality during chemotaxis (compared to wild‐type [WT] C57BL/6 mice). Additionally, F‐actin of chemotaxing NOD neutrophils failed to orient toward the chemoattractant gradient and NOD neutrophil adhesion was impaired. A point mutation near the autophosphorylation site of Lyn in NOD mice was identified. Point mutations of G to A (G1412 in LynA and G1199 in LynB) cause a change of amino acid E393 (glutamic acid) to K (lysine) in LynA (E393→K) (E372 of LynB), affecting fMLP‐induced tyrosine phosphorylation. These data indicate that the Lyn mutation in NOD neutrophils is likely responsible for dysregulation of neutrophil adhesion and directed migration, implying the role of Lyn in modulating diabetic patient's susceptibility to bacterial and fungal infections. J. Cell. Physiol. 232: 1689–1695, 2017. © 2016 Wiley Periodicals, Inc. A specific Lyn kinase mutation has been documented in NOD mice which affects neutrophil directed migration. This may thus affect response to infections in this murine model of diabetes.

  March 27, 2017   doi: 10.1002/jcp.25583   open full text
• Vertebrate Lonesome Kinase Regulated Extracellular Matrix Protein Phosphorylation, Cell Shape, and Adhesion in Trabecular Meshwork Cells.
  Rupalatha Maddala, Nikolai P. Skiba, Ponugoti Vasantha Rao.
  Journal of Cellular Physiology. March 27, 2017
  Glaucoma, a leading cause of irreversible blindness, is commonly associated with elevated intraocular pressure (IOP) due to impaired aqueous humor (AH) drainage through the trabecular meshwork (TM). Although dysregulated production and organization of extracellular matrix (ECM) is presumed to increase resistance to AH outflow and elevate IOP by altering TM cell contractile and adhesive properties, it is not known whether regulation of ECM protein phosphorylation via the secretory vertebrate lonesome kinase (VLK) influences TM cellular characteristics. Here, we tested this possibility. Experiments carried out in this study reveal that the 32 kDa protein is a prominent VLK isoform detectable in lysates and conditioned media (CM) of human TM cells. Increased levels of VLK were observed in CM of TM cells subjected to cyclic mechanical stretch, or treated with dexamethasone, TGF‐β2, and TM cells expressing constitutively active RhoA GTPase. Downregulation of VLK expression in TM cells using siRNA decreased tyrosine phosphorylation (TyrP) of ECM proteins and focal adhesions, and induced changes in cell shape in association with reduced levels of actin stress fibers and phospho‐paxillin. VLK was also demonstrated to regulate TGF‐β2‐induced TyrP of ECM proteins. Taken together, these results suggest that VLK secretion can be regulated by external cues, intracellular signal proteins, and mechanical stretch, and VLK can in turn regulate TyrP of ECM proteins secreted by TM cells and control cell shape, actin stress fibers, and focal adhesions. These observations indicate a potential role for VLK in homeostasis of AH outflow and IOP, and in the pathobiology of glaucoma. J. Cell. Physiol. 232: 2447–2460, 2017. © 2016 Wiley Periodicals, Inc. Our results suggest that VLK secretion can be regulated by external cues, intracellular signal proteins, and mechanical stretch, and VLK can in turn regulate TyrP of ECM proteins secreted by TM cells and control cell shape, actin stress fibers, and focal adhesions. These observations indicate a potential role for VLK in homeostasis of AH outflow and IOP, and in the pathobiology of glaucoma.

  March 27, 2017   doi: 10.1002/jcp.25582   open full text
• Mechanisms involved in enhancement of matrix metalloproteinase‐9 expression in macrophages by interleukin‐33.
  Wataru Ariyoshi, Toshinori Okinaga, Wichida Chaweewannakorn, Sumio Akifusa, Tatsuji Nisihara.
  Journal of Cellular Physiology. March 27, 2017
  Endothelial transmigration of macrophages is accomplished by matrix metalloproteinase (MMP)‐induced degradation of the basement membrane and extracellular matrix components. Macrophages upregulate MMP‐9 expression and secretion upon immunological challenges and require its activity for migration during inflammatory responses. Interleukin (IL)‐33 is a recently discovered pro‐inflammatory cytokine that belongs to the IL‐1 family. The aim of this study was to elucidate the mechanisms underlying IL‐33‐induced MMP‐9 expression in the mouse monocyte/macrophage line RAW264.7. IL‐33 increased MMP‐9 mRNA and protein expression in RAW264.7 cells. Blockage of IL‐33‐IL‐33 receptor (ST2L) binding suppressed IL‐33‐mediated induction of MMP‐9. IL‐33 induced phosphorylation and nuclear translocation of extracellular signal‐regulated kinase 1/2 (ERK1/2) and nuclear factor‐kappa B (NF‐κB). Chromatin immunoprecipitation indicated that IL‐33 increased c‐fos recruitment to the MMP‐9 promoter. Reporter assay findings also revealed that IL‐33 stimulated the transcriptional activity of activator protein 1 (AP‐1). Pre‐treatment of the cells with a specific inhibitor of ERK1/2 and NF‐κB attenuated the IL‐33‐induced activation of AP‐1 subunits, transcriptional activity of AP‐1, and expression of MMP‐9. We also demonstrated that ERK‐dependent activation of cAMP response element binding protein (CREB) is a key step for AP‐1 activation by IL‐33. These results indicate an essential role of ERK/CREB and NF‐κB cascades in the induction of MMP‐9 in monocytes/macrophages through AP‐1 activation. IL‐33‐ST2L interaction increased MMP‐9 expression in RAW264.7 cells. ERK/CREB and NF‐κB cascades have an essential role in the induction of MMP‐9 in monocytes/macrophages through AP‐1 activation.

  March 27, 2017   doi: 10.1002/jcp.25809   open full text
• Meta‐Analysis of Transcriptome Regulation During Induction to Cardiac Myocyte Fate From Mouse and Human Fibroblasts.
  Shima Rastegar‐Pouyani, Niusha Khazaei, Ping Wee, Moein Yaqubi, Abdulshakour Mohammadnia.
  Journal of Cellular Physiology. March 24, 2017
  Ectopic expression of a defined set of transcription factors (TFs) can directly convert fibroblasts into a cardiac myocyte cell fate. Beside inefficiency in generating induced cardiomyocytes (iCMs), the molecular mechanisms that regulate this process remained to be well defined. The main purpose of this study was to provide better insight on the transcriptome regulation and to introduce a new strategy for candidating TFs for the transdifferentiation process. Eight mouse and three human high quality microarray data sets were analyzed to find differentially expressed genes (DEGs), which we integrated with TF‐binding sites and protein–protein interactions to construct gene regulatory and protein–protein interaction networks. Topological and biological analyses of constructed gene networks revealed the main regulators and most affected biological processes. The DEGs could be categorized into two distinct groups, first, up‐regulated genes that are mainly involved in cardiac‐specific processes and second, down‐regulated genes that are mainly involved in fibroblast‐specific functions. Gata4, Mef2a, Tbx5, Tead4 TFs were identified as main regulators of cardiac‐specific gene expression program; and Trp53, E2f1, Myc, Sfpi1, Lmo2, and Meis1 were identified as TFs which mainly regulate the expression of fibroblast‐specific genes. Furthermore, we compared gene expression profiles and identified TFs between mouse and human to find the similarities and differences. In summary, our strategy of meta‐analyzing the data of high‐throughput techniques by computational approaches, besides revealing the mechanisms involved in the regulation of the gene expression program, also suggests a new approach for increasing the efficiency of the direct reprogramming of fibroblasts into iCMs. J. Cell. Physiol. 232: 2053–2062, 2017. © 2016 Wiley Periodicals, Inc. The DEGs could be categorized into two distinct groups, first, up‐regulated genes that are mainly involved in cardiac‐specific processes and second, down‐regulated genes that are mainly involved in fibroblast‐specific functions. Gata4, Mef2a, Tbx5, Tead4 TFs were identified as main regulators of cardiac‐specific gene expression program; and Trp53, E2f1, Myc, Sfpi1, Lmo2, and Meis1 were identified as TFs which mainly regulate the expression of fibroblast‐specific genes. Furthermore, we compared gene expression profiles and identified TFs between mouse and human to find the similarities and differences.

  March 24, 2017   doi: 10.1002/jcp.25580   open full text
• Molecular Imaging of Huntington's Disease.
  Andrea Ciarmiello, Giampiero Giovacchini, Elisabetta Giovannini, Patrizia Lazzeri, Elisa Borsò, Antonio Mannironi, Luigi Mansi.
  Journal of Cellular Physiology. March 24, 2017
  The onset and the clinical progression of Huntington Disease (HD) is influenced by several events prompted by a genetic mutation that affects several organs tissues including different regions of the brain. In the last decades years, Positron Emission Tomography (PET) and Magnetic Resonance Imaging (MRI) helped to deepen the knowledge of neurodegenerative mechanisms that guide to clinical symptoms. Brain imaging with PET represents a tool to investigate the physiopathology occurring in the brain and it has been used to predict the age of onset of the disease and to evaluate the therapeutic efficacy of new drugs. This article reviews the contribution of PET and MRI in the research field on Huntington's disease, focusing in particular on some most relevant achievements that have helped recognize the molecular changes, the clinical symptoms and evolution of the disease. J. Cell. Physiol. 232: 1988–1993, 2017. © 2016 Wiley Periodicals, Inc. The onset and the clinical progression of Huntington Disease (HD) is influenced by several events prompted by a genetic mutation that affects several organs tissues including different regions of the brain. In the last decades years, Positron Emission Tomography (PET) and Magnetic Resonance Imaging (MRI) helped to deepen the knowledge of neurodegenerative mechanisms that guide to clinical symptoms. Brain imaging with PET represents a tool to investigate the physiopathology occurring in the brain and it has been used to predict the age of onset of the disease and to evaluate the therapeutic efficacy of new drugs. This article reviews the contribution of PET and MRI in the research field on Huntington's disease, focusing in particular on some most relevant achievements that have helped recognize the molecular changes, the clinical symptoms, and evolution of the disease.

  March 24, 2017   doi: 10.1002/jcp.25666   open full text
• Interactive Effect of Corticosterone and Lactate on Regulation of Testosterone Production in Rat Leydig Cells.
  Chih‐Chieh Chen, Chien‐Wei Chen, Po‐Han Lin, Jou‐Chun Chou, Ting‐Chun Weng, Cai‐Yun Jian, Sindy Hu, Wei‐Ho Lai, Fu‐Kong Lieu, Shyi‐Wu Wang, Paulus S. Wang.
  Journal of Cellular Physiology. March 24, 2017
  The increasing intensity of exercise enhanced corticosterone and lactate production in both humans and rodents. Our previous studies also demonstrated that lactate could stimulate testosterone production in vivo and in vitro. However, the production of testosterone in response to combined corticosterone and lactate on Leydig cells, and underlying molecular mechanisms are remained unclear. This study investigated the changes in testosterone levels of Leydig cells upon exposure to lactate, corticosterone or combination of both, and revealed the detailed mechanisms. Leydig cells were isolated from rat testes, and treated with different concentrations of lactate (2.5–20 mM), cortiosterone (10−9–10−4 M) and lactate plus corticosterone. The production of testosterone were assayed by radioimmunoassay, and the key molecular proteins, including luteinizing hormone receptor (LHR), protein kinase A (PKA), steroidogenic acute regulatory protein (StAR), and cholesterol P450 side‐chain cleavage enzyme (P450scc) involved in testosterone production were performed by Western blot. Results showed that testosterone levels were significantly increased with lactate, while decresed with corticosterone and lactate plus corticosterone treatment. Protein expressions of LHR and P450scc were upregulated with lactate treatment. However, PKA and P450scc were downregulated by lactate plus corticosterone treatment. This downregulation was followed by decreased testoterone levels in Leydig cells. Furthermore, acetylated cAMP, which activates testosterone production was increased with lactate, but not altered by conrtiosterone. Our findings conclude that corticosterone may interfere with lactate, and restrict lactate‐stimulated testosterone production in Leydig cells. J. Cell. Physiol. 232: 2135–2144, 2017. © 2016 Wiley Periodicals, Inc. Lactate activates the LHR‐cAMP‐P450scc pathway to increase endogenous testosterone production. Treatment of CORT down‐regulates LHR‐cAMP‐PKA‐P450scc pathway to diminish testosterone production. Activation of intracellular adenylyl cyclase, PKA and P450scc pathways are critical to increase endogenous testosterone production in Leydig cells.

  March 24, 2017   doi: 10.1002/jcp.25700   open full text
• Palmitoleic Acid Improves Metabolic Functions in Fatty Liver by PPARα‐Dependent AMPK Activation.
  Camila O. de Souza, Alexandre A.S. Teixeira, Luana A. Biondo, Edson A. Lima Junior, Helena A.P. Batatinha, Jose C. Rosa Neto.
  Journal of Cellular Physiology. March 24, 2017
  Background: Palmitoleic acid, since described as lipokine, increases glucose uptake by modulation of 5′AMP‐activated protein kinase (AMPK), as well as increasing lipolysis by activation of peroxisome proliferator‐activated receptor‐α (PPARα), in adipose tissue. However, in liver, the effects of palmitoleic acid on glucose metabolism and the role of PPARα remain unknown. Objective: To investigate whether palmitoleic acid improved the hepatic insulin sensitivity of obese mice. Methods: C57BL6 and PPARα knockout (KO) mice were fed for 12 weeks with a standard diet (SD) or high‐fat diet (HF), and in the last 2 weeks were treated with oleic or palmitoleic acid. Results: Palmitoleic acid promoted a faster uptake of glucose in the body, associated with higher insulin concentration; however, even when stimulated with insulin, palmitoleic acid did not modulate the insulin pathway (AKT, IRS). Palmitoleic acid increased the phosphorylation of AMPK, upregulated glucokinase and downregulated SREBP‐1. Regarding AMPK downstream, palmitoleic acid increased the production of FGF‐21 and stimulated the expression of PPARα. Palmitoleic acid treatment did not increase AMPK phosphorylation, modulate glucokinase or increase FGF‐21 in liver of PPARα KO mice. Conclusions: In mice fed with a high‐fat diet, palmitoleic acid supplementation stimulated the uptake of glucose in liver through activation of AMPK and FGF‐21, dependent on PPARα. J. Cell. Physiol. 232: 2168–2177, 2017. © 2016 Wiley Periodicals, Inc. Palmitoleic acid (PM) increased glucose uptake and modulated glucokinase in liver; Those effects are linked to higher PPARα expression, activation of AMPK and FGF‐21. Similar beneficial effects were not observed in liver of PPARα KO mice. Thus, activation of AMPK and FGF‐21 by PM in liver, are dependent of PPARα.

  March 24, 2017   doi: 10.1002/jcp.25715   open full text
• Opposite Effects of Mechanical Action of Fluid Flow on Proangiogenic Factor Secretion From Human Adipose‐Derived Stem Cells With and Without Oxidative Stress.
  Beatriz Bravo, Cira García de Durango, Álvaro González, Arancha R. Gortázar, Xavier Santos, Jerónimo Forteza‐Vila, Fernando Vidal‐Vanaclocha.
  Journal of Cellular Physiology. March 24, 2017
  Mechanical forces, hypoxia, and oxidative stress contribute to skin renewal, perfusion, and wound healing, but how are they regulating subcutaneous adipose‐derived stem cells (ASCs) in the inflammatory microenvironment associated to skin repair and disorders is unknown. In this study, ASCs were isolated from lipoaspirate samples from plastic surgery patients, primary cultured and their differentiation and secretion of a panel of cytokines with pronounced effects on skin repair and angiogenesis were studied under mechanical stimulation by intermittent fluid flow, 1% hypoxia and oxidative stress by glutathione (GSH) depletion with buthionine sulfoximine (BSO) treatment. Mechanical action of fluid flow did not alter mesenchymal phenotype of CD90+/CD29+/CD44+/CD34−/CD106−/CD45−ASCs; however, it remarkably induced ASC secretion of human umbilical vein endothelial cell (HUVEC) migration‐stimulating factors. Multiplex Luminex assay further confirmed an increased secretion of VEGF, G‐CSF, HGF, Leptin, IL‐8, PDGF‐BB, Angiopoietin‐2, and Follistatin from mechanically‐stimulated ASCs via cyclooxygenase‐2. Consistent with this mechanism, GSH depletion and hypoxia also increased ASC secretion of VEGF, IL‐8, leptin, Angiopoitein‐2, and PDGF‐BB. However, mechanical action of fluid flow abrogated VEGF and HUVEC migration‐stimulating activity from GSH‐depleted and hypoxic ASCs. Conversely, GSH depletion and hypoxia abrogated VEGF and HUVEC migration‐stimulating activity from mechano‐stimulated ASCs. Although mechanical action of fluid flow, hypoxia, and GSH‐depletion had independent proangiogenic‐stimulating activity on ASCs, mechanical stimulation had opposite effects on proangiogenic factor secretion from ASCs with and without oxidative stress. These data uncover the role of hypoxia and endogenous redox balance during the proangiogenic response of ASCs and other mesenchymal‐derived cell types to mechanical action of interstitial fluid flow. J. Cell. Physiol. 232: 2158–2167, 2017. © 2016 Wiley Periodicals, Inc. In this original work, we described the critical role of endogenous redox balance during the proangiogenic response of ASCs to mechanical forces. These data uncover the role of hypoxia and endogenous redox balance during the proangiogenic response of ASCs and other mesenchymal‐derived cell types to mechanical action of interstitial fluid flow.

  March 24, 2017   doi: 10.1002/jcp.25712   open full text
• Aging Selectively Modulates Vitamin C Transporter Expression Patterns in the Kidney.
  Katherine Forman, Fernando Martínez, Manuel Cifuentes, Romina Bertinat, Katterine Salazar, Francisco Nualart.
  Journal of Cellular Physiology. March 24, 2017
  In the kidney, vitamin C is reabsorbed from the glomerular ultrafiltrate by sodium‐vitamin C cotransporter isoform 1 (SVCT1) located in the brush border membrane of the proximal tubules. Although we know that vitamin C levels decrease with age, the adaptive physiological mechanisms used by the kidney for vitamin C reabsorption during aging remain unknown. In this study, we used an animal model of accelerated senescence (SAMP8 mice) to define the morphological alterations and aging‐induced changes in the expression of vitamin C transporters in renal tissue. Aging induced significant morphological changes, such as periglomerular lymphocytic infiltrate and glomerular congestion, in the kidneys of SAMP8 mice, although no increase in collagen deposits was observed using 2‐photon microscopy analysis and second harmonic generation. The most characteristic histological alteration was the dilation of intracellular spaces in the basolateral region of proximal tubule epithelial cells. Furthermore, a combination of laser microdissection, qRT‐PCR, and immunohistochemical analyses allowed us to determine that SVCT1 expression specifically increased in the proximal tubules from the outer strip of the outer medulla (segment S3) and cortex (segment S2) during aging and that these tubules also express GLUT1. We conclude that aging modulates vitamin C transporter expression and that renal over‐expression of SVCT1 enhances vitamin C reabsorption in aged animals that may synthesize less vitamin C. J. Cell. Physiol. 232: 2418–2426, 2017. © 2016 Wiley Periodicals, Inc. Although we know that vitamin C levels decrease with age, the adaptive physiological mechanisms used by the kidney for vitamin C reabsorption during aging remain unknown. In this study, we used an animal model of accelerated senescence (SAMP8 mice) to define the morphological alterations and aging‐induced changes in the expression of vitamin C transporters in renal tissue.

  March 24, 2017   doi: 10.1002/jcp.25504   open full text
• Lipocalin‐2 (NGAL) Attenuates Autophagy to Exacerbate Cardiac Apoptosis Induced by Myocardial Ischemia.
  Hye Kyoung Sung, Yee Kwan Chan, Meng Han, James Won Suk Jahng, Erfei Song, Eric Danielson, Thorsten Berger, Tak W. Mak, Gary Sweeney.
  Journal of Cellular Physiology. March 24, 2017
  Lipocalin‐2 (Lcn2; also termed neutrophil gelatinase‐associated lipocalin (NGAL)) levels correlate positively with heart failure (HF) yet mechanisms via which Lcn2 contributes to the pathogenesis of HF remain unclear. In this study, we used coronary artery ligation surgery to induce ischemia in wild‐type (wt) mice and this induced a significant increase in myocardial Lcn2. We then compared wt and Lcn2 knockout (KO) mice and observed that wt mice showed greater ischemia‐induced caspase‐3 activation and DNA damage measured by TUNEL than Lcn2KO mice. Analysis of autophagy by LC3 and p62 Western blotting, LC3 immunohistochemistry and transmission electron microscopy (TEM) indicated that Lcn2 KO mice had a greater ischemia‐induced increase in autophagy. Lcn2KO were protected against ischemia‐induced cardiac functional abnormalities measured by echocardiography. Upon treating a cardiomyocyte cell line (h9c2) with Lcn2 and examining AMPK and ULK1 phosphorylation, LC3 and p62 by Western blot as well as tandem fluorescent RFP/GFP‐LC3 puncta by immunofluorescence, MagicRed assay for lysosomal cathepsin activity and TEM we demonstrated that Lcn2 suppressed autophagic flux. Lcn2 also exacerbated hypoxia‐induced cytochromc c release from mitochondria and caspase‐3 activation. We generated an autophagy‐deficient H9c2 cell model by overexpressing dominant‐negative Atg5 and found significantly increased apoptosis after Lcn2 treatment. In summary, our data indicate that Lcn2 can suppress the beneficial cardiac autophagic response to ischemia and that this contributes to enhanced ischemia‐induced cell death and cardiac dysfunction. J. Cell. Physiol. 232: 2125–2134, 2017. © 2016 Wiley Periodicals, Inc. This study examined the regulation of autophagy by Lcn2 and the functional significance in the heart. Our data led us to conclude that, at least in part, via inhibition of cardiomyocyte autophagy Lcn2 exacerbated stress‐induced cell death and cardiac dysfunction.

  March 24, 2017   doi: 10.1002/jcp.25672   open full text
• MAPK3/1 participates in the activation of primordial follicles through mTORC1‐KITL signaling.
  Yu Zhao, Yu Zhang, Jia Li, Nana Zheng, Xiaoting Xu, Jing Yang, Guoliang Xia, Meijia Zhang.
  Journal of Cellular Physiology. March 24, 2017
  The majority of ovarian primordial follicles are preserved in a dormant state to maintain the female reproductive lifespan, and only a few primordial follicles are activated to enter the growing follicle pool in each wave. Recent studies have shown that primordial follicular activation depends on mammalian target of rapamycin complex 1 (mTORC1)‐KIT ligand (KITL) signaling in pre‐granulosa cells and its receptor (KIT)‐phosphoinositol 3 kinase (PI3K) signaling in oocytes. However, the upstream regulator of mTORC1 signaling is unclear. The results of the present study showed that the phosphorylated mitogen‐activated protein kinase3/1 (MAPK3/1) protein is expressed in some primordial follicles and all growing follicles. Culture of 3 days post‐parturition (dpp) ovaries with the MAPK3/1 signaling inhibitor U0126 significantly reduced the number of activated follicles and was accompanied by dramatically reduced granulosa cell proliferation and increased oocyte apoptosis. Western blot and immunofluorescence analyses showed that U0126 significantly decreased the phosphorylation levels of Tsc2, S6K1, and rpS6 and the expression of KITL, indicating that U0126 inhibits mTORC1‐KITL signaling. Furthermore, U0126 decreased the phosphorylation levels of Akt, resulting in a decreased number of oocytes with Foxo3 nuclear export. To further investigate MAPK3/1 signaling in primordial follicle activation, we used phosphatase and tensin homolog deleted on chromosome 10 (PTEN) inhibitor bpV(HOpic) to promote primordial follicle activation. In this model, U0126 also inhibited the activation of primordial follicles and mTORC1 signaling. Thus, these results suggest that MAPK3/1 participates in primordial follicle activation through mTORC1‐KITL signaling. Primordial follicular activation depends on mammalian target of rapamycin complex 1 (mTORC1)‐KIT ligand (KITL) signaling in pre‐granulosa cells and its receptor (KIT)‐phosphoinositol 3 kinase (PI3K) signaling in oocytes. Here, we have shown that MAPK3/1 signaling participates in the activation of primordial follicles through mTORC1 signaling and the expression of KITL in pre‐granulosa cells to activate the PI3K‐Akt pathway.

  March 24, 2017   doi: 10.1002/jcp.25868   open full text
• p62 Promotes Amino Acid Sensitivity of mTOR Pathway and Hepatic Differentiation in Adult Liver Stem/Progenitor Cells.
  Masakazu Sugiyama, Tomoharu Yoshizumi, Yoshihiro Yoshida, Yuki Bekki, Yoshihiro Matsumoto, Shohei Yoshiya, Takeo Toshima, Toru Ikegami, Shinji Itoh, Norifumi Harimoto, Shinji Okano, Yuji Soejima, Ken Shirabe, Yoshihiko Maehara.
  Journal of Cellular Physiology. March 24, 2017
  Autophagy is a homeostatic process regulating turnover of impaired proteins and organelles, and p62 (sequestosome‐1, SQSTM1) functions as the autophagic receptor in this process. p62 also functions as a hub for intracellular signaling such as that in the mammalian target of rapamycin (mTOR) pathway. Liver stem/progenitor cells have the potential to differentiate to form hepatocytes or cholangiocytes. In this study, we examined effects of autophagy, p62, and associated signaling on hepatic differentiation. Adult stem/progenitor cells were isolated from the liver of mice with chemically induced liver injury. Effects of autophagy, p62, and related signaling pathways on hepatic differentiation were investigated by silencing the genes for autophagy protein 5 (ATG5) and/or SQSTM1/p62 using small interfering RNAs. Hepatic differentiation was assessed based on increased albumin and hepatocyte nuclear factor 4α, as hepatocyte markers, and decreased cytokeratin 19 and SOX9, as stem/progenitor cell markers. These markers were measured using quantitative RT‐PCR, immunofluorescence, and Western blotting. ATG5 silencing decreased active LC3 and increased p62, indicating inhibition of autophagy. Inhibition of autophagy promoted hepatic differentiation in the stem/progenitor cells. Conversely, SQSTM1/p62 silencing impaired hepatic differentiation. A suggested mechanism for p62‐dependent hepatic differentiation in our study was activation of the mTOR pathway by amino acids. Amino acid activation of mTOR signaling was enhanced by ATG5 silencing and suppressed by SQSTM1/p62 silencing. Our findings indicated that promoting amino acid sensitivity of the mTOR pathway is dependent on p62 accumulated by inhibition of autophagy and that this process plays an important role in the hepatic differentiation of stem/progenitor cells. J. Cell. Physiol. 232: 2112–2124, 2017. © 2016 Wiley Periodicals, Inc. Inhibition of autophagy by siRNA for ATG5 promoted hepatic differentiation in liver stem/progenitor cells. Autophagic adaptor protein, p62, was an important molecule to promote hepatic differentiation by inhibition of autophagy. p62 promoted amino acid sensitivity of the mTOR pathway and hepatic differentiation in liver stem/progenitor cells.

  March 24, 2017   doi: 10.1002/jcp.25653   open full text
• T11TS repress gliomagenic apoptosis of bone marrow hematopoietic stem cells.
  Somnath Mondal, Iman Hazra, Ankur Datta, Omar Faruk Sk Md, Saibal Moitra, Santanu Kumar Tripathi, Swapna Chaudhuri.
  Journal of Cellular Physiology. March 24, 2017
  Combating gliomagenic global immunosuppression is one of the emerging key for improving prognosis in malignant glioma. Apoptosis plays a pivotal role within the adult hematopoietic system particularly in regulating the cells of immune system. Gliomagenic regulation of apoptotic mediators within bone marrow milieu has not been elucidated. We previously demonstrated that administration of membrane glycopeptides T11 target structure (T11TS) not only rejuvenate bone marrow hematopoietic stem cells (BMHSCs) from glioma mediated hibernation by inhibiting gliomagenic overexpression of Ang‐1/Tie‐2 but also stimulate glioma mediated diminution of expression CD34, c‐kit, and Sca‐1 markers. In the present study, we investigated the impact of glioma on apoptotic signaling cascades of BMHSCs and consequences following T11TS therapy. Bone marrow smear and Annexin V staining confirm gliomagenic acceleration of apoptotic fate of BMHSCs whereas T11TS treatment in glioma‐bearing rats disrupted apoptosis of BMHSCs. Flowcytometry, immunoblotting, and immunofluorescence imagining results revealed multi potent T11TS not only significantly downregulates gliomagenic overexpression of Fas, Fas L, Bid, and caspase‐8, the pro‐apoptotic extrinsic mediators but also strongly inhibits cytosolic release of cytochrome‐c, Apf‐1, and Bax to deactivate gliomagenic caspase‐9, 3 the key intrinsic apoptotic mediators followed by up modulation of anti‐apoptotic Bcl‐2 in glioma associated HSCs. T11TS is also able to diminish the perforin‐granzyme B mediated apoptotic verdict of BMHSCs during gliomagenesis. The anti‐apoptotic action of T11TS on glioma associated BMHSCs provide a crucial insight into how T11TS exerts its immunomodulatory action against glioma mediated immune devastation. T11TS inhibits gliomagenic acceleration of premature apoptosis of bone marrow hematopoietic stem cells by repressing extrinsic, intrinsic, and granzyme B mediated apoptogenic cascades. Flowcytometry, immunoblotting experiments and immunofluorescence imagining results revealed multi potent T11TS significantly downregulates gliomaogenic overexpression of the key apoptotic mediators of all the above three pathways.

  March 24, 2017   doi: 10.1002/jcp.25874   open full text
• Mild Aerobic Exercise Training Hardly Affects the Diaphragm of mdx Mice.
  Giuseppe Morici, Monica Frinchi, Alessandro Pitruzzella, Valentina Di Liberto, Rosario Barone, Andrea Pace, Valentina Di Felice, Natale Belluardo, Francesco Cappello, Giuseppa Mudò, Maria R. Bonsignore.
  Journal of Cellular Physiology. March 24, 2017
  In the mdx mice model of Duchenne Muscular Dystrophy (DMD), mild endurance exercise training positively affected limb skeletal muscles, whereas few and controversial data exist on the effects of training on the diaphragm. The diaphragm was examined in mdx (C57BL/10ScSn‐Dmdmdx) and wild‐type (WT, C57BL/10ScSc) mice under sedentary conditions (mdx‐SD, WT‐SD) and during mild exercise training (mdx‐EX, WT‐EX). At baseline, and after 30 and 45 days (training: 5 d/wk for 6 weeks), diaphragm muscle morphology and Cx39 protein were assessed. In addition, tissue levels of the chaperonins Hsp60 and Hsp70 and the p65 subunit of nuclear factor‐kB (NF‐kB) were measured in diaphragm, gastrocnemius, and quadriceps in each experimental group at all time points. Although morphological analysis showed unchanged total area of necrosis/regeneration in the diaphragm after training, there was a trend for larger areas of regeneration than necrosis in the diaphragm of mdx‐EX compared to mdx‐SD mice. However, the levels of Cx39, a protein associated with active regeneration in damaged muscle, were similar in the diaphragm of mdx‐EX and mdx‐SD mice. Hsp60 significantly decreased at 45 days in the diaphragm, but not in limb muscles, in both trained and sedentary mdx compared to WT mice. In limb muscles, but not in the diaphragm, Hsp70 and NF‐kB p65 levels were increased in mdx mice irrespective of training at 30 and 45 days. Therefore, the diaphragm of mdx mice showed little inflammatory and stress responses over time, and appeared hardly affected by mild endurance training. J. Cell. Physiol. 232: 2044–2052, 2017. © 2016 Wiley Periodicals, Inc. Differently from limb muscles, the diaphragm of mdx mice showed no evidence of decreased muscle damage over time, as measured by Cx39 protein, in exercised compared to sedentary mice. Such findings were associated with progressive decrease of Hsp60 expression, and unchanged Hsp70 and NF‐kB expression in the diaphragm, irrespective of training. The data suggest that mild exercise training hardly affects the diaphragm muscle in the mdx mice model of Duchenne Muscular Dystrophy (DMD).

  March 24, 2017   doi: 10.1002/jcp.25573   open full text
• Mimicking exercise in three‐dimensional bioengineered skeletal muscle to investigate cellular and molecular mechanisms of physiological adaptation.
  Andreas M. Kasper, Daniel C. Turner, Neil R. W. Martin, Adam P. Sharples.
  Journal of Cellular Physiology. March 21, 2017
  Bioengineering of skeletal muscle in vitro in order to produce highly aligned myofibres in relevant three dimensional (3D) matrices have allowed scientists to model the in vivo skeletal muscle niche. This review discusses essential experimental considerations for developing bioengineered muscle in order to investigate exercise mimicking stimuli. We identify current knowledge for the use of electrical stimulation and co‐culture with motor neurons to enhance skeletal muscle maturation and contractile function in bioengineered systems in vitro. Importantly, we provide a current opinion on the use of acute and chronic exercise mimicking stimuli (electrical stimulation and mechanical overload) and the subsequent mechanisms underlying physiological adaptation in 3D bioengineered muscle. We also identify that future studies using the latest bioreactor technology, providing simultaneous electrical and mechanical loading and flow perfusion in vitro, may provide the basis for advancing knowledge in the future. We also envisage, that more studies using genetic, pharmacological, and hormonal modifications applied in human 3D bioengineered skeletal muscle may allow for an enhanced discovery of the in‐depth mechanisms underlying the response to exercise in relevant human testing systems. Finally, 3D bioengineered skeletal muscle may provide an opportunity to be used as a pre‐clinical in vitro test‐bed to investigate the mechanisms underlying catabolic disease, while modelling disease itself via the use of cells derived from human patients without exposing animals or humans (in phase I trials) to the side effects of potential therapies. This review discusses the current understanding, advances, and future directions for the stimulation of three‐dimensional bioengineered skeletal muscle to investigate the mechanisms of physiological adaptation to exercise.

  March 21, 2017   doi: 10.1002/jcp.25840   open full text
• HMGB1 down‐regulation mediates terameprocol vascular anti‐proliferative effect in experimental pulmonary hypertension.
  Rita Nogueira‐Ferreira, Manuel J. Ferreira‐Pinto, Ana Filipa Silva, Rui Vitorino, Joana Justino, Raquel Costa, Daniel Moreira‐Gonçalves, Jean‐François Quignard, Thomas Ducret, Jean‐Pierre Savineau, Adelino F. Leite‐Moreira, Rita Ferreira, Tiago Henriques‐Coelho.
  Journal of Cellular Physiology. March 09, 2017
  Pulmonary arterial hypertension (PAH) is a progressive disease with a poor prognosis. Pulmonary artery smooth muscle cells (PASMCs) play a crucial role in PAH pathophysiology, displaying a hyperproliferative, and apoptotic‐resistant phenotype. In the present study, we evaluated the potential therapeutic role of terameprocol (TMP), an inhibitor of cellular proliferation and promoter of apoptosis, in a well‐established pre‐clinical model of PAH induced by monocrotaline (MCT) and studied the biological pathways modulated by TMP in PASMCs. Wistar rats injected with MCT or saline (SHAM group) were treated with TMP or vehicle. On day 21 after injection, we assessed bi‐ventricular hemodynamics and cardiac and pulmonary morphometry. The effects of TMP on PASMCs were studied in a primary culture isolated from SHAM and MCT‐treated rats, using an iTRAQ‐based proteomic approach to investigate the molecular pathways modulated by this drug. In vivo, TMP significantly reduced pulmonary and cardiac remodeling and improved cardiac function in PAH. In vitro, TMP inhibited proliferation and induced apoptosis of PASMCs. A total of 65 proteins were differentially expressed in PASMCs from MCT rats treated with TMP, some of which involved in the modulation of transforming growth factor beta pathway and DNA transcription. Anti‐proliferative effect of TMP seems to be explained, at least in part, by the down‐regulation of the transcription factor HMGB1. Our findings support the beneficial role of TMP in PAH and suggest that it may be an effective therapeutic option to be considered in the clinical management of PAH. TMP‐reduced pulmonary and cardiac remodeling and hemodynamic features of PAH. Its vascular anti‐proliferative effect seems to be mediated by the protein HMGB1. TMP may be an effective therapeutic option in the clinical management of PAH.

  March 09, 2017   doi: 10.1002/jcp.25763   open full text
• Cytokine networks and their association with Helicobacter pylori infection in gastric carcinoma.
  Vahid Bagheri, Bahram Memar, Amir Abbas Momtazi, Amirhossein Sahebkar, Mehran Gholamin, Mohammad Reza Abbaszadegan.
  Journal of Cellular Physiology. March 07, 2017
  Cytokine networks as dynamic networks are pivotal aspects of tumor immunology, especially in gastric cancer (GC), in which infection, inflammation, and antitumor immunity are key elements of disease progression. In this review, we describe functional roles of well‐known GC‐modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)‐induced inflammation in GC and discuss how HP‐induced chronic inflammation can lead to the induction of stem cell hyperplasia, morphological changes in gastric mucosa and GC development. In this review, we describe functional roles of well‐known gastric cancer (GC)‐modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)‐induced inflammation in GC and discuss how HP‐induced chronic inflammation can lead to GC.

  March 07, 2017   doi: 10.1002/jcp.25822   open full text
• 2‐Methoxyestradiol Impacts on Amino Acids‐mediated Metabolic Reprogramming in Osteosarcoma Cells by Interaction with NMDA Receptor.
  Magdalena Gorska‐Ponikowska, Ugo Perricone, Alicja Kuban‐Jankowska, Giosue Lo Bosco, Giampaolo Barone.
  Journal of Cellular Physiology. March 06, 2017
  Deregulation of serine and glycine metabolism, have been identified to function as metabolic regulators in supporting tumor cell growth. The role of serine and glycine in regulation of cancer cell proliferation is complicated, dependent on concentrations of amino acids and tissue‐specific. D‐serine and glycine are coagonists of N‐methyl‐D‐aspartate receptor subunit GRIN1. Importantly, NMDA receptors are widely expressed in cancer cells and play an important role in regulation of cell death, proliferation and metabolism of numerous malignancies. The aim of the present work was to associate the metabolism of glycine and D‐serine with the anticancer activity of 2‐methoxyestradiol. 2‐methoxyestradiol is a potent anticancer agent but also a physiological 17β‐ estradiol metabolite. In the study we have chosen two malignant cell lines expressing functional GRIN1 receptors, i.e. osteosarcoma 143B and breast cancer MCF7. We used MTS assay, migration assay, flow cytometric analyses, western blotting and immunoprecipitation techniques as well as molecular modeling studies. We have demonstrated the extensive crosstalk between the deregulated metabolic network and cancer cell signaling. Herein, we observed an anticancer effect of high concentrations of glycine and D‐serine in osteosarcoma cells. In contrast, the amino acids when used at low, physiological concentrations induced the proliferation and migration of osteosarcoma and breast cancer cells. Importantly, the pro‐cancergogenic effects of both glycine and D‐serine where abrogated by the usage of 2‐methoxyestradiol at both physiological and pharmacological relevant concentrations. The obtained data confirmed that 2‐methoxyestradiol may be a physiological anticancer molecule. This article is protected by copyright. All rights reserved

  March 06, 2017   doi: 10.1002/jcp.25888   open full text
• Reciprocal regulation between βTrCP and Smurf1 suppresses proliferative capacity of liver cancer cells.
  Yue Zhang, Wenhua Wang, Si Cai, Yan Chen, Qinwan Wang, Qiuhui Pan, Fenyong Sun, Jiayi Wang.
  Journal of Cellular Physiology. March 06, 2017
  We previously reported that both the ubiquitin E3 ligases βTrCP (beta‐transducin repeat‐containing E3 ubiquitin protein ligase) and Smurf1 (SMAD‐specific E3 ubiquitin protein ligase 1) play similar antitumorigenic roles in liver cancer cells. However, whether and how they are reciprocally regulated remains elusive. Here, we show that βTrCP interacts with Smurf1 through the 7 × tryptophan (W) aspartic acid (D)(WD) 40 and the region homologous to the E6‐AP carboxyl terminus (HECT) domains, which are the E3 ligase domains of βTrCP and Smurf1, respectively. The E3 ligase domains of βTrCP and Smurf1 are also critical for maintaining the protein expressions of Smurf1 and βTrCP. Moreover, a positive correlation between βTrCP and Smurf1 was also revealed by tissue microarray analysis, indicating that this relationship might be important in liver cancer. Further, we found that Smurf1 increases the protein stability of βTrCP, possibly by reducing autoubiquitination of βTrCP, and vice versa. Interestingly, such effects depended on the presence of E3 ligase domains. Importantly, depletion of Smurf1‐ or βTrCP‐enhanced proliferative capacity of liver cancer cells could be partially reversed by overexpression of wild‐type βTrCP or Smurf1 but not their E3 ligase‐dead mutants. Collectively, a reciprocal post‐translational regulation between βTrCP and Smurf1 has been uncovered in this study. Simultaneous enhancement of βTrCP and Smurf1 functions might be helpful in the treatment of liver cancer. In the present study, we identified a close interaction between βTrCP and Smurf1, uncovering a reciprocal post‐translational regulation between RING and HECT E3s in liver cancer cells. Simultaneous stimulation of βTrCP and Smurf1 functions may help to conquer liver cancer.

  March 06, 2017   doi: 10.1002/jcp.25780   open full text
• TUDCA: An Agonist of the Bile Acid Receptor GPBAR1/TGR5 With Anti‐Inflammatory Effects in Microglial Cells.
  Natalia Yanguas‐Casás, M. Asunción Barreda‐Manso, Manuel Nieto‐Sampedro, Lorenzo Romero‐Ramírez.
  Journal of Cellular Physiology. March 03, 2017
  Bile acids are steroid acids found in the bile of mammals. The bile acid conjugate tauroursodeoxycholic acid (TUDCA) is neuroprotective in different animal models of stroke and neurological diseases. We have previously shown that TUDCA has anti‐inflammatory effects on glial cell cultures and in a mouse model of acute neuroinflammation. We show now that microglial cells (central nervous system resident macrophages) express the G protein‐coupled bile acid receptor 1/Takeda G protein‐coupled receptor 5 (GPBAR1/TGR5) in vivo and in vitro. TUDCA binding to GPBAR1/TGR5 caused an increase in intracellular cAMP levels in microglia that induced anti‐inflammatory markers, while reducing pro‐inflammatory ones. This anti‐inflammatory effect of TUDCA was inhibited by small interference RNA for GPBAR1/TGR5 receptor, as well as by treatment with a protein kinase A (PKA) inhibitor. In the mouse model of acute neuroinflammation, treating the animals with TUDCA was clearly anti‐inflammatory. TUDCA biased the microglial phenotype in vivo and in vitro toward the anti‐inflammatory. The bile acid receptor GPBAR1/TGR5 could be a new therapeutic target for pathologies coursing with neuroinflammation and microglia activation, such as traumatic brain injuries, stroke, or neurodegenerative diseases. TUDCA and other GPBAR1/TGR5 agonists need to be further investigated, to determine their potential in attenuating the neuropathologies associated with microglia activation. J. Cell. Physiol. 232: 2231–2245, 2017. © 2016 Wiley Periodicals, Inc. We provide evidence that tauroursodeoxycholic acid (TUDCA) exerts anti‐inflammatory effects through its binding to GPBAR1/TGR5, and the consequent increase in intracellular cAMP levels in microglia. Besides, TUDCA treatment biased microglia towards the anti‐inflammatory phenotype under pro‐inflammatory conditions in vitro and in vivo. TUDCA and other GPBAR1/TGR5 agonists are potential therapeutic targets for the treatment of neuropathologies associated with microglia activation.

  March 03, 2017   doi: 10.1002/jcp.25742   open full text
• Association between AXL, Hippo Transducers, and Survival Outcomes in Male Breast Cancer.
  Anna Di Benedetto, Marcella Mottolese, Francesca Sperati, Cristiana Ercolani, Luigi Di Lauro, Laura Pizzuti, Patrizia Vici, Irene Terrenato, Abeer M. Shaaban, Matthew P. Humphries, Sreekumar Sundara‐Rajan, Maddalena Barba, Valerie Speirs, Ruggero De Maria, Marcello Maugeri‐Saccà.
  Journal of Cellular Physiology. March 03, 2017
  Male breast cancer (MBC) is an uncommon malignancy. We have previously reported that the expression of the Hippo transducers TAZ/YAP and their target CTGF was associated with inferior survival in MBC patients. Preclinical evidence demonstrated that Axl is a transcriptional target of TAZ/YAP. Thus, we herein assessed AXL expression to further investigate the significance of active TAZ/YAP‐driven transcription in MBC. For this study, 255 MBC samples represented in tissue microarrays were screened for AXL expression, and 116 patients were included. The association between categorical variables was verified by the Pearson's Chi‐squared test of independence (2‐tailed) or the Fisher Exact test. The relationship between continuous variables was tested with the Pearson's correlation coefficient. The Kaplan–Meier method was used for estimating survival curves, which were compared by log‐rank test. Factors potentially impacting 10‐year and overall survival were verified in Cox proportional regression models. AXL was positively associated with the TAZ/CTGF and YAP/CTGF phenotypes (P = 0.001 and P = 0.002, respectively). Patients with TAZ/CTGF/AXL‐ or YAP/CTGF/AXL‐expressing tumors had inferior survival compared with non‐triple‐positive patients (log rank P = 0.042 and P = 0.048, respectively). The variables TAZ/CTGF/AXL and YAP/CTGF/AXL were adverse factors for 10‐year survival in the multivariate Cox models (HR 2.31, 95%CI:1.02–5.22, P = 0.045, and HR 2.27, 95%CI:1.00–5.13, P = 0.050). Nearly comparable results were obtained from multivariate analyses of overall survival. The expression pattern of AXL corroborates the idea of the detrimental role of TAZ/YAP activation in MBC. Overall, Hippo‐linked biomarkers deserve increased attention in this rare disease. J. Cell. Physiol. 232: 2246–2252, 2017. © 2016 Wiley Periodicals, Inc. (1) The expression pattern of AXL corroborates the idea of TAZ/YAP activation in male breast cancer; (2) patients whose tumors had the TAZ/CTGF/AXL and YAP/CTGF/AXL molecular backgrounds experienced adverse survival outcomes compared with their negative counterparts.

  March 03, 2017   doi: 10.1002/jcp.25745   open full text
• Exhaustive Training Leads to Hepatic Fat Accumulation.
  Alisson L. da Rocha, Ana P. Pinto, Giovana R. Teixeira, Bruno C. Pereira, Luciana C. Oliveira, Adriana C. Silva, Gustavo P. Morais, Dennys E. Cintra, José R. Pauli, Adelino S. R. da Silva.
  Journal of Cellular Physiology. March 03, 2017
  Recently, we demonstrated that an overtraining (OT) protocol for mice based on downhill running sessions increased the hepatic phosphorylation of 70‐kDa ribosomal protein S6 kinase 1 (S6K1; Thr389), a downstream target of the mammalian target of rapamycin complex 1 (mTORC1). In liver, the overactivation of the Akt/mTORC1 pathway induces lipogenesis via regulation of the action of sterol regulatory element binding protein‐1 (SREBP‐1) at multiple steps. Herein, we verified the effects of three running OT models with same external load (i.e., the product between intensity and volume of training), but performed in downhill, uphill and without inclination, on the proteins related to the mTORC1 signaling pathway, the protein content of the SREBP‐1, ACC, and FAS, and the morphological characteristics of C57BL/6 mouse livers. In summary, the downhill running‐induced OT model up‐regulated the levels of major proteins of the mTORC1 signaling pathway, the protein levels of SREBP‐1 (p125 precursor) and induced signs of cell swelling accompanied by acute inflammation. The other two OT protocols performed uphill and without inclination did not modulate the most analyzed molecular proteins, but induced hepatic morphological alterations, suggesting an acute pathological adaptation. The three OT models induced hepatic fat accumulation. J. Cell. Physiol. 232: 2094–2103, 2017. © 2016 Wiley Periodicals, Inc. The downhill running‐induced overtraining model increased the levels of major proteins of the mTOR signaling pathway in mouse livers despite high levels of pAMPK (Thr172). The three overtraining models increased the percentage of fat in liver.

  March 03, 2017   doi: 10.1002/jcp.25625   open full text
• Tumor Necrosis Factor‐α Increases Claudin‐1, 4, and 7 Expression in Tubular Cells: Role in Permeability Changes.
  Yasaman Amoozadeh, Qinghong Dan, Shaista Anwer, Hsiao Han Huang, Vanessa Barbieri, Faiza Waheed, Mazharul Maishan, Katalin Szászi.
  Journal of Cellular Physiology. March 03, 2017
  Tumor necrosis factor‐α (TNFα), is a pathogenic cytokine in kidney disease that alters expression of claudins in tubular cells. Previously we showed that in LLC‐PK1 cells TNFα caused a biphasic change in transepithelial resistance (TER) consisting of an early drop and recovery, followed by a late increase. However, the underlying mechanisms and the role of specific claudins in the TER effect remained incompletely understood. Here we sought to define how TNFα affects claudins 1, 4, and 7 in tubular cells and to correlate their changes with the TER effect. We show that TNFα elevates total and surface levels of Cldn‐1, 4, and 7, and increases their mRNA expression through the ERK and JNK pathways. Further, JNK is also important for TNFα‐induced changes in claudin‐2 expression. Continuous monitoring of TER using Electric cell‐substrate impedance sensing (ECIS) reveals that the two phases of the TNFα effect are differently regulated. Specifically, inhibition of the ERK or JNK pathways prevent the late TER increase, but not the early TER effect. Silencing experiments also show that Cldn‐1 is necessary for the early TNFα‐induced TER change, while all three claudins appear to contribute to the late TER increase. In summary, we define a central role for ERK and JNK in TNFα‐induced altered claudin expression and barrier tightening. Together, our current and previous works show that the TNFα‐induced early TER effect requires claudin‐1, while claudin‐2 decrease is a significant mediator of the late TER increase, and elevation in claudin‐1, 4, and 7 contribute to a smaller extent. J. Cell. Physiol. 232: 2210–2220, 2017. © 2016 Wiley Periodicals, Inc. We demonstrate that TNFα elevates levels of claudins 1,4, and 7 and define a central role for ERK and JNK in this effect. Using ECIS we correlate these signalling pathways and claudins with the biphasic TER changes induced by TNFα and show their differential roles in the early and late phases.

  March 03, 2017   doi: 10.1002/jcp.25736   open full text
• Dlx3 and GCM‐1 functionally coordinate the regulation of placental growth factor in human trophoblast‐derived cells.
  Sha Li, Mark S. Roberson.
  Journal of Cellular Physiology. March 01, 2017
  Placental growth factor (PGF) is abundantly expressed by trophoblast cells within human placentae and is important for trophoblast development and placental vascularization. Circulating maternal serum levels of PGF are dynamically upregulated across gestation in normal pregnancies, whereas low circulating levels and placental production of PGF have been implicated in the pathogenesis of preeclampsia and other gestational diseases. However, the underlying molecular mechanism of regulating PGF expression in the human placenta remains poorly understood. In this study, we demonstrated that transcription factors Distal‐less 3 (DLX3) and Glial cell missing‐1 (GCM1) were both sufficient and required for PGF expression in human trophoblast‐derived cells by overexpression and knockdown approaches. Surprisingly, while DLX3 and GCM1 were both positive regulators of PGF, co‐overexpression of DLX3 and GCM1 led to an antagonist effect on PGF expression on the endogenous gene and a luciferase reporter. Further, deletion and site‐directed mutagenesis studies identified a novel regulatory element on the PGF promoter mediating both DLX3‐ and GCM1‐dependent PGF expression. This regulatory region was also found to be essential for the basal activity of the PGF promoter. Finally, Chromatin‐immunoprecipitation (ChIP) assays revealed colocalization of DLX3 and GCM1 at the identified regulatory region on the PGF promoter. Taken together, our studies provide important insights into intrinsic regulation of human placental PGF expression through the functional coordination of DLX3 and GCM1, and are likely to further the understanding of pathogenesis of PGF dysregulation in preeclampsia and other disease conditions. Our studies examine the role and requirement for the transcription factors Dlx3 and GCM‐1 in the regulation of placental growth factor (PGF) in human placental trophoblast cell lines. Extensive over‐expression and knockdown studies reveal a requirement for both factors; however, the combined action of Dlx3 and GCM‐1 is antagonistic to PGF expression. Our studies identify a novel cis‐acting regulatory element in the proximal region of the PGF gene promoter that binds both factors and is required for the antagonist effects of Dlx3 on GCM‐1.

  March 01, 2017   doi: 10.1002/jcp.25752   open full text
• Tolerogenic probiotics: potential immunoregulators in Systemic Lupus Erythematosus.
  Seyed‐Alireza Esmaeili, Mahmoud Mahmoudi, Amir Abbas Momtazi, Amirhossein Sahebkar, Hassan Doulabi, Maryam Rastin.
  Journal of Cellular Physiology. March 01, 2017
  Probiotics are commensal or nonpathogenic microbes that colonize the gastrointestinal tract and confer beneficial effects on the host through several mechanisms such as competitive exclusion, anti‐bacterial effects, and modulation of immune responses. There is growing evidence supporting the immunomodulatory ability of some probiotics. Several experimental and clinical studies have been shown beneficial effect of some probiotic bacteria, particularly Lactobacillus and Bifidobacteria strains, on inflammatory and autoimmune diseases. Systemic lupus erythematosus (SLE) is an autoimmune disease that is mainly characterized by immune intolerance towards self‐antigens. Some immunomodulatory probiotics have been found to regulate immune responses via tolerogenic mechanisms. Dendritic and T regulatory (Treg) cells, IL‐6, IFN‐γ, IL‐17, and IL‐23 can be considered as the most determinant dysregulated mediators in tolerogenic status. As demonstrated by documented experimental and clinical trials on inflammatory and autoimmune diseases, a number of probiotic bacterial strains can restore tolerance in host through modification of such dysregulated mediators. Since there are limited reports regarding to impact of probiotic supplementation in SLE patients, the preset review was aimed to suggest a number of probiotics bacteria, mainly from Bifidobacteria and Lactobacillus strains that are able to ameliorate immune responses. The aim was followed through literature survey on immunoregulatory probiotics that can restore tolerance and also modulate the important dysregulated pro/anti‐inflammatory cytokines contributing to the pathogenesis of SLE. Since there are limited reports regarding to impact of probiotic supplementation in SLE patients, the preset review was aimed to suggest a number of probiotics bacteria, mainly from Bifidobacteria and Lactobacillus strains that are able to ameliorate immune responses. The aim was followed through literature survey on immunoregulatory probiotics that can restore tolerance and also modulate the important dysregulated pro/anti‐inflammatory cytokines contributing to the pathogenesis of SLE.

  March 01, 2017   doi: 10.1002/jcp.25748   open full text
• Uremic Serum Impairs Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stromal Cells.
  Elena Della Bella, Stefania Pagani, Gianluca Giavaresi, Irene Capelli, Giorgia Comai, Chiara Donadei, Maria Cappuccilli, Gaetano La Manna, Milena Fini.
  Journal of Cellular Physiology. March 01, 2017
  Chronic Kidney Disease‐Mineral and Bone Disorder (CKD‐MBD) is characterized by an increased fracture risk. Bone marrow mesenchymal stromal cells (BMSCs) may be involved in the pathogenesis of bone disease and, in view of their promising potential applications in bone tissue engineering, the effect of uremia on BMSCs regenerative potential represents a central issue. The present study evaluated in vitro the effect of a serum pool from hemodialysis patients on BMSCs to observe its influence on osteogenic differentiation. Besides alterations in spatial organization and cytotoxicity along with hyperproliferation, gene expression analysis suggested an impairment in the osteogenic differentiation. More importantly, Receptor activator of nuclear factor kappa‐B ligand (RANKL) was upregulated with a mild reduction in osteoprotegerin levels. In summary, uremic environment seems to impair BMSCs osteogenic differentiation. Moreover BMSCs themselves may enhance osteoclastogenesis, feasibly contributing to the altered bone remodeling in CKD‐MBD patients. J. Cell. Physiol. 232: 2201–2209, 2017. © 2016 Wiley Periodicals, Inc. BMSCs were cultured in presence of a serum pool from hemodialysis patients (HUS) to study the effect of uremia on BMSC behavior. HUS induced both hyperproliferation and cytotoxicity, with an alteration in the spatial organization of cells. Moreover, osteogenic differentiation was impaired: a reduced mineralization and an altered pattern of gene expression were observed, in particular upregulation of RANKL and reduction in OPG levels.

  March 01, 2017   doi: 10.1002/jcp.25732   open full text
• Impact of Cryopreservation on Caprine Fetal Adnexa Derived Stem Cells and Its Evaluation for Growth Kinetics, Phenotypic Characterization, and Wound Healing Potential in Xenogenic Rat Model.
  Anjali Somal, Irfan A. Bhat, Indu B. , Anuj P. Singh, Bibhudatta S.K. Panda, Perumal A. Desingu, Sriti Pandey, Mukesh K. Bharti, Amar Pal, Guttula Saikumar, Vikash Chandra, Guttula Taru Sharma.
  Journal of Cellular Physiology. March 01, 2017
  This study was conducted to know the impact of cryopreservation on caprine fetal adnexa derived mesenchymal stem cells (MSCs) on the basic stem cell characteristics. Gravid caprine uteri (2–3 months) were collected from local abattoir to derive (amniotic fluid [cAF], amniotic sac [cAS], Wharton's jelly [cWJ], and cord blood [cCB]) MSCs and expanded in vitro. Cells were cryopreserved at 3rd passage (P3) using 10% DMSO. Post‐thaw viability and cellular properties were assessed. Cells were expanded to determine growth kinetics, tri‐lineage differentiation, localization, and molecular expression of MSCs and pluripotency markers; thereafter, these cells were transplanted in the full‐thickness (2 × 2cm2) rat skin wound to determine their wound healing potential. The post‐thaw (pt) growth kinetics study suggested that cWJ MSCs expanded more rapidly with faster population doubling time (PDT) than that of other fetal adnexa MSCs. The relative mRNA expression of surface antigens (CD73, CD90, and CD 105) and pluripotency markers (Oct4, KLF, and cMyc) was higher in cWJ MSCs in comparison to cAS, cAF, and cCB MSCs post‐thaw. The percent wound contraction on 7th day was more than 50% for all the MSC‐treated groups (pre and post‐thaw), against 39.55% in the control group. On day 28th, 99% and more wound contraction was observed in cAF, cAF‐pt, cAS‐pt, cWJ, cWJ‐pt, and cCB, MSCs with better scores for epithelization, neovascularization, and collagen characteristics at a non‐significant level. It is concluded that these MSCs could be successfully cryopreserved without altering their stemness and wound healing properties. J. Cell. Physiol. 232: 2186–2200, 2017. © 2016 Wiley Periodicals, Inc. The present study showed that cWJ MSCs have better pluripotency and post thaw survivability than other fetal adnexa derived MSCs and all the adnexa derived MSCs were found to heal excision wound faster and quality of healed wound was also better than control, indicating that these cells can be cryopreserved and may be used for regenerative cell therapy.

  March 01, 2017   doi: 10.1002/jcp.25731   open full text
• Notch Signaling Participates in TGF‐β‐Induced SOST Expression Under Intermittent Compressive Stress.
  Jeeranan Manokawinchoke, Piyamas Sumrejkanchanakij, Prasit Pavasant, Thanaphum Osathanon.
  Journal of Cellular Physiology. March 01, 2017
  Notch signaling is regulated by mechanical stimuli in various cell types. It has previously been reported that intermittent compressive stimuli enhanced sclerostin (SOST) expression in human periodontal ligament cells (hPDLs) by regulating transforming growth factor‐β (TGF‐β) expression. The aim of the present study was to determine the involvement of Notch signaling in the TGF‐β‐induced SOST expression in hPDLs. Cells were treated with intermittent compressive stress in a computer‐controlled apparatus for 24 h. The mRNA and protein expression of the cells were determined by real‐time polymerase chain reaction and Western blot analysis, respectively. In some experiments, the target signaling pathway was impeded by the addition of a TGF‐β receptor kinase inhibitor (SB431542) or a γ‐secretase inhibitor (DAPT). The results demonstrated that hPDLs under intermittent compressive stress exhibited significantly higher NOTCH2, NOTCH3, HES1, and HEY1 mRNA expression compared with control, indicating that mechanical stress induced Notch signaling. DAPT pretreatment markedly reduced the intermittent stress‐induced SOST expression. The expression of NOTCH2, NOTCH3, HES1, and HEY1 mRNA under compressive stress was significantly reduced after pretreatment with SB431542, coinciding with a reduction in SOST expression. Recombinant human TGF‐β1 enhanced SOST, Notch receptor, and target gene expression in hPDLs. Further, DAPT treatment attenuated rhTGF‐β1‐induced SOST expression. In summary, intermittent compressive stress regulates Notch receptor and target gene expression via the TGF‐β signaling pathway. In addition, Notch signaling participates in TGF‐β‐induced SOST expression in hPDLs. J. Cell. Physiol. 232: 2221–2230, 2017. © 2016 Wiley Periodicals, Inc. Intermittent stress stimulates TGF‐β signaling via an unknown mechanism. TGF‐β, in turn, induces the expression of SOST, NOTCH2, NOTCH3, HES1, and HEY1, whose expression was reduced by SB431542. Inhibition SOST expression, but not that of TGF‐β by DAPT and NOTCH2 shRNA indicate that NOTCH signaling is a downstream participant in the TGF‐β‐induced SOST expression.

  March 01, 2017   doi: 10.1002/jcp.25740   open full text
• Hyaluronic Acid Nanohydrogel Loaded With Quercetin Alone or in Combination to a Macrolide Derivative of Rapamycin RAD001 (Everolimus) as a New Treatment for Hormone‐Responsive Human Breast Cancer.
  Vincenzo Quagliariello, Rosario Vincenzo Iaffaioli, Emilia Armenia, Ottavia Clemente, Manlio Barbarisi, Guglielmo Nasti, Massimiliano Berretta, Alessandro Ottaiano, Alfonso Barbarisi.
  Journal of Cellular Physiology. March 01, 2017
  The aim of this study is based on the evaluation of anticancer, anti‐inflammatory activities, and cellular uptake of hyaluronic acid nanohydrogel of quercetin tested alone and in combination to a macrolide derivative of rapamycin RAD001 (everolimus) on hormone‐responsive breast cancer cell line MCF‐7. Biological investigations were focused on the receptor mediated cellular internalization of the nanohydrogel and its abilities to reduce secretion of several cytokines (IL‐8, IL‐6, IL‐19), VEGF, and metalloproteases (MMP‐2, MMP‐9) under pro‐inflammatory conditions. Nanohydrogel show a CD44 dependent endocytosis with evident time dependent cytoplasmatic accumulation with abilities to reduce secretion of all cytokines of ∼60% compared to untreated cells. Combination of formulated quercetin and everolimus leads to a synergistic cytotoxic effects with a Combination Index of 0.38. These results highlights the importance of synergistic effect of the hyaluronic acid nanohydrogel of quercetin with everolimus in the regulation of human breast cancer cell proliferation and emphasize the antitumor and anti‐inflammatory properties of the nanocarrier. J. Cell. Physiol. 232: 2063–2074, 2017. © 2016 Wiley Periodicals, Inc. Hyaluronic acid nanohydrogel of Quercetin has anti‐anticancer and antinflammatory activity in human breast cancer cells. Combination of Everolimus and hyaluronic acid nanohydrogel of Quercetin leads to synergistic cytotoxicity in MCF‐7 call line.

  March 01, 2017   doi: 10.1002/jcp.25587   open full text
• Deregulated ALG‐2/HEBP2 axis alters microtubule dynamics and mitotic spindle behavior to stimulate cancer development.
  Juan Qin, Yang Yang, Siqi Gao, Yang Liu, Fan Yu, Yunqiang Zhou, Rui Lyu, Min Liu, Xinqi Liu, Dengwen Li, Jun Zhou.
  Journal of Cellular Physiology. March 01, 2017
  Cancer cells are characterized by genomic instability, resulting in the accumulation of mutations that promote cancer progression. One way that genomic instability can arise is through improper regulation of the microtubule cytoskeleton that impacts the function of the mitotic spindle. In this study, we have identified a critical role for the interaction between apoptosis‐linked gene 2 (ALG‐2) and heme‐binding protein 2 (HEBP2) in the above processes. Our data show that the gene copy numbers and mRNA levels for both ALG‐2 and HEBP2 are significantly upregulated in breast and lung cancer. Coexpression of ALG‐2 and HEBP2 markedly increases the cytoplasmic pool of ALG‐2 and alters the subcellular distribution of HEBP2. Our data further reveal that abnormality in the ALG‐2/HEBP2 interaction impairs spindle orientation and positioning during mitosis. In addition, this complex appears to modulate the dynamic properties of microtubules in cancer cells. These finding thus uncover an important function for deregulated ALG‐2/HEBP2 axis in cancer development by influencing microtubule dynamics and spindle behavior, providing novel insight into the etiology and pathogenesis of cancer. In this study, we uncover a previously unrecognized role for deregulated ALG‐2/HEBP2 axis in cancer development. Mechanistically, this is mediated by altered microtubule dynamics and impaired spindle orientation and positioning.

  March 01, 2017   doi: 10.1002/jcp.25754   open full text
• Integrated Analysis and MicroRNA Expression Profiling Identified Seven miRNAs Associated With Progression of Oral Squamous Cell Carcinoma.
  Zhong‐Yi Yan, Zhi‐Qing Luo, Lai‐Jian Zhang, Jia Li, Jia‐Qiang Liu.
  Journal of Cellular Physiology. March 01, 2017
  MicroRNAs have been used as diagnostic and prognostic biomarkers for many cancers including oral squamous cell carcinoma (OSCC). Several studies have been shown that microRNA (miRNA) play important roles during the progression of OSCC. However, the results vary largely in different studies due to different platforms and sample sizes. In this study, we systematically evaluated a large scale of miRNA profiles from current qualified OSCC samples, and further investigated the functions of genes regulated by these key miRNAs as well as the signaling pathways through which these miRNA effect carcinogenesis. Seven key miRNAs were identified, and of which three were significantly upregulated, including hsa‐miR‐21, hsa‐miR‐31, hsa‐miR‐338, and four were downregulated, namely hsa‐miR‐125b, hsa‐miR‐133a, hsa‐miR‐133b, and hsa‐miR‐139. The function enrichment analysis revealed that target genes of upregulated miRNAs were associated with cellular protein metabolic process, macromolecule metabolic process, and TGF‐beta pathway, while the targets of downregulated were enriched in negative regulation of macromolecule biosynthetic process and gene expression, and p53, long‐term potentiation and adherens junction pathways. Transcription factor analysis revealed that there were 67 (51.1%) transcription factors influenced by both up and downregulated miRNAs. In summary, seven key miRNAs were found to play essential role in progression of OSCC, as well as the target genes and transcription factors of these miRNAs. The potential functions of these target genes identified in our study may be profitable to diagnosis and prognostic prediction of OSCC as biomarkers. J. Cell. Physiol. 232: 2178–2185, 2017. © 2016 Wiley Periodicals, Inc. Seven key miRNAs were found to play essential role in progression of OSCC, as well as the target genes and transcription factors of these miRNAs. The potential functions of these target genes identified in our study may be profitable to diagnosis and prognostic prediction of OSCC as biomarkers.

  March 01, 2017   doi: 10.1002/jcp.25728   open full text
• Human Telomerase Reverse Transcriptase (hTERT) Positively Regulates 26S Proteasome Activity.
  Eunju Im, Jong Bok Yoon, Han‐Woong Lee, Kwang Chul Chung.
  Journal of Cellular Physiology. February 28, 2017
  Human telomerase reverse transcriptase (hTERT) is the catalytic subunit of telomerase, an RNA‐dependent DNA polymerase that elongates telomeric DNA. hTERT displays several extra‐telomeric functions that are independent of its telomere‐regulatory function, including tumor progression, and neuronal cell death regulation. In this study, we evaluated these additional hTERT non‐telomeric functions. We determined that hTERT interacts with several 19S and 20S proteasome subunits. The 19S regulatory particle and 20S core particle are part of 26S proteasome complex, which plays a central role in ubiquitin‐dependent proteolysis. In addition, hTERT positively regulated 26S proteasome activity independent of its enzymatic activity. Moreover, hTERT enhanced subunit interactions, which may underlie hTERT's ability of hTERT to stimulate the 26S proteasome. Furthermore, hTERT displayed cytoprotective effect against ER stress via the activation of 26S proteasome in acute myeloid leukemia cells. Our data suggest that hTERT acts as a novel chaperone to promote 26S proteasome assembly and maintenance. J. Cell. Physiol. 232: 2083–2093, 2017. © 2016 Wiley Periodicals, Inc. The present study demonstrated that hTERT positively regulates 26S proteasome activity independent of its enzymatic activity. Moreover, hTERT displayed cytoprotective effect against ER stress via the activation of 26S proteasome in acute myeloid leukemia cells. Our data suggest that hTERT acts as a novel chaperone to promote 26S proteasome assembly and maintenance.

  February 28, 2017   doi: 10.1002/jcp.25607   open full text
• Chemoresistance of Lung and Breast Cancer Cells Growing Under Prolonged Periods of Serum Starvation.
  Juan Sebastian Yakisich, Rajkumar Venkatadri, Neelam Azad, Anand Krishnan V. Iyer.
  Journal of Cellular Physiology. February 28, 2017
  The efficacy of chemotherapy is hindered by both tumor heterogeneity and acquired or intrinsic multi‐drug resistance caused by the contribution of multidrug resistance proteins and stemness‐associated prosurvival markers. Therefore, targeting multi‐drug resistant cells would be much more effective against cancer. In this study, we characterized the chemoresistance properties of adherent (anchorage‐dependent) lung H460 and breast MCF‐7 cancer cells growing under prolonged periods of serum starvation (PPSS). We found that under PPSS, both cell lines were highly resistant to Paclitaxel, Colchicine, Hydroxyurea, Obatoclax, Wortmannin, and LY294002. Levels of several proteins associated with increased stemness such as Sox2, MDR1, ABCG2, and Bcl‐2 were found to be elevated in H460 cells but not in MCF‐7 cells. While pharmacological inhibition of either MDR1, ABCG2, Bcl‐2 with Verapamil, Sorafenib, or Obatoclax, respectively decreased the levels of their target proteins under routine culture conditions as expected, such inhibition did not reverse PX resistance in PPSS conditions. Paradoxically, treatment with inhibitors in serum‐starved conditions produced an elevation of their respective target proteins. In addition, we found that Digitoxin, an FDA approved drug that decrease the viability of cancer cells growing under PPSS, downregulates the expression of Sox2, MDR1, phospho‐ AKT, Wnt5a/b, and β‐catenin. Our data suggest that PPSS‐induced chemoresistance is the result of extensive rewiring of intracellular signaling networks and that multi‐resistance can be effectively overcome by simultaneously targeting multiple targets of the rewired network. Furthermore, our PPSS model provides a simple and useful tool to screen drugs for their ability to target multiple pathways of cancer resistance. J. Cell. Physiol. 232: 2033–2043, 2017. © 2016 Wiley Periodicals, Inc. Cancer cells growing under prolonged periods of serum starvation (PPSS) become multidrug resistant. Under PPSS cancer cells respond aberrantly and paradoxically to some anticancer drugs likely by rewire signaling pathway networks.

  February 28, 2017   doi: 10.1002/jcp.25514   open full text
• Isolation, identification, and characterization of cancer stem cells: A review.
  Mohammad Reza Abbaszadegan, Vahid Bagheri, Mahya Shariat Razavi, Amir Abbas Momtazi, Amirhossein Sahebkar, Mehran Gholamin.
  Journal of Cellular Physiology. February 28, 2017
  Cancer stem cells (CSCs) or tumor‐initiating cells (TICs) as a small subset of neoplastic cells are able to produce a tumor (tumorigenesis), maintain the population of tumorigenic cells (self‐renewal), and generate the heterogeneous cells constructing the entire tumor (pluripotency). The research on stationary and circulating CSCs due to resistance to conventional therapies and inability in complete eradication of cancer is critical for developing novel therapeutic strategies for a more effective reduction in the risk of tumor metastasis and cancer recurrence. This review compiles information about different methods of detection and dissociation, side population, cellular markers, and establishment culture of CSCs, as well as characteristics of CSCs such as tumorigenicity, and signaling pathways associated with self‐renewal and the capability of the same histological tumor regeneration in various cancers. This review compiles information about detection, dissociation, side population, cellular markers, and establishment culture of cancer stem cells, as well as characteristics of CSCs such as tumorigenicity, signaling pathways associated with self‐renewal and the capability of the same histological tumor regeneration in various cancers.

  February 28, 2017   doi: 10.1002/jcp.25759   open full text
• Celastrol Attenuates Cadmium‐Induced Neuronal Apoptosis via Inhibiting Ca2+‐CaMKII‐Dependent Akt/mTOR Pathway.
  Ruijie Zhang, Yu Zhu, Xiaoqing Dong, Beibei Liu, Nana Zhang, Xiaoxue Wang, Lei Liu, Chong Xu, Shile Huang, Long Chen.
  Journal of Cellular Physiology. February 28, 2017
  Cadmium (Cd), an environmental and industrial pollutant, affects the nervous system and consequential neurodegenerative disorders. Recently, we have shown that celastrol prevents Cd‐induced neuronal cell death partially by suppressing Akt/mTOR pathway. However, the underlying mechanism remains to be elucidated. Here, we show that celastrol attenuated Cd‐elevated intracellular‐free calcium ([Ca2+]i) level and apoptosis in neuronal cells. Celastrol prevented Cd‐induced neuronal apoptosis by inhibiting Akt‐mediated mTOR pathway, as inhibition of Akt with Akt inhibitor X or ectopic expression of dominant negative Akt reinforced celastrol's prevention of Cd‐induced phosphorylation of S6K1/4E‐BP1 and cell apoptosis. Furthermore, chelating intracellular Ca2+ with BAPTA/AM or preventing [Ca2+]i elevation using EGTA potentiated celastrol's repression of Cd‐induced [Ca2+]i elevation and consequential activation of Akt/mTOR pathway and cell apoptosis. Moreover, celastrol blocked Cd‐elicited phosphorylation of CaMKII, and pretreatment with BAPTA/AM or EGTA enhanced celastrol's suppression of Cd‐increased phosphorylation of CaMKII in neuronal cells, implying that celastrol hinders [Ca2+]i‐mediated CaMKII phosphorylation. Inhibiting CaMKII with KN93 or silencing CaMKII attenuated Cd activation of Akt/mTOR pathway and cell apoptosis, and this was strengthened by celastrol. Taken together, these data demonstrate that celastrol attenuates Cd‐induced neuronal apoptosis via inhibiting Ca2+‐CaMKII‐dependent Akt/mTOR pathway. Our findings underscore that celastrol may act as a neuroprotective agent for the prevention of Cd‐induced neurodegenerative disorders. J. Cell. Physiol. 232: 2145–2157, 2017. © 2016 Wiley Periodicals, Inc. Celastrol prevents Cd‐induced elevation of [Ca2+]i, which results in inhibition of CaMKII phosphorylation. This leads to suppression of CaMKII‐dependent Akt/mTOR pathway, thereby preventing Cd‐induced apoptosis in neuronal cells.

  February 28, 2017   doi: 10.1002/jcp.25703   open full text
• Differential Expression of Estrogen Receptor Variants in Response to Inflammation Signals in Human Airway Smooth Muscle.
  Bharathi Aravamudan, Katelyn J. Goorhouse, Ghanashyam Unnikrishnan, Michael A. Thompson, Christina M. Pabelick, John R. Hawse, Y. S. Prakash, Venkatachalem Sathish.
  Journal of Cellular Physiology. February 24, 2017
  The prevalence of asthma is higher in pre‐pubescent and aging males, and in post‐pubertal females, strongly indicating that sex steroids (especially estrogen) may be an important modulator in lung disease. We recently demonstrated that airway smooth muscle (ASM) expresses both alpha and beta forms of the estrogen receptor (ERα and ERβ) in males and females, and that these receptors regulate intracellular [Ca2+] and ASM contractility. Although both ERα and ERβ have multiple splice variants, it is unclear if and how the expression of these variants is modulated under conditions such as chronic inflammation/asthma. In order to test the hypothesis that the differential expression of ERα and ERβ variants contributes to the pathogenesis of asthma, we profiled the expression of various ERα and ERβ genes in asthmatic and inflamed (TNFα‐ or IL‐13‐treated) ASM. Gene expression was assessed at both the mRNA and protein levels in asthmatic ASM cells or non‐asthmatic cells treated with TNFα (20 ng/ml) or IL‐13 (50 ng/ml). We observed marked variation in the expression of ER isoforms in response to inflammatory stimuli, and in non‐asthmatic versus asthmatic ASM. Changes in protein levels of ERα and ERβ corresponded with the observed differential mRNA patterns. Pharmacological studies implicate cytosolic (p42/44 MAPK and PI3 K) and nuclear (NFκB, STAT6, and AP‐1) signaling pathways as putative mechanisms that mediate and/or regulate effects of inflammation on ER expression. We conclude that variations in ASM ER expression profiles occur with inflammation and that ER variants could contribute to estrogen signaling in airway diseases such as asthma. J. Cell. Physiol. 232: 1754–1760, 2017. © 2016 Wiley Periodicals, Inc.

  February 24, 2017   doi: 10.1002/jcp.25674   open full text
• Cytoplasmic Localization of RUNX3 via Histone Deacetylase‐Mediated SRC Expression in Oxidative‐Stressed Colon Cancer Cells.
  Kyoung Ah Kang, Mei Jing Piao, Yea Seong Ryu, Young Hee Maeng, Jin Won Hyun.
  Journal of Cellular Physiology. February 21, 2017
  Runt domain transcription factor 3 (RUNX3) is a transcription factor that functions as a tumor suppressor. RUNX3 is frequently inactivated by epigenetic silencing or its protein mislocalization (cytoplasmic localization) in many cancer types. This study investigated whether oxidative stress induces redistribution of RUNX3 from the nucleus to the cytoplasm. The cytoplasmic localization of RUNX3 was associated with oxidative stress‐induced RUNX3 phosphorylation at tyrosine residues via SRC activation. Moreover, oxidative stress increased expression of histone deacetylases (HDACs). RUNX3 phosphorylation and SRC expression induced by oxidative stress were inhibited by knockdown of HDAC1, restoring the nuclear localization of RUNX3 under oxidative stress. In conclusion, these results demonstrate that HDAC1‐ and SRC‐mediated phosphorylation of RUNX3 induced by oxidative stress is associated with the cytoplasmic localization of RUNX3 and can lead to RUNX3 inactivation and carcinogenesis. J. Cell. Physiol. 232: 1914–1921, 2017. © 2016 Wiley Periodicals, Inc. RUNX3 cytoplasmic localization induced by oxidative stress in colon cancer cells. Oxidative stress induces HDAC1 expression and then activates SRC. Activated SRC induces expression of phospho‐RUNX3. Phospho‐RUNX3 is retained in the cytoplasm, leading to degradation of cytoplasmic RUNX3 by the proteasome system and finally resulting carcinogenesis.

  February 21, 2017   doi: 10.1002/jcp.25746   open full text
• Na+/H+ exchanger NHE1 and NHE2 have opposite effects on migration velocity in rat gastric surface cells.
  Anja Paehler vor der Nolte, Giriprakash Chodisetti, Zhenglin Yuan, Florian Busch, Brigitte Riederer, Min Luo, Yan Yu, Manoj B. Menon, Andreas Schneider, Renata Stripecke, Katerina Nikolovska, Sunil Yeruva, Ursula Seidler.
  Journal of Cellular Physiology. February 21, 2017
  Following superficial injury, neighbouring gastric epithelial cells close the wound by rapid cell migration, a process called epithelial restitution. Na+/H+ exchange (NHE) inhibitors interfere with restitution, but the role of the different NHE isoforms expressed in gastric pit cells has remained elusive. The role of the basolaterally expressed NHE1 (Slc9a1) and the presumably apically expressed NHE2 (Slc9a2) in epithelial restitution was investigated in the nontransformed rat gastric surface cell line RGM1. Migration velocity was assessed by loading the cells with the fluorescent dye DiR and following closure of an experimental wound over time. Since RGM1 cells expressed very low NHE2 mRNA and have low transport activity, NHE2 was introduced by lentiviral gene transfer. In medium with pH 7.4, RGM1 cells displayed slow wound healing even in the absence of growth factors and independently of NHE activity. Growth factors accelerated wound healing in a partly NHE1‐dependent fashion. Preincubation with acidic pH 7.1 stimulated restitution in a NHE1‐dependent fashion. When pH 7.1 was maintained during the restitution period, migratory speed was reduced to ∼10% of the speed at pH 7,4, and the residual restitution was further inhibited by NHE1 inhibition. Lentiviral NHE2 expression increased the steady‐state pHi and reduced the restitution velocity after low pH preincubation, which was reversible by pharmacological NHE2 inhibition. The results demonstrate that in RGM1 cells, migratory velocity is increased by NHE1 activation, while NHE2 activity inhibit this process. A differential activation of NHE1 and NHE2 may therefore, play a role in the initiation and completion of the epithelial restitution process. Genetic deletion of the Na+/H+ exchanger NHE2 leads to compromised wound healing in the gastrointestinal tract of mice. We studied the roles of NHE1 and NHE2 in gastrointestinal wound healing in vitro, and while NHE1 activity supported wound healing velocity, NHE2/GFP‐overexpression dramatically reduced wound healing velocity compared to GFP expression alone. This effect was abolished by a NHE2‐ but not a NHE1 inhibitory concentration of HOE642. Thus, NHE1 and NHE2 may be important for the initiation and the resolution of gastrointestinal wound healing.

  February 21, 2017   doi: 10.1002/jcp.25758   open full text
• Activin‐SMAD signaling is required for maintenance of porcine iPS cell self‐renewal through upregulation of NANOG and OCT4 expression.
  Fan Yang, Ning Wang, Yaxian Wang, Tong Yu, Huayan Wang.
  Journal of Cellular Physiology. February 21, 2017
  Porcine induced pluripotent stem cells (piPSCs) retain the enormous potential for farm animal reproduction and translational medicine, and have been reported by many laboratories worldwide. Some piPSC lines were bFGF‐dependence and showed mouse EpiSC‐like morphology; other lines were LIF‐dependence and showed mouse ESC‐like morphology. Metastable state of piPSC line that required both LIF and bFGF was also reported. Because bona fide pig embryonic stem cells were not available, uncovering piPSC state‐specific regulatory circuitries was the most important task. In this study, we explored the function of Activin‐SMAD signaling pathway and its downstream activated target genes in piPSCs. Transcriptome analysis showed that genes involved in Activin‐SMAD signaling pathway were evidently activated during porcine somatic cell reprogramming, regardless piPSCs were LIF‐ or bFGF‐dependent. Addition of Activin A and overexpression of SMAD2/3 significantly promoted expressions of porcine NANOG and OCT4, whereas inhibition of Activin‐SMAD signaling by SB431542 and SMAD7 reduced NANOG and OCT4 expressions, and induced piPSCs differentiation exiting from pluripotent state. Our data demonstrate that activation of Activin‐SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self‐renewal in porcine pluripotent stem cells. Our data demonstrate that activation of Activin‐SMAD signaling pathway by addition of Activin A in culture medium is necessary for maintenance of self‐renewal in porcine pluripotent stem cells.

  February 21, 2017   doi: 10.1002/jcp.25747   open full text
• LCN2 overexpression in bone enhances the hematopoietic compartment via modulation of the bone marrow microenvironment.
  Delfina Costa, Elisa Principi, Edoardo Lazzarini, Fiorella Descalzi, Ranieri Cancedda, Patrizio Castagnola, Sara Tavella.
  Journal of Cellular Physiology. February 21, 2017
  Lipocalin‐2 (LCN2) is a member of the lipocalin family whose expression is modulated in several conditions, including cell differentiation, innate immunity, stress, and cancer. Although it is known that it is expressed in bone, its function in this tissue remains poorly studied. To this end, we took advantage of transgenic mice lines that expressed LCN2 driven by a bone specific type I collagen (LCN2‐Tg). In the bone marrow (BM) of LCN2‐Tg mice we observed an increased number of phenotypically long‐term hematopoietic stem cells (LT‐HSC) that also displayed a higher proliferation rate compared to wild‐type controls (Wt). Furthermore, hematopoietic progenitor cells, obtained from LCN2‐Tg BM showed an increased clonogenic capacity compared to those obtained from LCN2‐Tg spleen, a higher concentration of serum erythropoietin and a higher number of mature erythrocytes in the peripheral blood of old LCN2‐Tg animals compared to aged‐matched wt. The findings of a combined increase in the BM of the LCN2‐Tg mice of SDF‐1, SCF, and TIMP‐1 levels along with the reduction of both MMP‐9 activity and cathepsin K concentration may explain the observed effects on the HSC compartment. This study shows that LCN2 overexpression in bones modifies the BM microenvironment via modulation of the expression of key secreted factors and cytokines, which in turn regulate the HSC niche behavior enhancing both HSC homing in young mice and erythrocytes production in older mice. The overexpression of LCN2 in bone stimulates the expansion of the HSC by inducing pro‐retention factors and inhibiting pro‐egress signals for hematopoietic stem cells.

  February 21, 2017   doi: 10.1002/jcp.25755   open full text
• STIM‐1 and ORAI‐1 channel mediate angiotensin‐II‐induced expression of Egr‐1 in vascular smooth muscle cells.
  Estelle R. Simo‐Cheyou, Ju Jing Tan, Ryszard Grygorczyk, Ashok K. Srivastava.
  Journal of Cellular Physiology. February 21, 2017
  An upregulation of Egr‐1 expression has been reported in models of atherosclerosis and intimal hyperplasia and, various vasoactive peptides and growth promoting stimuli have been shown to induce the expression of Egr‐1 in vascular smooth muscle cells (VSMC). Angiotensin‐II (Ang‐II) is a key vasoactive peptide that has been implicated in the pathogenesis of vascular diseases. Ang‐II elevates intracellular Ca2+ through activation of the store‐operated calcium entry (SOCE) involving an inositol‐3‐phosphate receptor (IP3R)‐coupled depletion of endoplasmic reticular Ca2+ and a subsequent activation of the stromal interaction molecule 1 (STIM‐1)/Orai‐1 complex. However, the involvement of IP3R/STIM‐1/Orai‐1‐Ca2+‐dependent signaling in Egr‐1 expression in VSMC remains unexplored. Therefore, in the present studies, we have examined the role of Ca2+ signaling in Ang‐II‐induced Egr‐1 expression in VSMC and investigated the contribution of STIM‐1 or Orai‐1 in mediating this response. 2‐aminoethoxydiphenyl borate (2‐APB), a dual non‐competitive antagonist of IP3R and inhibitor of SOCE, decreased Ang‐II‐induced Ca2+ release and attenuated Ang‐II‐induced enhanced expression of Egr‐1 protein and mRNA levels. Egr‐1 upregulation was also suppressed following blockade of calmodulin and CaMKII. Furthermore, RNA interference‐mediated depletion of STIM‐1 or Orai‐1 attenuated Ang‐II‐induced Egr‐1 expression as well as Ang‐II‐induced phosphorylation of ERK1/2 and CREB. In addition, siRNA‐induced silencing of CREB resulted in a reduction in the expression of Egr‐1 stimulated by Ang‐II. In summary, our data demonstrate that Ang‐II‐induced Egr‐1 expression is mediated by STIM‐1/Orai‐1/Ca2+‐dependent signaling pathways in A‐10 VSMC. Angiotensin‐II‐induced intracellular calcium release depletes the endoplasmic reticulum store, resulting in the activation of the stromal interaction molecule 1 (STIM1) known to cooperate with Orai‐1 transmembrane channel in mediating store‐operated calcium entry (SOCE) in the cells. In this study, we demonstrate that intact SOCE and unaltered expression of STIM‐1 and Orai‐1, in association with an involvement of calcium signaling molecules such as calmodulin and CaMKII, are required for Ang‐II‐dependent induction of the early growth response protein‐1 in VSMC. We also provide evidence of the critical role played by ERK1/2 and CREB activation in mediating this response.

  February 21, 2017   doi: 10.1002/jcp.25810   open full text
• The Role of Cardiac Myosin Binding Protein C3 in Hypertrophic Cardiomyopathy‐Progress and Novel Therapeutic Opportunities.
  Iman A. Mohamed, Navaneethakrishnan T. Krishnamoorthy, Gheyath K. Nasrallah, Sahar I. Da'as.
  Journal of Cellular Physiology. February 16, 2017
  Hypertrophic cardiomyopathy (HCM) is a common autosomal dominant genetic cardiovascular disorder marked by genetic and phenotypic heterogeneity. Mutations in the gene encodes the cardiac myosin‐binding protein C, cMYBPC3 is amongst the various sarcomeric genes that are associated with HCM. These mutations produce mutated mRNAs and truncated cMyBP‐C proteins. In this review, we will discuss the implications and molecular mechanisms involved in MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of HMC. J. Cell. Physiol. 232: 1650–1659, 2017. © 2016 Wiley Periodicals, Inc. This review discusses the implications and the and molecular mechanisms of MYBPC3 different mutations. Further, we will highlight the novel targets that can be developed into potential therapeutics for the treatment of Hypertrophic cardiomyopathy.

  February 16, 2017   doi: 10.1002/jcp.25639   open full text
• Subclinical alteration of the cervical–vaginal microbiome in women with idiopathic infertility.
  Giuseppina Campisciano, Fiorella Florian, Angela D'Eustacchio, David Stanković, Giuseppe Ricci, Francesco De Seta, Manola Comar.
  Journal of Cellular Physiology. February 16, 2017
  Biomarkers have a wide application in research and clinic, they help to choose the correct treatment for diseases. Recent studies, addressing the vaginal microbiome using next generation sequencing (NGS), reported the involvement of bacterial species in infertility. We compared the vaginal microbiome of idiopathic infertile women with that of healthy, including bacterial vaginosis affected women and non‐idiopathic infertile women, to identify bacterial species suitable as biomarkers. Information on microorganisms was obtained from the V3‐16S rDNA sequencing of cervical–vaginal fluids of 96 women using the Ion Torrent platform. Data were processed with QIIME and classified against the Vaginal 16S rDNA Reference Database. The analysis revealed a significant beta‐diversity variation (p < 0.001) between the four groups included in the study. L. iners, L. crispatus, and L. gasseri distinguished idiopathic infertile women from the other groups. In these women, a microbial profile similar to that observed in bacterial vaginosis women has been detected. Our results suggest that the quantitative assessment and identification of specific microorganisms of the cervical–vaginal microflora could increase the accuracy of available tools for the diagnosis of infertility and improve the adoption of therapeutic protocols. Women with repeated failed in vitro fertilization cycles may benefit from microbial screening by NGS. NGS screening could increase the accuracy of available tools for the diagnosis of infertility and improve therapeutic protocols.

  February 16, 2017   doi: 10.1002/jcp.25806   open full text
• Dact1, a Wnt‐Pathway Inhibitor, Mediates Human Mesangial Cell TGF‐β1‐Induced Apoptosis.
  Daniele Pereira Jardim, Paula Cristina Eiras Poço, Alexandre Holthausen Campos.
  Journal of Cellular Physiology. February 16, 2017
  Chronic kidney disease (CKD) is a worldwide public health problem that affects millions of men and women of all ages and racial groups. Loss of mesangial cells (MC) represents an early common feature in the pathogenesis of CKD. Transforming growth factor‐β1 (TGF‐β1) is a key inducer of kidney damage and triggers several pathological changes in renal cells, notably MC apoptosis. However, the mechanism of MC apoptosis induced by TGF‐β1 remains elusive. Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled‐binding antagonist of β‐catenin 1 (Dact1) gene is upregulated by TGF‐β1, inducing MC apoptosis. We also show that the inhibitory effect of Dact1 and TGF‐β1 on the transcriptional activation of the pro‐survival Wnt pathway is the mechanism of death induction. In addition, Dact1 mRNA/protein levels are increased in kidney remnants from 5/6 nephrectomized rats and strongly correlate with TGF‐β1 expression. Together, our results point to Dact1 as a novel element controlling MC survival that is causally related to CKD progression. J. Cell. Physiol. 232: 2104–2111, 2017. © 2016 Wiley Periodicals, Inc. Here, we demonstrate for the first time a novel regulatory pathway in which the disheveled‐binding antagonist of β‐catenin 1 (Dact1) gene is upregulated by TGF‐β1, inducing MC apoptosis.

  February 16, 2017   doi: 10.1002/jcp.25636   open full text
• Identification of MSX1 and DCLK1 as mRNA Biomarkers for Colorectal Cancer Detection Through DNA Methylation Information.
  Ai‐Jun Sun, Hai‐Bo Gao, Gao Liu, Heng‐Fa Ge, Zun‐Ping Ke, Sen Li.
  Journal of Cellular Physiology. February 16, 2017
  Colorectal cancer is the second most deadly malignancy in the United States. However, the currently screening options had their limitation. Novel biomarkers for colorectal cancer detections are necessary to reduce the mortality. The clinical information, mRNA expression levels and DNA methylation information of colorectal cancer were downloaded from TCGA. The patients were separated into training group and testing group based on their platforms for DNA methylation. Beta values of DNA methylation from tumor tissues and normal tissues were utilized to figure out the position that were differentially methylated. The expression levels of mRNA of thirteen genes, whose CpG islands were differentially methylated, were extracted from the RNA‐Seq results from TCGA. The probabilities whether the mRNA was differentially expressed between tumor and normal samples were calculated using Student's t‐test. Logistic regression and decision tree were built for cancer detection and their performances were evaluated by the area under the curve (AUC). Twenty‐four genomic locations were differentially methylated, which could be mapped to eleven genes. Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies. J. Cell. Physiol. 232: 1879–1884, 2017. © 2016 Wiley Periodicals, Inc. Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies.

  February 16, 2017   doi: 10.1002/jcp.25733   open full text
• Mesenchyme Homeobox 2 Enhances Migration of Endothelial Colony Forming Cells Exposed to Intrauterine Diabetes Mellitus.
  Cassandra R. Gohn, Emily K. Blue, BreAnn M. Sheehan, Kaela M. Varberg, Laura S. Haneline.
  Journal of Cellular Physiology. February 16, 2017
  Diabetes mellitus (DM) during pregnancy has long‐lasting implications for the fetus, including cardiovascular morbidity. Previously, we showed that endothelial colony forming cells (ECFCs) from DM human pregnancies have decreased vasculogenic potential. Here, we evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). In human umbilical vein endothelial cells, MEOX2 upregulates cyclin‐dependent kinase inhibitor expression, resulting in increased senescence and decreased proliferation. We hypothesized that dysregulated MEOX2 expression in neonatal ECFCs from DM pregnancies decreases network formation through increased senescence and altered cell cycle progression. Our studies show that nuclear MEOX2 is increased in ECFCs from DM pregnancies. To determine if MEOX2 is sufficient and/or required to induce impaired network formation, MEOX2 was overexpressed and depleted in ECFCs from control and DM pregnancies, respectively. Surprisingly, MEOX2 overexpression in control ECFCs resulted in increased network formation, altered cell cycle progression, and increased senescence. In contrast, MEOX2 knockdown in ECFCs from DM pregnancies led to decreased network formation, while cell cycle progression and senescence were unaffected. Importantly, migration studies demonstrated that MEOX2 overexpression increased migration, while MEOX2 knockdown decreased migration. Taken together, these data suggest that altered migration may be mediating the impaired vasculogenesis of ECFCs from DM pregnancies. While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment. J. Cell. Physiol. 232: 1885–1892, 2017. © 2016 Wiley Periodicals, Inc. Endothelial colony forming cells (ECFCs) from diabetic human pregnancies have decreased vasculogenic potential both in vitro and in vivo. The goal of this study was to evaluate whether the molecular mechanism responsible for this phenotype involves the transcription factor, Mesenchyme Homeobox 2 (MEOX2). While initially believed to be maladaptive, these data suggest that MEOX2 may serve a protective role, enabling increased vessel formation despite exposure to a DM intrauterine environment.

  February 16, 2017   doi: 10.1002/jcp.25734   open full text
• Protective Role of GPER Agonist G‐1 on Cardiotoxicity Induced by Doxorubicin.
  Ernestina M. De Francesco, Carmine Rocca, Francesco Scavello, Daniela Amelio, Teresa Pasqua, Damiano C. Rigiracciolo, Andrea Scarpelli, Silvia Avino, Francesca Cirillo, Nicola Amodio, Maria C. Cerra, Marcello Maggiolini, Tommaso Angelone.
  Journal of Cellular Physiology. February 16, 2017
  The use of Doxorubicin (Dox), a frontline drug for many cancers, is often complicated by dose‐limiting cardiotoxicity in approximately 20% of patients. The G‐protein estrogen receptor GPER/GPR30 mediates estrogen action as the cardioprotection under certain stressful conditions. For instance, GPER activation by the selective agonist G‐1 reduced myocardial inflammation, improved immunosuppression, triggered pro‐survival signaling cascades, improved myocardial mechanical performance, and reduced infarct size after ischemia/reperfusion (I/R) injury. Hence, we evaluated whether ligand‐activated GPER may exert cardioprotection in male rats chronically treated with Dox. 1 week of G‐1 (50 μg/kg/day) intraperitoneal administration mitigated Dox (3 mg/kg/day) adverse effects, as revealed by reduced TNF‐α, IL‐1β, LDH, and ROS levels. Western blotting analysis of cardiac homogenates indicated that G‐1 prevents the increase in p‐c‐jun, BAX, CTGF, iNOS, and COX2 expression induced by Dox. Moreover, the activation of GPER rescued the inhibitory action elicited by Dox on the expression of BCL2, pERK, and pAKT. TUNEL assay indicated that GPER activation may also attenuate the cardiomyocyte apoptosis upon Dox exposure. Using ex vivo Langendorff perfused heart technique, we also found an increased systolic recovery and a reduction of both infarct size and LDH levels in rats treated with G‐1 in combination with Dox respect to animals treated with Dox alone. Accordingly, the beneficial effects induced by G‐1 were abrogated in the presence of the GPER selective antagonist G15. These data suggest that GPER activation mitigates Dox‐induced cardiotoxicity, thus proposing GPER as a novel pharmacological target to limit the detrimental cardiac effects of Dox treatment. J. Cell. Physiol. 232: 1640–1649, 2017. © 2016 Wiley Periodicals, Inc. 1. The potential beneficial effects of GPER activation in Doxorubicin (Dox)‐induced cardiotoxicity was evaluated. 2. In male rat heart, the treatment with the selective GPER ligand G‐1 prevented the increase in TNF‐α, IL‐1β, and LDH plasma levels induced by Dox. In Langendorff perfused rat heart, G‐1 also increased the systolic recovery at the end of ischemia and reduced infarct size and LDH levels triggered by Dox. Moreover, G‐1 attenuated the Dox‐induced myocyte apoptosis, as well as the inhibition of pro‐survival signaling cascades like ERK1/2 and AKT. Next, G‐1 counteracted the down‐regulation of BCL2 as well as the up‐regulation of apoptotic, inflammatory and fibrotic targets, such as BAX, iNOS, COX2, and CTGF induced by Dox. 3. Our study paves the way for further analyzing the role of GPER as pharmacological target in novel therapeutic strategies aimed to prevent the cardiotoxicity exerted by Dox.

  February 16, 2017   doi: 10.1002/jcp.25585   open full text
• Tight Junction Protein Abundance and Apoptosis During Involution of Rat Mammary Glands.
  Claire V. C. Phyn, Kerst Stelwagen, Stephen R. Davis, Christopher D. McMahon, Joanne M. Dobson, Kuljeet Singh.
  Journal of Cellular Physiology. February 16, 2017
  To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement were induced by teat‐sealing with an adhesive for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. In non‐sealed control glands, histological analysis confirmed a lactating phenotype, indicating suckling by pups throughout the experiment. In contrast, alveoli of teat‐sealed glands were distended within 6 h, with maximal luminal size observed by 12 h of non‐suckling. By 18 h following teat‐sealing, an involuting phenotype was observed, indicated by alveolar lumina engorged with milk vesicles and increased leukocytes. Relative to non‐sealed glands, mammary apoptosis was increased in engorged glands 18 h following teat‐sealing. The abundance of ZO‐1 and occludin proteins was decreased in engorged glands by 12 and 18 h, respectively, following teat‐sealing. In contrast, the claudin‐1 22 kDa band was increased by 6 h and peaked at 12–18 h, whereas the 28 kDa band declined by 36 h, relative to controls. There were no temporal changes in ZO‐1, occludin, and claudin‐1 22 kDa proteins within control glands, although there were minor differences in claudin‐1 28 kDa. These data indicate that intramammary milk accumulation due to cessation of milk removal is associated with mammary apoptosis. The apoptotic event is preceded by a rapid loss of abundance of ZO‐1, occludin and an initial increase in claudin‐1. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non‐suckled glands. J. Cell. Physiol. 232: 2075–2082, 2017. © 2016 Wiley Periodicals, Inc. To examine tight junction protein abundance and apoptosis of epithelial cells at the onset of involution in rodent mammary glands, milk accumulation and mammary engorgement was induced by teat‐sealing for 0, 6, 12, 18, 24, and 36 h (n = 6 per group) at peak lactation. The loss of cell–cell communication may initiate involution and apoptosis of mammary epithelial cells and is a localized intramammary event, occurring only in non‐suckled glands.

  February 16, 2017   doi: 10.1002/jcp.25591   open full text
• Postnatal Hyperplasic Effects of ActRIIB Blockade in a Severely Dystrophic Muscle.
  Cory Nielsen, Ross M. Potter, Christopher Borowy, Kimberly Jacinto, Ravi Kumar, C. George Carlson.
  Journal of Cellular Physiology. February 16, 2017
  The efficacy of two ActRIIB ligand‐trapping agents (RAP‐031 and RAP‐435) in treating muscular dystrophy was examined by determining their morphological effects on the severely dystrophic triangularis sterni (TS) muscle of the mdx mouse, a model for Duchenne muscular dystrophy. These agents trap all endogenous ligands to the ActRIIB receptor and thereby block myostatin signaling in a highly selective manner. Short‐term (1 month) and long‐term (3 months) in vivo treatment of 1‐month‐old mdx mice increased myonuclei and fiber cross section (FCS) density but did not alter individual fiber size. Vehicle‐treated mdx mice exhibited age‐dependent increases in myonuclei and FCS density, and age‐dependent reductions in centronucleation that were each enhanced by treatment with RAP‐435. Distributions of FCS area (FCSA) in the mdx TS were 90% identical to those from untreated age‐matched nondystrophic mice and were unaltered by the substantial fiber hyperplasia observed with age and RAP‐435 treatment. These results were inconsistent with injury‐induced fiber regeneration which produces altered FCSA distributions characterized by a distinct class of smaller regenerated fibers. Nondystrophic mice exhibited a constant postnatal density of fiber cross sections and myonuclei, and RAP‐435 treatment of nondystrophic mice increased TS mean FCSA but had no effects on myonuclei or FCS density. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration. J. Cell. Physiol. 232: 1774–1793, 2017. © 2016 Wiley Periodicals, Inc. We examined the efficacy of two ActRIIB ligand‐trapping agents in altering the morphology of a severely dystrophic muscle in the mdx mouse. The results indicate that severely dystrophic muscle exhibits monotonic postnatal increases in myonuclei and fiber density that are not associated with alterations in the distribution of fiber cross‐sectional areas. Long‐term treatment with an ActRIIB ligand‐trapping agent substantially enhanced this postnatal hyperplasia and reduced centronucleation. These results demonstrating a continual postnatal proliferation and fusion of satellite cells and a response to myostatin blockade characteristic of developing prenatal muscle suggest that the lack of dystrophin directly results in unrestrained postnatal satellite cell activation that is not necessarily dependent upon prior fiber degeneration.

  February 16, 2017   doi: 10.1002/jcp.25694   open full text
• MicroRNA‐125a‐5p Is a Downstream Effector of Sorafenib in Its Antiproliferative Activity Toward Human Hepatocellular Carcinoma Cells.
  Nicoletta Potenza, Nicola Mosca, Silvia Zappavigna, Filomena Castiello, Marta Panella, Carmela Ferri, Daniela Vanacore, Antonio Giordano, Paola Stiuso, Michele Caraglia, Aniello Russo.
  Journal of Cellular Physiology. February 16, 2017
  Sorafenib is an antitumor drug for treatment of advanced hepatocellular carcinoma (HCC). It acts as a multikinase inhibitor suppressing cell proliferation and angiogenesis. Human microRNA‐125a‐5p (miR‐125a) is endowed with similar activities and is frequently downregulated in HCC. Looking for a potential microRNA‐based mechanism of action of the drug, we found that sorafenib increases cellular expression of miR‐125a in cultured HuH‐7 and HepG2 HCC cells. Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin‐7, a NAD(+)‐dependent deacetylase, and p21/p27‐dependent cell cycle arrest in G1. Later, recruitment of miR‐125a in the antiproliferative activity of sorafenib was inquired by modulating its expression in combination with the drug treatment. This analysis showed that intracellular delivery of miR‐125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR‐125a inhibitor could counteract it. Finally, evaluation of other oncogenic targets of miR‐125a revealed its ability to interfere with the expression of matrix metalloproteinase‐11, Zbtb7a proto‐oncogene, and c‐Raf, possibly contributing to the antiproliferative activity of the drug. J. Cell. Physiol. 232: 1907–1913, 2017. © 2016 Wiley Periodicals, Inc. Upregulation of the microRNA inhibited cell proliferation by suppression of sirtuin‐7, a NAD(+)‐dependent deacetylase, and p21/p27‐dependent cell cycle arrest in G1. Intracellular delivery of miR‐125a had no additive effect on the antiproliferative activity of sorafenib, whereas a miR‐125a inhibitor could counteract it. miR‐125a interfered with the expression of matrix metalloproteinase‐11, Zbtb7a proto‐oncogene, and c‐Raf, possibly contributing to the antiproliferative activity of the drug.

  February 16, 2017   doi: 10.1002/jcp.25744   open full text
• Knockdown of REV3 synergizes with ATR inhibition to promote apoptosis induced by cisplatin in lung cancer cells.
  He‐Guo Jiang, Ping Chen, Jin‐Yu Su, Ming Wu, Hai Qian, Yi Wang, Jian Li.
  Journal of Cellular Physiology. February 13, 2017
  It has been demonstrated that REV3, the catalytic subunit of the translesion synthesis (TLS) polymerase ζ, play an important role in DNA damage response (DDR) induced by cisplatin, and Ataxia‐telangietasia mutated and Rad‐3‐related (ATR) knase is a central player in activating cell cycle checkpoint, stabilizing replication forks, regulating DDR, and promoting repair of DNA damage caused by cisplatin. Cancer cells deficient in either one of REV3 and ATR are more sensitive to cisplatin. However, whether co‐inhibition of REV3 and ATR can further increase sensitivity of non‐small cell lung cancer (NSCLC) cells to cisplatin is not clear. In this study, we show that REV3 knockdown combined with ATR inhibition further enhance cytotoxicity of cisplatin in NSCLC cells, including cisplatin‐sensitive and ‐resistant cell lines, compared to individual knockdown of REV3 or ATR, which are accompanied by markedly caspase‐dependent apoptosis response, pronounced DNA damage accumulation and severe impediment of interstrand crosslink (ICL), and double strand break (DSB) repair. Our results suggest that REV3 knockdown synergize strongly with ATR inhibition to significantly increase sensitivity of cisplatin in NSCLC cells by inhibiting ICL and DSB repair. Thus simultaneously targeting REV3 and ATR may represent one approach to overcome cisplatin resistance and improve chemotherapeutic efficacy in NSCLC treatment. Herein, we showed that depletion of REV3 combined with ATR inhibition significantly potentiates the cytotoxicity of cisplatin to non‐small cell lung cancer (NSCLC) cells, which is concomitant with pronounced DNA damage accumulation and severe impediment of interstrand crosslink (ICL) and double strand break (DSB) repair, compared to individual depletion of REV3 or ATR. Our data demonstrate that REV3 depletion can synergize with ATR inhibition to increase suppression effect of cisplatin on NSCLC cells through inhibition of ICL and DSB repair.

  February 13, 2017   doi: 10.1002/jcp.25792   open full text
• M2‐like macrophages induce colon cancer cell invasion via matrix metalloproteinases.
  Katyayni Vinnakota, Yuan Zhang, Benson Chellakkan Selvanesan, Geriolda Topi, Tavga Salim, Janna Sand‐Dejmek, Gunilla Jönsson, Anita Sjölander.
  Journal of Cellular Physiology. February 13, 2017
  The inflammatory milieu plays an important role in colon cancer development and progression. Previously, we have shown that tumor‐associated macrophages (TAMs), an important component of the tumor microenvironment, are enriched in tumors compared with normal tissue and confer a poorer prognosis. In the present study, we found that matrix metallopeptidase‐9 (MMP‐9), which degrades extracellular matrix proteins, was increased in biopsies from colon cancer patients and in mouse xenografts with SW480 cell‐derived tumors. SW480 colon cancer cells exposed to M2‐like macrophage‐conditioned medium (M2‐medium) exhibited increased MMP‐9 mRNA, protein expression and gelatinase activity. A similar effect was obtained by the addition of tumor necrosis factor‐α (TNFα) and leukotriene D4 (LTD4). MMP‐9 expression and activity were reduced by a TNFα blocking antibody adalimumab and a cysteinyl leukotriene receptor 1 (CysLTR1, the receptor for LTD4) antagonist montelukast. M2‐medium also induced changes in the epithelial–mesenchymal transition (EMT) markers E‐cadherin, β‐catenin, vimentin, and snail in SW480 cells. We also found that both M2‐medium and TNFα and LTD4 induced stabilization/nuclear translocation of β‐catenin. Furthermore, we also observed an elongated phenotype that may indicate increased invasiveness, as confirmed in a collagen I invasion assay. M2‐medium increased the invasive ability, and a similar effect was also obtained by the addition of TNFα and LTD4. The specific MMP inhibitor I or adalimumab and montelukast reduced the number of invasive cells. In conclusion, our findings show that M2‐medium enriched in TNFα and LTD4 promote colon cancer cell invasion via MMP‐9 expression and activation and the induction of EMT. Our findings show that M2‐medium enriched in TNFalfa and LTD4 promote colon cancer cell invasion via MMP‐9 expression and activation and the induction of EMT in SW480 colon cancer cells.

  February 13, 2017   doi: 10.1002/jcp.25808   open full text
• Lipid profiling of parkin‐mutant human skin fibroblasts.
  Simona Lobasso, Paola Tanzarella, Daniele Vergara, Michele Maffia, Tiziana Cocco, Angela Corcelli.
  Journal of Cellular Physiology. February 10, 2017
  Parkin mutations are a major cause of early‐onset Parkinson's disease (PD). The impairment of protein quality control system together with defects in mitochondria and autophagy process are consequences of the lack of parkin, which leads to neurodegeneration. Little is known about the role of lipids in these alterations of cell functions. In the present study, parkin‐mutant human skin primary fibroblasts have been considered as cellular model of PD to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid compositions were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined matrix‐assisted laser desorption/ionization‐time‐of‐flight mass spectrometry (MALDI‐TOF/MS) and thin‐layer chromatography (TLC). In parallel, we have performed direct MALDI‐TOF/MS lipid analyses of intact fibroblasts by skipping lipid extraction steps. Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin‐mutant fibroblasts. The detected higher level of gangliosides, phosphatidylinositol, and phosphatidylserine could be linked to dysfunction of autophagy and mitochondrial turnover; in addition, the lysophosphatidylcholine increase could represent the marker of neuroinflammatory state, a well‐known component of PD. In the present study, parkin‐mutant human skin primary fibroblasts have been considered as cellular model of Parkinson's disease (PD) in order to investigate on possible lipid alterations associated with the lack of parkin protein. Dermal fibroblasts were obtained from two unrelated PD patients with different parkin mutations and their lipid compositions were compared with that of two control fibroblasts. The lipid extracts of fibroblasts have been analyzed by combined MALDI‐TOF/MS and TLC. In parallel, we have performed direct MALDI‐TOF/MS lipid analyses of intact fibroblasts. Results show that the proportions of some phospholipids and glycosphingolipids were altered in the lipid profiles of parkin‐mutant fibroblasts.

  February 10, 2017   doi: 10.1002/jcp.25815   open full text
• Mesenchymal stem cells improves survival in LPS‐induced acute lung injury acting through inhibition of NETs formation.
  Leonardo Pedrazza, Aline Andrea Cunha, Carolina Luft, Nailê Karine Nunes, Felipe Schimitz, Rodrigo Benedetti Gassen, Ricardo Vaz Breda, Marcio Vinícius Fagundes Donadio, Angela Terezinha de Souza Wyse, Paulo Marcio Condessa Pitrez, Jose Luis Rosa, Jarbas Rodrigues de Oliveira.
  Journal of Cellular Physiology. February 09, 2017
  Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are syndromes of acute hypoxemic respiratory failure resulting from a variety of direct and indirect injuries to the gas exchange parenchyma of the lungs. During the ALI, we have an increase release of proinflammatory cytokines and high reactive oxygen species (ROS) formation. These factors are responsible for the release and activation of neutrophil‐derived proteases and the formation of neutrophil extracellular traps (NETs). The excessive increase in the release of NETs cause damage to lung tissue. Recent studies have studies involving the administration of mesenchymal stem cells (MSCs) for the treatment of experimental ALI has shown promising results. In this way, the objective of our study is to evaluate the ability of MSCs, in a lipopolysaccharide (LPS)‐induced ALI model, to reduce inflammation, oxidative damage, and consequently decrease the release of NETs. Mice were submitted lung injury induced by intratracheal instillation of LPS and subsequently treated or not with MSCs. Treatment with MSCs was able to modulate pulmonary inflammation, decrease oxidative damage, and reduce the release of NETs. These benefits from treatment are evident when we observe a significant increase in the survival curve in the treated animals. Our results demonstrate that MSCs treatment is effective for the treatment of ALI. For the first time, it is described that MSCs can reduce the formation of NETs and an experimental model of ALI. This finding is directly related to these cells modulate the inflammatory response and oxidative damage in the course of the pathology. Treatment with MSCs was able to modulate pulmonary inflammation, decrease oxidative damage, and reduce the release of NETs. For the first time, it is described that MSCs can reduce the formation of NETs and an experimental model of ALI.

  February 09, 2017   doi: 10.1002/jcp.25816   open full text
• Impact of lysosomal storage disorders on biology of mesenchymal stem cells: Evidences from in vitro silencing of glucocerebrosidase (GBA) and alpha‐galactosidase A (GLA) enzymes.
  Tiziana Squillaro, Ivana Antonucci, Nicola Alessio, Anna Esposito, Marilena Cipollaro, Mariarosa Anna Beatrice Melone, Gianfranco Peluso, Liborio Stuppia, Umberto Galderisi.
  Journal of Cellular Physiology. February 07, 2017
  Lysosomal storage disorders (LDS) comprise a group of rare multisystemic diseases resulting from inherited gene mutations that impair lysosomal homeostasis. The most common LSDs, Gaucher disease (GD), and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha‐galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected. Among stem cells, mesenchymal stem cells (MSCs) are a commonly investigated population given their role in hematopoiesis and the homeostatic maintenance of many organs and tissues. Since the impairment of MSC functions could pose profound consequences on body physiology, we evaluated whether GBA and GLA silencing could affect the biology of MSCs isolated from bone marrow and amniotic fluid. Those cell populations were chosen given the former's key role in organ physiology and the latter's intriguing potential as an alternative stem cell model for human genetic disease. Our results revealed that GBA and GLA deficiencies prompted cell cycle arrest along with the impairment of autophagic flux and an increase of apoptotic and senescent cell percentages. Moreover, an increase in ataxia–telangiectasia‐mutated staining 1 hr after oxidative stress induction and a return to basal level at 48 hr, along with persistent gamma‐H2AX staining, indicated that MSCs properly activated DNA repair signaling, though some damages remained unrepaired. Our data therefore suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity. Gaucher disease (GD) and Fabry disease (FD) are caused by deficiencies in the lysosomal glucocerebrosidase (GBA) and alpha‐galactosidase A (GLA) enzymes, respectively. Given the systemic nature of enzyme deficiency, we hypothesized that the stem cell compartment of GD and FD patients might be also affected; thus, we evaluated whether GBA and GLA silencing could affect the biology of mesenchymal stem cells (MSCs) isolated from bone marrow and amniotic fluid. Our data suggest that MSCs with reduced GBA or GLA activity are prone to apoptosis and senescence due to impaired autophagy and DNA repair capacity.

  February 07, 2017   doi: 10.1002/jcp.25807   open full text
• LSP1, a responsive protein from Meyerozyma guilliermondii, elicits defence response and improves glycyrrhizic acid biosynthesis in Glycyrrhiza uralensis Fisch adventitious roots.
  Juan Wang, Jianli Li, Jing Li, Jinxin Li, Shujie Liu, Wenyuan Gao.
  Journal of Cellular Physiology. February 07, 2017
  This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the Meyerozyma guilliermondii since the excellent induction. The contents of total flavonoids (3.46 mg · g−1), glycyrrhizic acid (0.41 mg · g−1), glycyrrhetinic acid (0.41 mg · g−1), and polysaccharide (94.49 mg · g−1) in adventitious root peaked at LSP1 group, which were 1.6, 3.4, 2.4, 2.0‐fold that of control, respectively. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen‐activated protein kinases and extremely up‐regulated the expression of defense‐related genes and functional genes involved in glycyrrhizic acid biosynthesis. This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the M. guilliermondii since the excellent induction. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen‐activated protein kinases and extremely up‐regulated the expression of defense‐related genes and functional genes involved in glycyrrhizic acid biosynthesis.

  February 07, 2017   doi: 10.1002/jcp.25811   open full text
• Involvement of CX3CL1 in the Migration of Osteoclast Precursors Across Osteoblast Layer Stimulated by Interleukin‐1ß.
  Tsuyoshi Matsuura, Shizuko Ichinose, Masako Akiyama, Yuki Kasahara, Noriko Tachikawa, Ken‐ichi Nakahama.
  Journal of Cellular Physiology. February 03, 2017
  The trigger for bone remodeling is bone resorption by osteoclasts. Osteoclast differentiation only occurs on the old bone, which needs to be repaired under physiological conditions. However, uncontrolled bone resorption is often observed in pro‐inflammatory bone diseases, such as rheumatoid arthritis. Mature osteoclasts are multinuclear cells that differentiate from monocyte/macrophage lineage cells by cell fusion. Although Osteoclast precursors should migrate across osteoblast layer to reach bone matrix before maturation, the underlying mechanisms have not yet been elucidated in detail. We herein found that osteoclast precursors utilize two routes to migrate across osteoblast layer by confocal‐ and electro‐microscopic observations. The osteoclast supporting activity of osteoblasts inversely correlated with osteoblast density and was positively related to the number of osteoclast precursors under the osteoblast layer. Osteoclast differentiation was induced by IL‐1ß, but not by PGE2 in high‐density osteoblasts. Osteoblasts and osteoclast precursors expressed CX3CL1 and CX3CR1, respectively, and the expression of CX3CL1 increased in response to interleukin‐1ß. An anti‐CX3CL1‐neutralizing antibody inhibited the migration of osteoclast precursors and osteoclast differentiation. These results strongly suggest the involvement of CX3CL1 in the migration of osteoclast precursors and osteoclastogenesis, and will contribute to the development of new therapies for bone diseases. J. Cell. Physiol. 232: 1739–1745, 2017. © 2016 Wiley Periodicals, Inc. Transcellular and paracellular migration of osteoclast precursors across osteoblast layer were observed. CX3CL1 was involved in the migration of osteoclast precursors and osteoclastogenesis.

  February 03, 2017   doi: 10.1002/jcp.25577   open full text
• DNA Methylation Profiling in Chondrocyte Dedifferentiation In Vitro.
  Li Duan, Yujie Liang, Bin Ma, Daming Wang, Wei Liu, Jianghong Huang, Jianyi Xiong, Liangquan Peng, Jielin Chen, Weimin Zhu, Daping Wang.
  Journal of Cellular Physiology. January 31, 2017
  DNA methylation has emerged as a crucial regulator of chondrocyte dedifferentiation, which severely compromises the outcome of autologous chondrocyte implantation (ACI) treatment for cartilage defects. However, the full‐scale DNA methylation profiling in chondrocyte dedifferentiation remains to be determined. Here, we performed a genome‐wide DNA methylation profiling of dedifferentiated chondrocytes in monolayer culture and chondrocytes treated with DNA methylation inhibitor 5‐azacytidine (5‐AzaC). This research revealed that the general methylation level of CpG was increased while the COL‐1A1 promoter methylation level was decreased during the chondrocyte dedifferentiation. 5‐AzaC could reduce general methylation levels and reverse the chondrocyte dedifferentiation. Surprisingly, the DNA methylation level of COL‐1A1 promoter was increased after 5‐AzaC treatment. The COL‐1A1 expression level was increased while that of SOX‐9 was decreased during the chondrocyte dedifferentiation. 5‐AzaC treatment up‐regulated the SOX‐9 expression while down‐regulated the COL‐1A1 promoter activity and gene expression. Taken together, these results suggested that differential regulation of the DNA methylation level of cartilage‐specific genes might contribute to the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation. J. Cell. Physiol. 232: 1708–1716, 2017. © 2016 Wiley Periodicals, Inc. Differential regulation of the DNA methylation level of cartilage‐specific genes might contribute the chondrocyte dedifferentiation. Thus, the epigenetic manipulation of these genes could be a potential strategy to counteract the chondrocyte dedifferentiation accompanying in vitro propagation.

  January 31, 2017   doi: 10.1002/jcp.25486   open full text
• Cytoskeleton Aberrations in Alkaptonuric Chondrocytes.
  Michela Geminiani, Silvia Gambassi, Lia Millucci, Pietro Lupetti, Giulia Collodel, Lucia Mazzi, Bruno Frediani, Daniela Braconi, Barbara Marzocchi, Marcella Laschi, Giulia Bernardini, Annalisa Santucci.
  Journal of Cellular Physiology. January 31, 2017
  Alkaptonuria (AKU) is an ultra‐rare autosomal genetic disorder caused by a defect in the activity of the enzyme homogentisate 1,2‐dioxygenase (HGD) that leads to the accumulation of homogentisic acid (HGA) and its oxidized product, benzoquinone acetic acid (BQA), in the connective tissues causing a pigmentation called “ochronosis.” The consequent progressive formation of ochronotic aggregates generate a severe condition of oxidative stress and inflammation in all the affected areas. Experimental evidences have also proved the presence of serum amyloid A (SAA) in several AKU tissues and it allowed classifying AKU as a secondary amyloidosis. Although AKU is a multisystemic disease, the most affected system is the osteoarticular one and articular cartilage is the most damaged tissue. In this work, we have analyzed for the first time the cytoskeleton of AKU chondrocytes by means of immunofluorescence staining. We have shown the presence of SAA within AKU chondrocytes and finally we have demonstrated the co‐localization of SAA with three cytoskeletal proteins: actin, vimentin, and β‐tubulin. Furthermore, in order to observe the ultrastructural features of AKU chondrocytes we have performed TEM analysis, focusing on the Golgi apparatus structure and, to demonstrate that pigmented areas in AKU cartilage are correspondent to areas of oxidation, 4‐HNE presence has been evaluated by means of immunofluorescence. J. Cell. Physiol. 232: 1728–1738, 2017. © 2016 Wiley Periodicals, Inc. The cytoskeleton of alkaptonuric chondrocytes is severely alterated. Serum amyloid A is demonstrated to be present within alkaptonuric chondrocytes and to co‐localize with the three main cytoskeletal proteins: actin, vimentin, and β‐tubulin. These findings lead to hypothesize a role of SAA in cytoskeleton alterations of alkaptonuric chondrocytes.

  January 31, 2017   doi: 10.1002/jcp.25500   open full text
• Exosomes: Nanoparticulate tools for RNA interference and drug delivery.
  Fahimeh Shahabipour, Nastaran Barati, Thomas P. Johnston, Giuseppe Derosa, Pamela Maffioli, Amirhossein Sahebkar.
  Journal of Cellular Physiology. January 31, 2017
  Exosomes are naturally occurring extracellular vesicles released by most mammalian cells in all body fluids. Exosomes are known as key mediators in cell‐cell communication and facilitate the transfer of genetic and biochemical information between distant cells. Structurally, exosomes are composed of lipids, proteins, and also several types of RNAs which enable these vesicles to serve as important disease biomarkers. Moreover, exosomes have emerged as novel drug and gene delivery tools owing to their multiple advantages over conventional delivery systems. Recently, increasing attention has been focused on exosomes for the delivery of drugs, including therapeutic recombinant proteins, to various target tissues. Exosomes are also promising vehicles for the delivery of microRNAs and small interfering RNAs, which is usually hampered by rapid degradation of these RNAs, as well as inefficient tissue specificity of currently available delivery strategies. This review highlights the most recent accomplishments and trends in the use of exosomes for the delivery of drugs and therapeutic RNA molecules. Exosomes are naturally occurring extracellular vesicles released by most mammalian cells in all body fluids. Recently, increasing attention has been focused on exosomes for the delivery of drugs, including therapeutic recombinant proteins, to various target tissues. This review highlights the most recent accomplishments and trends in the use of exosomes for the delivery of drugs and therapeutic RNA molecules.

  January 31, 2017   doi: 10.1002/jcp.25766   open full text
• Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway.
  Maria M. Barreca, Walter Spinello, Vincenzo Cavalieri, Giuseppina Turturici, Gabriella Sconzo, Punit Kaur, Rosaria Tinnirello, Alexzander A. A. Asea, Fabiana Geraci.
  Journal of Cellular Physiology. January 31, 2017
  Mouse mesoangioblasts are vessel‐associated progenitor stem cells endowed with the ability of multipotent mesoderm differentiation. Therefore, they represent a promising tool in the regeneration of injured tissues. Several studies have demonstrated that homing of mesoangioblasts into blood and injured tissues are mainly controlled by cytokines/chemokines and other inflammatory factors. However, little is known about the molecular mechanisms regulating their ability to traverse the extracellular matrix (ECM). Here, we demonstrate that membrane vesicles released by mesoangioblasts contain Hsp70, and that the released Hsp70 is able to interact by an autocrine mechanism with Toll‐like receptor 4 (TLR4) and CD91 to stimulate migration. We further demonstrate that Hsp70 has a positive role in regulating matrix metalloproteinase 2 (MMP2) and MMP9 expression and that MMP2 has a more pronounced effect on cell migration, as compared to MMP9. In addition, the analysis of the intracellular pathways implicated in Hsp70 regulated signal transduction showed the involvement of both PI3K/AKT and NF‐κB. Taken together, our findings present a paradigm shift in our understanding of the molecular mechanisms that regulate mesoangioblast stem cells ability to traverse the extracellular matrix (ECM). J. Cell. Physiol. 232: 1845–1861, 2017. © 2016 Wiley Periodicals, Inc. Extracellular Hsp70 released through extracellular vesicles regulates mesoangioblast migration via the activation of NF‐κB and PI3K/AKT pathways.

  January 31, 2017   doi: 10.1002/jcp.25722   open full text
• HDAC1, HDAC4, and HDAC9 Bind to PC3/Tis21/Btg2 and Are Required for Its Inhibition of Cell Cycle Progression and Cyclin D1 Expression.
  Laura Micheli, Giorgio D'Andrea, Luca Leonardi, Felice Tirone.
  Journal of Cellular Physiology. January 27, 2017
  PC3/Tis21 is a transcriptional cofactor that inhibits proliferation in several cell types, including neural progenitors. Here, we report that PC3/Tis21 associates with HDAC1, HDAC4, and HDAC9 in vivo, in fibroblast cells. Furthermore, when HDAC1, HDAC4, or HDAC9 are silenced in fibroblasts or in a line of cerebellar progenitor cells, the ability of PC3/Tis21 to inhibit proliferation is significantly reduced. Overexpression of HDAC1, HDAC4, or HDAC9 in fibroblasts and in cerebellar precursor cells synergizes with PC3/Tis21 in inhibiting the expression of cyclin D1, a cyclin selectively inhibited by PC3/Tis21. Conversely, the depletion of HDAC1 or HDAC4 (but not HDAC9) in fibroblasts and in cerebellar precursor cells significantly impairs the ability of PC3/Tis21 to inhibit cyclin D1 expression. An analysis of HDAC4 deletion mutants shows that both the amino‐terminal moiety and the catalytic domain of HDAC4 associate to PC3/Tis21, but neither alone is sufficient to potentiate the inhibition of cyclin D1 by PC3/Tis21. As a whole, our findings indicate that PC3/Tis21 inhibits cell proliferation in a way dependent on the presence of HDACs, in fibroblasts as well as in neural cells. Considering that several reports have demonstrated that HDACs can act as transcriptional corepressors on the cyclin D1 promoter, our data suggest that the association of PC3/Tis21 to HDACs is functional to recruit them to target genes, such as cyclin D1, for repression of their expression. J. Cell. Physiol. 232: 1696–1707, 2017. © 2016 Wiley Periodicals, Inc. By silencing HDAC1, HDAC4, and HDAC9 in fibroblast as well as in neural cells, we reveal that HDAC1, HDAC4, and HDAC9 are each required for the negative control of the cell cycle exerted by PC3/Tis21. Such effect is likely dependent on the ability of HDAC1, HDAC4, and HDAC9 to associate with PC3/Tis21, that we demonstrate here. Moreover, we find that HDAC1 and HDAC4—but not HDAC9—are required for the inhibition exerted by PC3/Tis21 of cyclin D1 expression.

  January 27, 2017   doi: 10.1002/jcp.25467   open full text
• State of the Art in MicroRNA as Diagnostic and Therapeutic Biomarkers in Chronic Lymphocytic Leukemia.
  Hamed Mirzaei, Sima Fathullahzadeh, Razieh Khanmohammadi, Mansoreh Darijani, Fatemeh Momeni, Aria Masoudifar, Mohammad Goodarzi, Omid Mardanshah, Jan Stanveng, Mahmoud Reza Jaafari, Hamid Reza Mirzaei.
  Journal of Cellular Physiology. January 13, 2017
  Early diagnostic is one of the most important steps in cancer therapy which helps to design and choose a better therapeutic approach. The finding of biomarkers in various levels including genomics, transcriptomics and proteomics levels could provide better treatment for various cancers such as chronic lymphocytic leukemia (CLL). The CLL is the one of main lymphoid malignancies which is specified by aggregation of mature B lymphocytes. Among different biomarkers (e.g. CD38, chromosomes abnormalities, ZAP‐70, TP53 and miRNA), microRNAs (miRNAs) have appeared as new diagnostic and therapeutic biomarkers in patients with the CLL disease. Multiple lines of evidence indicated that deregulation of miRNAs could be associated with pathological events which are present in the CLL. These molecules have an effect on a variety of targets such as Bcl2, c‐fos, c‐Myc, TP53, TCL1 and STAT3 which play critical roles in the CLL pathogenesis. It has been shown that expression of miRNAs could lead to the activation of B cells and BCR. Moreover, exosomes containing miRNAs are one of the other molecules which could contribute to BCR stimulation and progression of CLL cells. Hence, miRNAs and exosomes released from CLL cells could be used as potential diagnostic and therapeutic biomarkers for CLL. This critical review focuses on a very important aspect of CLL based on biomarker discovery covers the pros and cons of using miRNAs as important diagnostics and therapeutics biomarkers for this deadly disease. This article is protected by copyright. All rights reserved

  January 13, 2017   doi: 10.1002/jcp.25799   open full text
• Gating Modulation of the Tumor‐Related Kv10.1 Channel by Mibefradil.
  Froylán Gómez‐Lagunas, Elisa Carrillo, Luis A. Pardo, Walter Stühmer.
  Journal of Cellular Physiology. January 11, 2017
  Several reports credit mibefradil with tumor suppressing properties arising from its known inhibition of Ca2+ currents. Given that mibefradil (Mb) is also known to inhibit K+ channels, we decided to study the interaction between this organic compound and the tumor‐related Kv10.1 channel. Here we report that Mb modulates the gating of Kv10.1. Mb induces an apparent inactivation from both open and early closed states where the channels dwell at hyperpolarized potentials. Additionally, Mb accelerates the kinetics of current activation, in a manner that depends on initial conditions. Our observations suggest that Mb binds to the voltage sensor domain of Kv10.1 channels, thereby modifying the gating of the channels in a way that in some, but not all, aspects opposes to the gating effects exerted by divalent cations. J. Cell. Physiol. 232: 2019–2032, 2017. © 2016 Wiley Periodicals, Inc. Several reports credit mibefradil, a T‐type calcium channel inhibitor, with tumor suppressing properties. Given that mibefradil also inhibits some potassium channels, we studied the interaction of this compound with the tumor related Kv10.1 channel. We found that mibefradil induces an apparent inactivation both from the open and early closed states, and accelerates current activation in a manner that depends on the holding potential. We hypothesize that mibefradil binds to the voltage‐sensor module altering the gating of the channels.

  January 11, 2017   doi: 10.1002/jcp.25448   open full text
• HOTAIR Role in Melanoma Progression and its Identification in the Blood of Patients with Advanced Disease.
  Monica Cantile, Giosuè Scognamiglio, Laura Marra, Gabriella Aquino, Chiara Botti, Maria Rosaria Falcone, Maria Gabriella Malzone, Giuseppina Liguori, Maurizio Di Bonito, Renato Franco, Paolo Antonio Ascierto, Gerardo Botti.
  Journal of Cellular Physiology. January 09, 2017
  Objectives The molecular mechanisms responsible for the metastatic progression of melanoma have not been fully defined yet. We have recently shown that an important role in this process is certainly played by HOX genes, whose regulation is under control of particular non‐coding RNAs, some of which are present within the HOX locus. HOTAIR is the most studied among them, whose aberrant expression is associated with the metastatic progression of many malignancies. The aim of this study was to verify the role played by HOTAIR in metastatic progression of melanoma and to evaluate the circulating levels of HOTAIR in the blood of patients with metastatic melanoma. Methods A series of melanocytic lesions were selected to evaluate the potential changes in the expression of HOTAIR during the evolution of the disease through in situ and molecular approaches. Results None of the benign melanocytic lesions showed the presence of HOTAIR. The staining of HOTAIR resulted very weak in the primary pT1 lesions, while it was very strong in all pairs of primary tissues and corresponding metastases. Surprisingly, we found the presence of HOTAIR in some intratumoral lymphocytes, while this positivity decreased in lymphocyte component further away from the tumor. HOTAIR was also detected in the serum of selected metastatic patients. Conclusion These data allowed us to speculate on the fundamental role played by HOTAIR in tumor evolution of melanoma. Its presence in intratumoral lymphocytes might suggest that its involvement in the modulation of tumor microenvironment and the detection in the serum could be used in the management of melanoma patients. This article is protected by copyright. All rights reserved

  January 09, 2017   doi: 10.1002/jcp.25789   open full text
• Biological Roles of Glial Fibrillary Acidic Protein as a Biomarker in Cartilage Regenerative Medicine.
  Sanshiro Kanazawa, Satoru Nishizawa, Tsuyoshi Takato, Kazuto Hoshi.
  Journal of Cellular Physiology. January 07, 2017
  Glial fibrillary acidic protein (GFAP) is an intermediate filament that is expressed in specifically expressed auricular chondrocytes, which are good cell sources of cartilage regenerative medicine. Although our group uses GFAP as a biomarker of matrix production in the cultured auricular chondrocytes, the biological roles of GFAP in auricular chondrocytes has remained unknown. In this study, we demonstrated the biological functions of GFAP in the human and mouse derived auricles to clarify the significance and role with the chondrocytes of GFAP in order to provide useful information for reliable and safe regenerative medicine. We examined the cell responses to stretch stress for these chondrocytes and completed a nuclear morphological analysis. Based on these results, GFAP seems to support the resistance to severe mechanical stress in the tissue which physiologically suffers from a stretch overload, and plays pivotal roles in the conservation of cell structures and functions through the maintenance of nuclear morphology. This article is protected by copyright. All rights reserved

  January 07, 2017   doi: 10.1002/jcp.25771   open full text
• Optimized Method for Isolating Highly Purified and Functional Porcine Aortic Endothelial and Smooth Muscle Cells.
  Farideh Beigi, Mitalben Patel, Marco A. Morales‐Garza, Caitlin Winebrenner, Andrea S. Gobin, Eric Chau, Luiz C. Sampaio, Doris A. Taylor.
  Journal of Cellular Physiology. January 06, 2017
  Background Numerous protocols exist for isolating aortic endothelial and smooth muscle cells from small animals. However, establishing a protocol for isolating pure cell populations from large animal vessels that are more elastic has been challenging. We developed a simple sequential enzymatic approach to isolate highly purified populations of porcine aortic endothelial and smooth muscle cells. Methods The lumen of a porcine aorta was filled with 25 U/mL dispase solution and incubated at 37°C to dissociate the endothelial cells. The smooth muscle cells were isolated by mincing the tunica media of the treated aorta and incubating the pieces in 0.2% and then 0.1% collagenase type I solution. Results The isolated endothelial cells stained positive for von Willebrand factor, and 97.2% of them expressed CD31. Early‐ and late‐passage endothelial cells had a population doubling time of 38 hours and maintained a capacity to take up DiI‐Ac‐LDL and form tubes in Matrigel®. The isolated smooth muscle cells stained highly positive for alpha‐smooth muscle actin, and an impurities assessment showed that only 1.8% were endothelial cells. Population doubling time for the smooth muscle cells was ∼70 hours at passages 3 and 7; and the cells positively responded to endothelin‐1, as shown by a 66% increase in the intracellular calcium level. Conclusions This simple protocol allows for the isolation of highly pure populations of endothelial and smooth muscle cells from porcine aorta that can survive continued passage in culture without losing functionality or becoming overgrown by fibroblasts. This article is protected by copyright. All rights reserved

  January 06, 2017   doi: 10.1002/jcp.25764   open full text
• SIRT1‐SIRT3 Axis Regulates Cellular Response to Oxidative Stress and Etoposide.
  Ilaria Carnevale, Laura Pellegrini, Patrizia D'Aquila, Serena Saladini, Emanuela Lococo, Lucia Polletta, Enza Vernucci, Eleonora Foglio, Stefano Coppola, Luigi Sansone, Giuseppe Passarino, Dina Bellizzi, Matteo A. Russo, Massimo Fini, Marco Tafani.
  Journal of Cellular Physiology. January 06, 2017
  Sirtuins are conserved NAD+‐dependent deacylases. SIRT1 is a nuclear and cytoplasmic sirtuin involved in the control of histones a transcription factors function. SIRT3 is a mitochondrial protein, which regulates mitochondrial function. Although, both SIRT1 and SIRT3 have been implicated in resistance to cellular stress, the link between these two sirtuins has not been studied so far. Here we aimed to unravel: i) the role of SIRT1‐SIRT3 axis for cellular response to oxidative stress and DNA damage; ii) how mammalian cells modulate such SIRT1‐SIRT3 axis and which mechanisms are involved. Therefore, we analyzed the response to different stress stimuli in WT or SIRT1‐silenced cell lines. Our results demonstrate that SIRT1‐silenced cells are more resistant to H2O2 and etoposide treatment showing decreased ROS accumulation, γ‐H2AX phosphorylation, caspase‐3 activation and PARP cleavage. Interestingly, we observed that SIRT1‐silenced cells show an increased SIRT3 expression. To explore such a connection, we carried out luciferase assays on SIRT3 promoter demonstrating that SIRT1‐silencing increases SIRT3 promoter activity and that such an effect depends on the presence of SP1 and ZF5 recognition sequences on SIRT3 promoter. Afterwards, we performed co‐immunoprecipitation assays demonstrating that SIRT1 binds and deacetylates the transcription inhibitor ZF5 and that there is a decreased interaction between SP1 and ZF5 in SIRT1‐silenced cells. Therefore, we speculate that acetylated ZF5 cannot bind and sequester SP1 that is free, then, to increase SIRT3 transcription. In conclusion, we demonstrate that cells with low SIRT1 levels can maintain their resistance and survival by increasing SIRT3 expression. J. Cell. Physiol. 232: 1835–1844, 2017. © 2016 Wiley Periodicals, Inc. Our article demonstrate that cells with low SIRT1 levels can maintain their resistance and survival to exogenous stresses by increasing SIRT3 expression.

  January 06, 2017   doi: 10.1002/jcp.25711   open full text
• Calprotectin Induces IL‐6 and MCP‐1 Production via Toll‐Like Receptor 4 Signaling in Human Gingival Fibroblasts.
  Yasufumi Nishikawa, Yukari Kajiura, Jung Hwan Lew, Jun‐ichi Kido, Toshihiko Nagata, Koji Naruishi.
  Journal of Cellular Physiology. January 06, 2017
  Calprotectin, a heterodimer of S100A8 and S100A9 molecules, is associated with inflammatory diseases such as inflammatory bowel disease. We have reported that calprotectin levels in gingival crevicular fluids of periodontitis patients are significantly higher than in healthy subjects. However, the functions of calprotectin in pathophysiology of periodontitis are still unknown. The aim of this study is to investigate the effects of calprotectin on the productivity of inflammatory cytokines in human gingival fibroblasts (HGFs). The HGFs cell line CRL‐2014® (ATCC) were cultured, and total RNAs were collected to examine the expression of TLR2/4 and RAGE mRNA using RT‐PCR. After the cells were treated with S100A8, S100A9, and calprotectin, supernatants were collected and the levels of IL‐6 and MCP‐1 were measured using ELISA methods. To examine the intracellular signals involved in calprotectin‐induced cytokine production, several chemical inhibitors were used. Furthermore, after the siRNA‐mediated TLR4 down‐regulated cells were treated with S100A8, S100A9, and calprotectin, the levels of IL‐6 and MCP‐1 were also measured. HGFs showed greater expression of TLR4 mRNA, but not TLR2 and RAGE mRNA compared with human oral epithelial cells. Calprotectin increased significantly the production of MCP‐1 and IL‐6 in HGFs, and the cytokine productions were significantly suppressed in the cells treated with MAPKs, NF‐κB, and TLR4 inhibitors. Furthermore, calprotectin‐mediated MCP‐1 and IL‐6 production were significantly suppressed in TLR4 down‐regulated cells. Taken together, calprotectin induces IL‐6 and MCP‐1 production in HGFs via TLR4 signaling that involves MAPK and NF‐κB, resulting in the progression of periodontitis. J. Cell. Physiol. 232: 1862–1871, 2017. © 2016 Wiley Periodicals, Inc.

  January 06, 2017   doi: 10.1002/jcp.25724   open full text
• Cyclic Tensile Strain Reduces TNF‐α Induced Expression of MMP‐13 by Condylar Temporomandibular Joint Cells.
  Hessam Tabeian, Astrid D. Bakker, Beatriz F. Betti, Frank Lobbezoo, Vincent Everts, Teun J. de Vries.
  Journal of Cellular Physiology. January 06, 2017
  To investigate whether the disproportionate degradation of mandibular condyle cartilage in arthritic juvenile temporomandibular joint (TMJ) is related to distinctive responses of TMJ‐derived cells to tumor necrosis factor‐α (TNF‐α), and whether mechanical loading affects this response. The effect of TNF‐α (0.1–10 ng/ml) was tested on juvenile porcine TMJ cells isolated from the condyle, fossa, and disc, grown in 3D agarose gels. Expression of anabolic and catabolic factors was quantified by RT‐qPCR and/or immunohistochemistry. Condylar cells were stimulated for 12 h with TNF‐α (10 ng/ml), followed by 8 h of 6% cyclic tensile strain, and gene expression of MMPs was quantified. TNF‐α (10 ng/ml) reduced the expression of the matrix proteins collagen types I and II after 6 h of incubation. Aggrecan gene expression was increased in the presence of 0.1 ng/ml TNF‐α. The fossa and disc cells responded to TNF‐α with an increased expression of the aggrecanase ADAMTS4. TNF‐α enhanced MMP‐13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect. Cells isolated from the different cartilaginous structures reacted differently to TNF‐α. Since the disc and fossa contain a very low level of proteoglycans in comparison to the condyle, the role played by ADAMTS4 in degradation of the fossa and disc might be limited. TNF‐α induced MMP‐13 expression by condylar cells might be involved in the degradation of the juvenile condyle. Since this expression was reduced by mechanical loading, functional loading with oral physiotherapy or orthodontic activators may help to reduce the catabolic effect of TNF‐α. J. Cell. Physiol. 232: 1287–1294, 2017. © 2016 Wiley Periodicals, Inc. Cells isolated from the different cartilaginous structures reacted differently to TNF‐α. TNF‐α enhanced MMP‐13 gene and protein expression only by condylar cells. Mechanical loading reduced this effect.

  January 06, 2017   doi: 10.1002/jcp.25593   open full text
• A MicroRNA Cluster miR‐23–24–27 Is Upregulated by Aldosterone in the Distal Kidney Nephron Where it Alters Sodium Transport.
  Xiaoning Liu, Robert S. Edinger, Christine A. Klemens, Yu L. Phua, Andrew J. Bodnar, William A. LaFramboise, Jacqueline Ho, Michael B. Butterworth.
  Journal of Cellular Physiology. January 05, 2017
  The epithelial sodium channel (ENaC) is expressed in the epithelial cells of the distal convoluted tubules, connecting tubules, and cortical collecting duct (CCD) in the kidney nephron. Under the regulation of the steroid hormone aldosterone, ENaC is a major determinant of sodium (Na+) and water balance. The ability of aldosterone to regulate microRNAs (miRs) in the kidney has recently been realized, but the role of miRs in Na+ regulation has not been well established. Here we demonstrate that expression of a miR cluster mmu‐miR‐23–24–27, is upregulated in the CCD by aldosterone stimulation both in vitro and in vivo. Increasing the expression of these miRs increased Na+ transport in the absence of aldosterone stimulation. Potential miR targets were evaluated and miR‐27a/b was verified to bind to the 3′‐untranslated region of intersectin‐2, a multi‐domain protein expressed in the distal kidney nephron and involved in the regulation of membrane trafficking. Expression of Itsn2 mRNA and protein was decreased after aldosterone stimulation. Depletion of Itsn2 expression, mimicking aldosterone regulation, increased ENaC‐mediated Na+ transport, while Itsn2 overexpression reduced ENaC's function. These findings reinforce a role for miRs in aldosterone regulation of Na+ transport, and implicate miR‐27 in aldosterone's action via a novel target. J. Cell. Physiol. 232: 1306–1317, 2017. © 2016 Wiley Periodicals, Inc. The steroid hormone aldosterone alters the expression of microRNAs in distal kidney nephron epithelial cells, and coordinately upregulates the miR‐23–24–27 cluster. MiR‐27 targets intersectin 2 to reduce its expression. Decreased intersectin 2 levels results in an increase in the activity of the epithelial sodium channel to augment sodium transport.

  January 05, 2017   doi: 10.1002/jcp.25599   open full text
• Early Subchondral Bone Loss at Arthritis Onset Predicted Late Arthritis Severity in a Rat Arthritis Model.
  Guillaume Courbon, Damien Cleret, Marie‐Thérèse Linossier, Laurence Vico, Hubert Marotte.
  Journal of Cellular Physiology. January 05, 2017
  Synovitis is usually observed before loss of articular function in rheumatoid arthritis (RA). In addition to the synovium and according to the “Inside–Outside” theory, bone compartment is also involved in RA pathogenesis. Then, we investigated time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant‐induced arthritis (AIA) model. Lewis female rats were longitudinally monitored from arthritis induction (day 0), with early (day 10) and late (day 17) steps. Trabecular and cortical microarchitecture parameters of four ankle bones were assessed by microcomputed tomography. Gene expression was determined at sacrifice. Arthritis occurred at day 10 in AIA rats. At this time, bone erosions were detected on four ankle bones, with cortical porosity increase (+67%) and trabecular alterations including bone volume fraction (BV/TV: −13%), and trabecular thickness decrease. Navicular bone assessment was the most reproducible and sensitive. Furthermore, strong correlations were observed between bone alterations at day 10 and arthritis severity or bone loss at day 17, including predictability of day 10 BV/TV to day 17 articular index (R2 = 0.76). Finally, gene expression at day 17 confirmed massive osteoclast activation and interestingly provided insights on strong activation of bone formation inhibitor markers at the joint level. In rat AIA, bone loss was already observed at synovitis onset and was predicted late arthritis severity. Our results reinforced the key role of subchondral bone in arthritis pathogenesis, in favour to the “Inside–Outside” theory. Mechanisms of bone loss in rat AIA involved resorption activation and formation inhibition changes. J. Cell. Physiol. 232: 1318–1325, 2017. © 2016 Wiley Periodicals, Inc. We explored time dependent articular bone loss and prediction of early bone loss to late arthritis severity on the rat adjuvant‐induced arthritis (AIA) model. At clinical synovitis onset, bone porosity and microarchitecture alterations were recorded in the ankle bones. These early modifications were predictive of arthritis outcome, underlining the importance of early bone loss investigation.

  January 05, 2017   doi: 10.1002/jcp.25601   open full text
• Calretinin Immunoreactivity in the Human Testis Throughout Fetal Life.
  Giovanna G. Altobelli, Francesca Pentimalli, Mariarosaria D'Armiento, Susan Van Noorden, Vincenzo Cimini.
  Journal of Cellular Physiology. January 05, 2017
  The main functions of the testis are sex hormone and sperm cell production. Steroidogenesis occurs in the Leydig interstitial cells and spermatogenesis in the seminiferous tubules. Male gonad morphogenesis is a finely orchestrated process, mainly coordinated by hormones, whose actions can significantly affect post‐pubertal testicular function. Calcium is a key intracellular messenger, which regulates many signal transduction pathways, and is also implicated in steroidogenesis. Calcium homeostasis and signaling rely on many calcium‐binding proteins including calretinin, of the “EF‐hand” protein family. Calretinin is a highly conserved protein mainly expressed in the nervous system but also detected in rat and human adult and fetal testis as well as in pathological conditions. Calretinin expression in the fetal testis, however, has not been thoroughly analyzed probably owing to limited availability and paucity of tissues. Here, we examined by immunocytochemistry the expression of calretinin in human fetal testis specimens, obtained from natural and therapeutic abortions, at various developmental ages. We found that calretinin‐immunoreactive Leydig cells were visible throughout the timeframe studied (14th–27th week). Immunoreactivity was also observed in Sertoli cells and in the germ cells of the immature seminiferous tubules. Overall our data indicate that calretinin expression parallels the decline in Leydig cell number, suggesting that its presence is indeed correlated to their steroidogenic activity. They also suggest that the intratubular positivity of calretinin could be linked to the ability of Sertoli cells to produce locally acting hormones contributing to the histodifferentiation of the male genital tract. J. Cell. Physiol. 232: 1872–1878, 2017. © 2016 Wiley Periodicals, Inc. Human fetal testis specimens, obtained upon natural and therapeutic abortions at various developmental ages (weeks 14–27), show calretinin expression in the Leydig interstitial cells, in the epididymis and in the immature seminiferous tubules. Calretinin expression parallels the decline in Leydig cell number. The presence of calretinin seems correlated to the Leydig cells steroidogenic activity, which is consistent with the role that calcium intracellular signaling plays in the steroidogenic process. Our findings, by adding calretinin to the list of players underlying fetal testis development, are particularly timely and prompt further assessment of calretinin role in this context both in normal and pathological conditions.

  January 05, 2017   doi: 10.1002/jcp.25727   open full text
• Fibroblast Growth Factor 21 (FGF21) Promotes Formation of Aerobic Myofibers via the FGF21‐SIRT1‐AMPK‐PGC1α Pathway.
  Xinyi Liu, Yongliang Wang, Liming Hou, Yuanzhu Xiong, Shuhong Zhao.
  Journal of Cellular Physiology. January 05, 2017
  Fibroblast growth factor 21(FGF21) is a pivotal regulator of energy metabolism, which is currently being assessed as a potential drug target for the treatment of insulin‐resistant conditions. However, the cellular mechanisms by which FGF21 affects myogenesis remain unclear. In this study, we explored the function of FGF21 in myogenesis both in vitro and in vivo. Our experiments showed for the first time that FGF21 promotes myoblast differentiation and serves as a switch of molecular transformation from anaerobic myofibers to aerobic myofibers via the FGF21‐SIRT1‐AMPK‐PGC1α axis. Furthermore, we employed the Dual‐Luciferase Reporter Assay System and Electrophoretic Mobility Shift Assay (EMSA) and demonstrated that MYOD, a major myogenic transcription factor, binds directly to the promoter region of Fgf21, leading to the activation of Fgf21 expression in mouse C2C12 myoblasts. Our study revealed a novel mechanism of myogenesis and muscle fiber transformation and indicated that FGF21 serves as a vital regulator of muscle development and important contributor to the pathogenesis of myopathy. J. Cell. Physiol. 232: 1893–1906, 2017. © 2016 Wiley Periodicals, Inc. (1) The mouse FGF21 gene expression is positively regulated by the transcription factor MyoD. (2) FGF21 promotes myogenic differentiation and facilitates the switch of the muscle fiber type from anaerobic myofibers to aerobic myofibers via stimulation the FGF21‐SIRT1‐AMPK‐PGC1α axis. (3) The increased FGF21 protein induced by fasting is likely to change the expression of the PGC‐1α protein from a physiological to an abnormally excessive level via the activation of AMPK signaling, which results in muscle atrophy.

  January 05, 2017   doi: 10.1002/jcp.25735   open full text
• Roles of Notch Signaling in Adipocyte Progenitor Cells and Mature Adipocytes.
  Tizhong Shan, Jiaqi Liu, Weiche Wu, Ziye Xu, Yizhen Wang.
  Journal of Cellular Physiology. January 05, 2017
  Adipose tissues, composed with mature adipocytes and preadipocytic stromal/stem cells, play crucial roles in whole body energy metabolism and regenerative medicine. Mature adipocytes are derived and differentiated from mesenchymal stem cells (MSCs) or preadipocytes. This differentiation process, also called adipogenesis, is regulated by several signaling pathways and transcription factors. Notch1 signaling is a highly conserved pathway that is indispensable for stem cell hemostasis and tissue development. In adipocyte progenitor cells, Notch1 signaling regulates the adipogenesis process including proliferation and differentiation of the adipocyte progenitor cells in vitro. Notably, the roles of Notch1 signaling in beige adipocytes formation, adipose development, and function, and the whole body energy metabolism have been recently reported. Here, we mainly review and discuss the roles of Notch1 signaling in adipogenesis in vitro as well as in beige adipocytes formation, adipocytes dedifferentiation, and function in vivo. J. Cell. Physiol. 232: 1258–1261, 2017. © 2016 Wiley Periodicals, Inc. Notch1 signaling not only regulates the adipogenesis process including proliferation and differentiation of the adipocyte progenitor cells in vitro, but also affects beige adipocytes formation, adipose development, and function, and the whole body energy metabolism in vivo.

  January 05, 2017   doi: 10.1002/jcp.25697   open full text
• Modulation of Chloride Currents in Human Lung Epithelial Cells Exposed to Exogenous Oxidative Stress.
  Rita Canella, Marta Martini, Roberta Borriello, Carlotta Cavicchio, Ximena M. Muresan, Mascia Benedusi, Franco Cervellati, Giuseppe Valacchi.
  Journal of Cellular Physiology. January 05, 2017
  Air pollution continues to be a major public health concern affecting 9 out of 10 individuals living in urban areas worldwide. Respiratory tract is the organ most exposed to gas pollution, and ozone has been shown to be one of the most noxious pollutants to which living organisms are exposed. In the present work, we have investigated the effects of 0.1 ppm of ozone on chloride currents in human lung epithelial cells (A549 line) and whether this effect could be modulated by vitamin E pre‐treatment. Whole‐cell patch clamp technique was applied to not excitable cells in order to obtain information about chloride currents behavior, important for epithelial lung cells homeostasis. Significant alteration of the I–V curve after ozone treatment was observed, with the appearance of a large outward rectifier component decreasing over time and returning to the basal state levels after 24 h. Statistical analysis indicated a modification of the amount of ions passing the membrane in the unit of time as a possible cause of this difference. RT‐qPCR analysis showed an increase in ClC‐2 and ORCC mRNA after ozone exposure. In addition, pre‐treatment with vitamin E was able to suppress the outward rectifier component induced by ozone, bringing back the current values to the control level and preventing ozone induced chloride channels up regulation. Our data suggest that ozone exposure is able to modify chloride current density and the use of vitamin E can prevent the above‐mentioned damage. J. Cell. Physiol. 232: 1817–1825, 2017. © 2016 Wiley Periodicals, Inc. Pollutant ozone is able to induce oxidative stress to lung epithelial cells and this affect chloride currents and the expression of ClC2 and ORCC channel genes levels. Vitamin E pre‐treatment is able to rescue this noxious effect.

  January 05, 2017   doi: 10.1002/jcp.25705   open full text
• A Human 3D In Vitro Model to Assess the Relationship Between Osteoporosis and Dissemination to Bone of Breast Cancer Tumor Cells.
  Francesca Salamanna, Veronica Borsari, Silvia Brogini, Paola Torricelli, Simona Cepollaro, Matteo Cadossi, Milena Fini.
  Journal of Cellular Physiology. January 05, 2017
  Despite consistent improvements in diagnostic and therapeutic strategies for breast cancer, up to 40% of patients will develop bone metastases. To reduce the morbidity and complications related with bone metastases, it is imperative to reduce their etiological factors. Osteoporosis, being characterized by a sudden estrogen deficiency, may provide a favorable condition for bone metastasis. This work, using a humanized 3D in vitro model, aims at evaluating the relationship between osteoporosis and breast cancer‐derived bone metastases. Bone tissue discarded from total hip replacement surgery of healthy and osteoporotic patients was cultured in a rolling apparatus system in hypoxic environment. Protein levels (i.e., vascular endothelial growth factor (VEGF), VEGF receptor 1, VEGF receptor 2, interleukin (IL)‐6, IL‐1β, IL‐8 IL‐10, tumor necrosis factor α (TNF‐α), osteoprotegerin (OPG), receptor activator for nuclear factor KB ligand (RANKL)) and histological and immunohistochemical (i.e., cytokeratin 8 and 18) analyses showed a noticeable specificity of breast cancer cells for the colonization of osteoporotic bone. These data are the first to demonstrate that using humanized 3D in vitro systems, which individually model the pre‐ and postmenopausal bone microenvironment, it is possible to recognize major differences in tumor growth and colonization between healthy and osteoporotic status. Thus, this system might help to develop a shared system between basic and clinical sciences where a personalized diagnosis is associated to a therapeutic strategy designed for a single patient: a model able to achieve a translational research approach in the clinical setting, which may lead to the application and dissemination of personalized medicine. J. Cell. Physiol. 232: 1826–1834, 2017. © 2016 Wiley Periodicals, Inc. Humanized 3D in vitro model; relationship between osteoporosis and breast cancer‐derived bone metastases; differences in tumor growth and colonization between healthy and osteoporotic status.

  January 05, 2017   doi: 10.1002/jcp.25708   open full text
• Identifying Nuclear Matrix‐Attached DNA Across the Genome.
  Jason R. Dobson, Deli Hong, A. Rasim Barutcu, Hai Wu, Anthony N. Imbalzano, Jane B. Lian, Janet L. Stein, Andre J. van Wijnen, Jeffrey A. Nickerson, Gary S. Stein.
  Journal of Cellular Physiology. January 05, 2017
  Experimental approaches to define the relationship between gene expression and nuclear matrix attachment regions (MARs) have given contrasting and method‐specific results. We have developed a next generation sequencing strategy to identify MARs across the human genome (MAR‐Seq). The method is based on crosslinking chromatin to its nuclear matrix attachment sites to minimize changes during biochemical processing. We used this method to compare nuclear matrix organization in MCF‐10A mammary epithelial‐like cells and MDA‐MB‐231 breast cancer cells and evaluated the results in the context of global gene expression (array analysis) and positional enrichment of gene‐regulatory histone modifications (ChIP‐Seq). In the normal‐like cells, nuclear matrix‐attached DNA was enriched in expressed genes, while in the breast cancer cells, it was enriched in non‐expressed genes. In both cell lines, the chromatin modifications that mark transcriptional activation or repression were appropriately associated with gene expression. Using this new MAR‐Seq approach, we provide the first genome‐wide characterization of nuclear matrix attachment in mammalian cells and reveal that the nuclear matrix‐associated genome is highly cell‐context dependent. J. Cell. Physiol. 232: 1295–1305, 2017. © 2016 Wiley Periodicals, Inc. MAR‐Seq is a next‐generation sequencing strategy to identify matrix attachment regions (MARs) that is based on crosslinking chromatin to its nuclear attachment sites. Comparison of MCF‐10A mammary epithelial‐like cells and MDA‐MB‐231 breast cancer cells showed that nuclear matrix‐attached DNA in normal‐like MCF‐10A cells is enriched in expressed genes, while in the breast cancer cells, it is enriched in non‐expressed genes. This new MAR‐Seq approach reveals that the nuclear matrix‐associated genome is highly cell‐context dependent.

  January 05, 2017   doi: 10.1002/jcp.25596   open full text
• Primary Cilium‐Regulated EG‐VEGF Signaling Facilitates Trophoblast Invasion.
  Chia‐Yih Wang, Hui‐Ling Tsai, Jhih‐Siang Syu, Ting‐Yu Chen, Mei‐Tsz Su.
  Journal of Cellular Physiology. December 29, 2016
  Trophoblast invasion is an important event in embryo implantation and placental development. During these processes, endocrine gland‐derived vascular endothelial growth factor (EG‐VEGF) is the key regulator mediating the crosstalk at the feto‐maternal interface. The primary cilium is a cellular antenna receiving environmental signals and is crucial for proper development. However, little is known regarding the role of the primary cilium in early human pregnancy. Here, we demonstrate that EG‐VEGF regulates trophoblast cell invasion via primary cilia. We found that EG‐VEGF activated ERK1/2 signaling and subsequent upregulation of MMP2 and MMP9, thereby facilitating cell invasion in human trophoblast HTR‐8/SVneo cells. Inhibition of ERK1/2 alleviated the expression of MMPs and trophoblast cell invasion after EG‐VEGF treatment. In addition, primary cilia were observed in all the trophoblast cell lines tested and, more importantly, in human first‐trimester placental tissue. The receptor of EG‐VEGF, PROKR1, was detected in primary cilia. Depletion of IFT88, the intraflagellar transporter required for ciliogenesis, inhibited primary cilium growth, thereby ameliorating ERK1/2 activation, MMP upregulation, and trophoblast cell invasion promoted by EG‐VEGF. These findings demonstrate a novel function of primary cilia in controlling EG‐VEGF‐regulated trophoblast invasion and reveal the underlying molecular mechanism. J. Cell. Physiol. 232: 1467–1477, 2017. © 2016 Wiley Periodicals, Inc. Primary cilium is a cellular antenna that conveys extracellular signals into a cellular response. EG‐VEGF binds to PROKR1 locating on the cilium, and activates ERK1/2 signaling followed by upregulating MMP2 and MMP9 for facilitating cell invasion. We demonstrated that EG‐VEGF regulated trophoblast cell invasion via primary cilium in the present study.

  December 29, 2016   doi: 10.1002/jcp.25649   open full text
• Identification of a Novel Human E‐Cadherin Splice Variant and Assessment of Its Effects Upon EMT‐Related Events.
  María Laura Matos, Lara Lapyckyj, Marina Rosso, María José Besso, María Victoria Mencucci, Clara Isabel Marín Briggiler, Silvina Giustina, Laura Inés Furlong, Mónica Hebe Vazquez‐Levin.
  Journal of Cellular Physiology. December 29, 2016
  Epithelial Cadherin (E‐cadherin) is involved in calcium‐dependent cell–cell adhesion and signal transduction. The E‐cadherin decrease/loss is a hallmark of Epithelial to Mesenchymal Transition (EMT), a key event in tumor progression. The underlying molecular mechanisms that trigger E‐cadherin loss and consequent EMT have not been completely elucidated. This study reports the identification of a novel human E‐cadherin variant mRNA produced by alternative splicing. A bioinformatics evaluation of the novel mRNA sequence and biochemical verifications suggest its regulation by Nonsense‐Mediated mRNA Decay (NMD). The novel E‐cadherin variant was detected in 29/42 (69%) human tumor cell lines, expressed at variable levels (E‐cadherin variant expression relative to the wild type mRNA = 0.05–11.6%). Stable transfection of the novel E‐cadherin variant in MCF‐7 cells (MCF7Ecadvar) resulted in downregulation of wild type E‐cadherin expression (transcript/protein) and EMT‐related changes, among them acquisition of a fibroblastic‐like cell phenotype, increased expression of Twist, Snail, Zeb1, and Slug transcriptional repressors and decreased expression of ESRP1 and ESRP2 RNA binding proteins. Moreover, loss of cytokeratins and gain of vimentin, N‐cadherin and Dysadherin/FXYD5 proteins was observed. Dramatic changes in cell behavior were found in MCF7Ecadvar, as judged by the decreased cell–cell adhesion (Hanging‐drop assay), increased cell motility (Wound Healing) and increased cell migration (Transwell) and invasion (Transwell w/Matrigel). Some changes were found in MCF‐7 cells incubated with culture medium supplemented with conditioned medium from HEK‐293 cells transfected with the E‐cadherin variant mRNA. Further characterization of the novel E‐cadherin variant will help understanding the molecular basis of tumor progression and improve cancer diagnosis. J. Cell. Physiol. 232: 1368–1386, 2017. © 2016 Wiley Periodicals, Inc. This report describes the identification and partial characterization of a novel human E‐cadherin variant mRNA produced by alternative splicing. Stable transfection of the E‐cadherin variant mRNA in MCF‐7 cells (MCF7Ecadvar cells) resulted in downregulation of wild type E‐cadherin and EMT‐related changes, that is, acquisition of a fibroblast‐like cellular phenotype, increased expression of E‐cadherin transcriptional repressors, vimentin and N‐cadherin, and loss of cytokeratins. MCF7Ecadvar cells depicted dramatic changes in cellular behavior, showing reduced cell–cell adhesion and increased cell migration and invasiveness.

  December 29, 2016   doi: 10.1002/jcp.25622   open full text
• Exploring the Role of PGC‐1α in Defining Nuclear Organisation in Skeletal Muscle Fibres.
  Jacob Alexander Ross, Adam Pearson, Yotam Levy, Bettina Cardel, Christoph Handschin, Julien Ochala.
  Journal of Cellular Physiology. December 29, 2016
  Muscle fibres are multinucleated cells, with each nucleus controlling the protein synthesis in a finite volume of cytoplasm termed the myonuclear domain (MND). What determines MND size remains unclear. In the present study, we aimed to test the hypothesis that the level of expression of the transcriptional coactivator PGC‐1α and subsequent activation of the mitochondrial biogenesis are major contributors. Hence, we used two transgenic mouse models with varying expression of PGC‐1α in skeletal muscles. We isolated myofibres from the fast twitch extensor digitorum longus (EDL) and slow twitch diaphragm muscles. We then membrane‐permeabilised them and analysed the 3D spatial arrangements of myonuclei. In EDL muscles, when PGC‐1α is over‐expressed, MND volume decreases; whereas, when PGC‐1α is lacking, no change occurs. In the diaphragm, no clear difference was noted. This indicates that PGC‐1α and the related mitochondrial biogenesis programme are determinants of MND size. PGC‐1α may facilitate the addition of new myonuclei in order to reach MND volumes that can support an increased mitochondrial density. J. Cell. Physiol. 232: 1270–1274, 2017. © 2016 Wiley Periodicals, Inc. Muscle fibres are multinucleated cells, with individual nuclei controlling the protein synthesis in a finite volume of cytoplasm termed myonuclear domain. The present study indicates that one important regulator of myonuclear domain size is PGC‐1α and related mitochondrial biogenesis programme.

  December 29, 2016   doi: 10.1002/jcp.25678   open full text
• SV40 Infection of Mesenchymal Stromal Cells From Wharton's Jelly Drives the Production of Inflammatory and Tumoral Mediators.
  Carolina Cason, Giuseppina Campisciano, Nunzia Zanotta, Erica Valencic, Serena Delbue, Ramona Bella, Manola Comar.
  Journal of Cellular Physiology. December 29, 2016
  The Mesenchymal Stromal Cells from umbilical cord Wharton's jelly (WJSCs) are a source of cells with high potentiality for the treatment of human immunological disorders. Footprints of the oncogenic viruses Simian Virus 40 (SV40) and JC Virus (JCPyV) have been recently detected in human WJSCs specimens. The aim of this study is to evaluate if WJSCs can be efficiently infected by these Polyomaviruses and if they can potentially exert tumoral activity. Cell culture experiments indicated that WJSCs could sustain both SV40 and JCPyV infections. A transient and lytic replication was observed for JCPyV, while SV40 persistently infected WJSCs over a long period of time, releasing a viral progeny at low titer without evident cytopathic effect (CPE). Considering the association between SV40 and human tumors and the reported ability of the oncogenic viruses to drive the host innate immune response to cell transformation, the expression profile of a large panel of immune mediators was evaluated in supernatants by the Bioplex platform. RANTES, IL‐3, MIG, and IL‐12p40, involved in chronic inflammation, cells differentiation, and transformation, were constantly measured at high concentration comparing to control. These findings represent a new aspect of SV40 biological activity in the humans, highlighting its interaction with specific host cellular pathways. In view of these results, it seems to be increasingly urgent to consider Polyomaviruses in the management of WJSCs for their safely use as promising therapeutic source. J. Cell. Physiol. 9999: 1–7, 2016. © 2016 Wiley Periodicals, Inc. This study aims to evaluate if WJSCs can be efficiently infected by the Polyomaviruses SV40 and JCPyV and if they can exert their potential tumoral activity. Results reported a transient and lytic replication for JCPyV, while SV40 persistently infected WJSCs over a long period of time, releasing viral progeny. Specific factors, including RANTES, IL‐3, MIG, and IL‐12p40, involved in chronic inflammation, cells differentiation, and transformation, were constantly produced at high concentration. These findings represent a new aspect of SV40 biological activity in human host. In view of these results, it seems to be increasingly urgent to consider Polyomaviruses in the management of WJSCs for their safely use as promising therapeutic source.

  December 29, 2016   doi: 10.1002/jcp.25723   open full text
• Lgr4 Expression in Osteoblastic Cells Is Suppressed by Hydrogen Peroxide Treatment.
  Chantida Pawaputanon Na Mahasarakham, Yayoi Izu, Katsuhiko Nishimori, Yuichi Izumi, Masaki Noda, Yoichi Ezura.
  Journal of Cellular Physiology. December 29, 2016
  LGR4 is expressed in bone and has been shown to be involved in bone metabolism. Oxidative stress is one of the key issues in pathophysiology of osteoporosis. However, the link between Lgr4 and oxidative stress has not been known. Therefore, effects of hydrogen peroxide on Lgr4 expression in osteoblasts were examined. Hydrogen peroxide treatment suppressed the levels of Lgr4 mRNA expression in an osteoblastic cell line, MC3T3‐E1. The suppressive effects were not obvious at 0.1 mM, while 1 mM hydrogen peroxide suppressed Lgr4 expression by more than 50%. Hydrogen peroxide treatment suppressed Lgr4 expression within 12 h and this suppression lasted at least up to 48 h. Hydrogen peroxide suppression of Lgr4 expression was still observed in the presence of a transcription inhibitor but was no longer observed in the presence of a protein synthesis inhibitor. Although Lgr4 expression in osteoblasts is enhanced by BMP2 treatment as reported before, hydrogen peroxide treatment suppressed Lgr4 even in the presence of BMP2. Finally, hydrogen peroxide suppressed Lgr4 expression in primary cultures of osteoblasts similarly to MC3T3‐E1 cells. These date indicate that hydrogen peroxide suppresses Lgr4 expression in osteoblastic cells. J. Cell. Physiol. 232: 1761–1766, 2017. © 2016 Wiley Periodicals, Inc.

  December 29, 2016   doi: 10.1002/jcp.25684   open full text
• Diverse Functions and Signal Transduction of the Exocyst Complex in Tumor Cells.
  Toshiaki Tanaka, Kaoru Goto, Mitsuyoshi Iino.
  Journal of Cellular Physiology. December 22, 2016
  The exocyst complex is a large conserved hetero‐oligomeric complex that consists of Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84 subunits. It has been implicated in the targeting of vesicles for regulated exocytosis in various cell types, and is also important for targeted exocytosis of post‐Golgi transport vesicles to the plasma membrane. The exocyst complex is essential for membrane growth, secretion, and function during exocytosis and endocytosis. Moreover, the individual components of the complex are thought to act on specific biological processes, such as cytokinesis, ciliogenesis, apoptosis, autophagy, and epithelial‐mesenchymal transition (EMT). As a result, recent studies suggest that the exocyst complex may be involved in several diseases such as kidney disease, neuropathogenesis, diabetes, and cancer. In this review, we focus on the diverse functions and cellular signaling pathways of the exocyst complex in various tumors. J. Cell. Physiol. 232: 939–957, 2017. © 2016 Wiley Periodicals, Inc. Exo70 interacts strongly with the Sec10–Sec15 subcomplex and Exo84 interacts with the Sec10–Sec15–Exo70 subcomplex. Sec3, Sec5, Sec6, and Sec8 form a tight quaternary subcomplex and Sec8 interacts with Sec10. Each component of the exocyst complex contributes to cancer progression and survival in diverse ways such as break of cell–cell contact, EMT, cell cycle progression, DNA repair, migration, autophagy, and anti‐apoptosis.

  December 22, 2016   doi: 10.1002/jcp.25619   open full text
• Differential Expression of miR‐4520a Associated With Pyrin Mutations in Familial Mediterranean Fever (FMF).
  Helen Latsoudis, Mir‐Farzin Mashreghi, Joachim R. Grün, Hyun‐Dong Chang, Bruno Stuhlmüller, Argyro Repa, Irini Gergiannaki, Eleni Kabouraki, George S. Vlachos, Thomas Häupl, Andreas Radbruch, Prodromos Sidiropoulos, Kimon Doukoumetzidis, Dimitris Kardassis, Timothy B. Niewold, Dimitrios T. Boumpas, George N. Goulielmos.
  Journal of Cellular Physiology. December 20, 2016
  Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute, and self‐limiting attacks of fever. Mutations in MEFV gene encoding pyrin account for FMF, but the high number of heterozygote patients with typical symptoms of the disease has driven a number of alternative aetiopathogenic hypotheses. The MEFV gene was knocked down in human myelomonocytic cells that express endogenous pyrin to identify deregulated microRNAs (miRNAs). Microarray analyses revealed 29 significantly differentially expressed miRNAs implicated in pathways associated with cellular integrity and survival. Implementation of in silico gene network prediction algorithms and bioinformatics analyses showed that miR‐4520a is predicted to target genes implicated in autophagy through regulation of RHEB/mTOR signaling. Differential expression levels of RHEB were confirmed by luciferase reporter gene assays providing further evidence that is directly targeted by miR‐4520a. Although the relative expression levels of miR‐4520a were variable among FMF patients, the statistical expression of miR‐4520a was different between FMF mutation carriers and controls (P = 0.0061), indicating an association between miR‐4520a expression and MEFV mutations. Comparison between FMF patients bearing the M694V mutation, associated with severe disease, and healthy controls showed a significant increase in miR‐4520a expression levels (P = 0.00545). These data suggest that RHEB, the main activator of mTOR signaling, is a valid target of miR‐4520a with the relative expression levels of the latter being significantly deregulated in FMF patients and highly dependent on the presence of pyrin mutations, especially of the M694V type. These results suggest a role of deregulated autophagy in the pathogenesis of FMF. J. Cell. Physiol. 232: 1326–1336, 2017. © 2016 Wiley Periodicals, Inc. Familial Mediterranean fever (FMF) is an autosomal recessive disease characterized by recurrent, acute and self‐limiting attacks of fever. In the present study we presented for the first time results supporting a role of deregulated autophagy in the pathogenesis of FMF.

  December 20, 2016   doi: 10.1002/jcp.25602   open full text
• Overexpression of Gremlin1 in Mesenchymal Stem Cells Improves Hindlimb Ischemia in Mice by Enhancing Cell Survival.
  Qiuling Xiang, Dongxi Hong, Yan Liao, Yong Cao, Muyun Liu, Jun Pang, Junjie Zhou, Guang Wang, Renhao Yang, Maosheng Wang, Andy Peng Xiang.
  Journal of Cellular Physiology. December 20, 2016
  Mesenchymal stem cells (MSCs) are a promising cell resource for the treatment of ischemic diseases, partially through paracrine effects. One of the major obstacles of MSC treatment is the poor survival rate and low efficiency of transplanted stem cells due to ischemic or inflammatory environments. Gremlin1 (GREM1), a regulator of growth, differentiation and development, has been identified as a novel proangiogenic factor. However, the role and mechanism of GREM1 in MSCs remains unclear. Therefore, we assessed the putative beneficial effects of GREM1 on MSC‐based therapy for hindlimb ischemia. The lentiviral vector, EF1a‐GREM1, was constructed using the Multisite Gateway System and used to transduce MSCs. In vitro studies demonstrated increased survival of GREM1‐MSCs exposed to H2O2, which is consistent with the activation of caspase‐3. Conditional medium from GREM1‐MSCs (GREM1‐MSC‐CM) increased the anti‐apoptotic effects of human umbilical vein endothelial cells (HUVECs), and this effect was attenuated by treatment with the PI3K/Akt pathway inhibitor LY294002. MSCs modified with GREM1 could significantly increase blood perfusion of the ischemic hindlimb in vivo in a mouse model, which was correlated to improved MSC survival. This study demonstrates that overexpression of GREM1 in MSCs have greater therapeutic effects against ischemia compared with wild‐type MSCs by enhancing the survival of MSCs and ECs, which may provide new tools for studies investigating the treatment of ischemic diseases. J. Cell. Physiol. 232: 996–1007, 2017. © 2016 Wiley Periodicals, Inc. This study demonstrates that overexpression of GREM1 in MSCs have greater therapeutic effects against ischemia compared with wild‐type MSCs by enhancing the survival of MSCs and ECs, which may provide new tools for studies investigating the treatment of ischemic diseases.

  December 20, 2016   doi: 10.1002/jcp.25578   open full text
• Supernatants of Adipocytes From Obese Versus Normal Weight Women and Breast Cancer Cells: In Vitro Impact on Angiogenesis.
  Lauriane Bougaret, Laetitia Delort, Hermine Billard, Charlotte Lequeux, Nicolas Goncalves‐Mendes, Ali Mojallal, Odile Damour, Marie‐Paule Vasson, Florence Caldefie‐Chezet.
  Journal of Cellular Physiology. December 20, 2016
  Breast cancer is correlated with a higher risk of metastasis in obese postmenopausal women. Adipokines, whose plasma concentrations are modulated in obese subjects and adipocytes surround mammary cells, suggesting that adipocyte secretome affect mammary tumorogenesis. We hypothesize that mature adipocyte secretions from obese women conditioned or not by breast neoplasic cells, increase changes on the angiogenesis stages. Supernatants of human mature adipocytes, differentiated from stem cells of either adipose tissue of normal weight (MA20) or obese (MA30) women or obtained from co‐cultures between MA20 and MA30 and breast cancer cell line MCF‐7, were collected. The impact of these supernatants was investigated on proliferation, migration, and tube formation by endothelial cells (HUVEC). MA20 and MA30 showed a preservation of their “metabolic memory” (increase of Leptin, ObR, VEGF, CYP19A1, and a decrease of Adiponectin expression in MA30 compared to MA20). Supernatants from obese‐adipocytes increased HUVEC proliferation, migration, and sprouting like with supernatants obtained from co‐cultures of MA/MCF‐7 regardless the women's BMI. Additional analyses such as the use of neutralizing antibodies, analysis of supernatants (Milliplex®) and variations in gene expression (qRT‐PCR), strongly suggest an implication of IL‐6, or a synergistic action among adipokines, probably associated with that of VEGF or IL‐6. As a conclusion, supernatants from co‐cultures of MA30 and MCF‐7 cells increase proliferation, migration, and sprouting of HUVEC cells. These results provide insights into the interaction between adipocytes and epithelial cancer cells, particularly in case of obesity. The identification of synergistic action of adipokines would therefore be a great interest in developing preventive strategies. J. Cell. Physiol. 232: 1808–1816, 2017. © 2016 Wiley Periodicals, Inc. We characterized the impact of adipokines from obese or normal weight women on different angiogenesis stages (proliferation, migration, and tube formation of endothelial cells) in order to determine the influence of obesity on this process and the impact of adipocytes on breast tumor cells.

  December 20, 2016   doi: 10.1002/jcp.25701   open full text
• Acute Ethanol Increases IGF‐I‐Induced Phosphorylation of ERKs by Enhancing Recruitment of p52‐Shc to the Grb2/Shc Complex.
  Matthew Dean, Adam Lassak, Anna Wilk, Adriana Zapata, Luis Marrero, Patricia Molina, Krzysztof Reiss.
  Journal of Cellular Physiology. December 19, 2016
  Ethanol plays a detrimental role in the development of the brain. Multiple studies have shown that ethanol inhibits insulin‐like growth factor I receptor (IGF‐IR) function. Because the IGF‐IR contributes to brain development by supporting neural growth, survival, and differentiation, we sought to determine the molecular mechanism(s) involved in ethanol's effects on this membrane‐associated tyrosine kinase. Using multiple neuronal cell types, we performed Western blot, immunoprecipitation, and GST‐pulldowns following acute (1–24 h) or chronic (3 weeks) treatment with ethanol. Surprisingly, exposure of multiple neuronal cell types to acute (up to 24 h) ethanol (50 mM) enhanced IGF‐I‐induced phosphorylation of extracellular regulated kinases (ERKs), without affecting IGF‐IR tyrosine phosphorylation itself, or Akt phosphorylation. This acute increase in ERKs phosphorylation was followed by the expected inhibition of the IGF‐IR signaling following 3‐week ethanol exposure. We then expressed a GFP‐tagged IGF‐IR construct in PC12 cells and used them to perform fluorescence recovery after photobleaching (FRAP) analysis. Using these fluorescently labeled cells, we determined that 50 mM ethanol decreased the half‐time of the IGF‐IR‐associated FRAP, which implied that cell membrane‐associated signaling events could be affected. Indeed, co‐immunoprecipitation and GST‐pulldown studies demonstrated that the acute ethanol exposure increased the recruitment of p52‐Shc to the Grb2‐Shc complex, which is known to engage the Ras‐Raf‐ERKs pathway following IGF‐1 stimulation. These experiments indicate that even a short and low‐dose exposure to ethanol may dysregulate function of the receptor, which plays a critical role in brain development. J. Cell. Physiol. 232: 1275–1286, 2017. © 2016 Wiley Periodicals, Inc. In the current study, we show a novel effect of ethanol on the IGF‐I receptor (IGF‐IR) signaling, and provided a possible mechanism through which this occurs. We determine that ethanol enhances mobility of the IGF‐IR molecules within the cell membrane, and that this stimulatory effect enhances formation of the Grb2‐Shc signaling complex leading to the transient increase of the signal toward extracellular regulated kinases (ERKs). Because of the critical role of the IGF‐IR signaling in neuronal growth and survival, and the detrimental effects of alcohol on CNS development, we believe that our new data will help in better understanding of the fetal alcohol syndrome.

  December 19, 2016   doi: 10.1002/jcp.25586   open full text
• Association Between Aerobic Exercise and Rosiglitazone Avoided the NAFLD and Liver Inflammation Exacerbated in PPAR‐α Knockout Mice.
  Helena A.P. Batatinha, Edson A. Lima, Alexandre A.S. Teixeira, Camila O. Souza, Luana A. Biondo, Loreana S. Silveira, Fabio S. Lira, José C. Rosa Neto.
  Journal of Cellular Physiology. November 30, 2016
  Nonalcoholic fatty liver disease (NAFLD) is one of the main liver diseases today, and may progress to steatohepatitis, cirrhosis, and hepatocellular carcinoma. Some studies have shown the beneficial effects of aerobic exercise on reversing NAFLD. To verify whether chronic aerobic exercise improves the insulin resistance, liver inflammation, and steatohepatitis caused by a high fat diet (HF) and whether PPARα is involved in these actions. C57BL6 wild type (WT) and PPAR‐α knockout (KO) mice were fed with a standard diet (SD) or HF during 12 weeks; the HF mice were trained on a treadmill during the last 8 weeks. Serum glucose and insulin tolerances, serum levels of aspartate aminotransferase, hepatic content of triacylglycerol, cytokines, gene expression, and protein expression were evaluated in all animals. Chronic exposure to HF diet increased triacylglycerol accumulation in the liver, leading to NAFLD, increased aminotransferase in the serum, increased peripheral insulin resistance, and higher adiposity index. Exercise reduced all these parameters in both animal genotypes. The liver lipid accumulation was not associated with inflammation; trained KO mice, however, presented a huge inflammatory response that was probably caused by a decrease in PPAR‐γ expression. We conclude that exercise improved the damage caused by a HF independently of PPARα, apparently by a peripheral fatty acid oxidation in the skeletal muscle. We also found that the absence of PPARα together with exercise leads to a decrease in PPAR‐γ and a huge inflammatory response. J. Cell. Physiol. 232: 1008–1019, 2017. © 2016 Wiley Periodicals, Inc. Exercise training improved lipids toxicity in the liver; PPAR‐alpha KO developed liver inflammation after training protocol Roziglitazone reduced liver inflammation independent of PPAR‐alpha.

  November 30, 2016   doi: 10.1002/jcp.25440   open full text
• PPARα Antagonist AA452 Triggers Metabolic Reprogramming and Increases Sensitivity to Radiation Therapy in Human Glioblastoma Primary Cells.
  Elisabetta Benedetti, Michele d'Angelo, Alessandra Ammazzalorso, Giovanni Luca Gravina, Chiara Laezza, Andrea Antonosante, Gloria Panella, Benedetta Cinque, Loredana Cristiano, Anne Chloè Dhez, Carlo Astarita, Renato Galzio, Maria Grazia Cifone, Rodolfo Ippoliti, Rosa Amoroso, Ernesto Di Cesare, Antonio Giordano, Annamaria Cimini.
  Journal of Cellular Physiology. November 30, 2016
  Glioblastoma (GB) is the most common cancer in the brain and with an increasing incidence. Despite major advances in the field, there is no curative therapy for GB to date. Many solid tumors, including GB, experienced metabolic reprogramming in order to sustain uncontrolled proliferation, hypoxic conditions, and angiogenesis. PPARs, member of the steroid hormone receptor superfamily, are particularly involved in the control of energetic metabolism, particularly lipid metabolism, which has been reported deregulated in gliomas. PPARα was previously indicated by us as a potential therapeutic target for this neoplasm, due to the malignancy grade dependency of its expression, being particularly abundant in GB. In this work, we used a new PPARα antagonist on patient‐derived GB primary cells, with particular focus on the effects on lipid metabolism and response to radiotherapy. The results obtained demonstrated that blocking PPARα results in cell death induction, increase of radiosensitivity, and decrease of migration. Therefore, AA452 is proposed as a new adjuvant for the gold standard therapies for GB, opening the possibility for preclinical and clinical trials for this class of compounds. J. Cell. Physiol. 232: 1458–1466, 2017. © 2016 Wiley Periodicals, Inc. A new compound, an inhibitor of PPARalpha trancriptional activity, is reported, able to increase radiation sensitivity of glioblastoma cells.

  November 30, 2016   doi: 10.1002/jcp.25648   open full text
• Inhibitory Effects of Quercetin on Progression of Human Choriocarcinoma Cells Are Mediated Through PI3K/AKT and MAPK Signal Transduction Cascades.
  Whasun Lim, Changwon Yang, Sunwoo Park, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. November 30, 2016
  As a major dietary flavonol, quercetin mitigates proliferation and progression of cancer due to its anti‐angiogenic, anti‐inflammatory, anti‐oxidant, and apoptotic biological effects on cells. Although its apoptotic effects have been reported for various cancers, little is known of the functional role of quercetin in gestational choriocarcinoma. Results of the present study indicated that quercetin reduced proliferation and induced cell death in two choriocarcinoma cell lines, JAR and JEG3 cells, with an increase in the sub‐G1 phase of the cell cycle. In addition, quercetin induced mitochondrial dysfunction significantly reduced mitochondrial membrane potential (MMP) and increased production of reactive oxygen species (ROS) in both JAR and JEG3 cells. Further, quercetin inhibited phosphorylation of AKT, P70S6K and S6 proteins whereas, it increased phosphorylation of ERK1/2, P38, JNK and P90RSK proteins in JAR and JEG3 cells. The decrease in viability of choriocarcinoma cells treated with quercetin was confirmed by using combinations of quercetin and pharmacological inhibitors of the PI3K and MAPK signaling pathways. Classical chemotherapeutic agents, cisplatin (a platinum‐based drug) and paclitaxel (a taxene‐based drug), inhibited proliferation of JAR and JEG3 cells, and when combined with quercetin, the antiproliferative effects of cisplatin and paclitaxel were enhanced for both choriocarcinoma cell lines. Collectively, these results suggest that quercetin prevents development of choriocarcinoma and may be a valuable therapeutic agent for treatment of choriocarcinoma through its regulation of PI3K and MAPK signal transduction pathways. J. Cell. Physiol. 232: 1428–1440, 2017. © 2016 Wiley Periodicals, Inc. Quercetin prevents development of choriocarcinoma and may be a valuable therapeutic agent for treatment of choriocarcinoma through its regulation of PI3K/AKT and MAPK signaling pathways and intrinsic apoptotic pathways.

  November 30, 2016   doi: 10.1002/jcp.25637   open full text
• TRPV4 Regulates Tight Junctions and Affects Differentiation in a Cell Culture Model of the Corneal Epithelium.
  Jacqueline Martínez‐Rendón, Erika Sánchez‐Guzmán, Angélica Rueda, James González, Rosario Gulias‐Cañizo, Guillermo Aquino‐Jarquín, Federico Castro‐Muñozledo, Refugio García‐Villegas.
  Journal of Cellular Physiology. November 30, 2016
  TRPV4 (transient receptor potential vanilloid 4) is a cation channel activated by hypotonicity, moderate heat, or shear stress. We describe the expression of TRPV4 during the differentiation of a corneal epithelial cell model, RCE1(5T5) cells. TRPV4 is a late differentiation feature that is concentrated in the apical membrane of the outmost cell layer of the stratified epithelia. Ca2+ imaging experiments showed that TRPV4 activation with GSK1016790A produced an influx of calcium that was blunted by the specific TRPV4 blocker RN‐1734. We analyzed the involvement of TRPV4 in RCE1(5T5) epithelial differentiation by measuring the development of transepithelial electrical resistance (TER) as an indicator of the tight junction (TJ) assembly. We showed that TRPV4 activity was necessary to establish the TJ. In differentiated epithelia, activation of TRPV4 increases the TER and the accumulation of claudin‐4 in cell–cell contacts. Epidermal Growth Factor (EGF) up‐regulates the TER of corneal epithelial cultures, and we show here that TRPV4 activation mimicked this EGF effect. Conversely, TRPV4 inhibition or knock down by specific shRNA prevented the increase in TER. Moreover, TRPP2, an EGF‐activated channel that forms heteromeric complexes with TRPV4, is also concentrated in the outmost cell layer of differentiated RCE1(5T5) sheets. This suggests that the EGF regulation of the TJ may involve a heterotetrameric TRPV4‐TRPP2 channel. These results demonstrated TRPV4 activity was necessary for the correct establishment of TJ in corneal epithelia and as well as the regulation of both the barrier function of TJ and its ability to respond to EGF. J. Cell. Physiol. 232: 1794–1807, 2017. © 2016 Wiley Periodicals, Inc. TRPV4 channel activity is necessary for the correct establishment of tight Junctions (TJ) in corneal epithelia and as well as the regulation of both the barrier function of TJ and its ability to respond to EGF. Thus, TRPV4 function is necessary for the terminal differentiation of corneal epithelia.

  November 30, 2016   doi: 10.1002/jcp.25698   open full text
• S100B + A1 CELISA: A Novel Potency Assay and Screening Tool for Redifferentiation Stimuli of Human Articular Chondrocytes.
  Jose Diaz‐Romero, Sibylle Kürsener, Sandro Kohl, Dobrila Nesic.
  Journal of Cellular Physiology. November 30, 2016
  During monolayer expansion, a necessary step in autologous chondrocyte implantation, human articular chondrocytes (HAC) dedifferentiate and lose their capacity to produce stable hyaline cartilage. Determining HAC potency and learning how to trigger their redifferentiation would improve cell‐based cartilage regeneration therapies. We previously identified S100B and S100A1 proteins as markers of HAC redifferentiation potential. Here, we aimed to: (i) demonstrate a correlation between S100B + A1‐positive HAC in monolayer culture and their neochondrogenesis capacity in pellet culture; (ii) develop an S100B + A1 cell‐based ELISA, and (iii) prove that S100B + A1 induction in HAC increases their chondrogenic capacity. Expression patterns of S100A1 and S100B were investigated in HAC during dedifferentiation (monolayer) or redifferentiation (pellet or high‐osmolarity/BMP4 treatment in monolayer) using qRT‐PCR, immunocytochemistry, or immunohistochemistry. A cell‐based ELISA (CELISA) was developed as a 96‐well microplate multiplex assay to measure S100B + A1 (chondrogenesis), alkaline phosphatase (hypertrophy), and DNA amount (normalization), and applied to HAC, bone marrow‐derived mesenchymal stem cells and the chondrocytic cell line ATDC5. The direct correlation between the percentage of S100B + A1‐positive HAC in monolayer and their neochondrogenesis in pellets validates S100B + A1 as a marker of chondrogenic potency. The S100B + A1‐CELISA accurately determines HAC differentiation status, allows identification of chondrogenic stimuli, and permits the simultaneous monitoring of the undesirable hypertrophic phenotype. This novel assay offers a high‐throughput, comprehensive and versatile approach for measuring cell chondrogenic potency and for identifying redifferentiation factors/conditions. HAC improved neochondrogenesis in pellets—induced with high‐osmolarity and BMP4 treatment in monolayer—suggests that cell instruction prior to implantation may improve cartilage repair. J. Cell. Physiol. 232: 1559–1570, 2017. © 2016 Wiley Periodicals, Inc. The novel S100B + A1‐CELISA offers a high‐throughput, comprehensive and versatile approach for measuring cell chondrogenic potency and identifying redifferentiation factors/conditions to improve cartilage repair. Using a combination of high‐osmolarity medium and BMP4 treatment uncovered with this assay, we demonstrate that the induction of chondrocyte redifferentiation in monolayer improves their chondrogenic potential in pellets.

  November 30, 2016   doi: 10.1002/jcp.25682   open full text
• A Real‐World Multicentre Retrospective Study of Paclitaxel‐Bevacizumab and Maintenance Therapy as First‐Line for HER2‐Negative Metastatic Breast Cancer.
  Teresa Gamucci, Lucia Mentuccia, Clara Natoli, Isabella Sperduti, Alessandra Cassano, Andrea Michelotti, Luigi Di Lauro, Domenico Sergi, Alessandra Fabi, Maria G. Sarobba, Paolo Marchetti, Maddalena Barba, Emanuela Magnolfi, Marcello Maugeri‐Saccà, Ernesto Rossi, Valentina Sini, Antonino Grassadonia, Domenica Pellegrini, Antonino Astone, Cecilia Nisticò, Franco Angelini, Angela Vaccaro, Arianna Pellegrino, Claudia De Angelis, Michela Palleschi, Luca Moscetti, Ilaria Bertolini, Simonetta Buglioni, Antonio Giordano, Laura Pizzuti, Patrizia Vici.
  Journal of Cellular Physiology. November 30, 2016
  Bevacizumab in combination with taxanes in HER2‐negative metastatic breast cancer (MBC) patients has shown improved progression‐free survival (PFS), despite the lack of clear overall survival (OS) benefit. We performed a retrospective analysis to evaluate the impact of paclitaxel‐bevacizumab and of maintenance therapy with bevacizumab (BM) and endocrine therapy (ET) in the real‐world practice. We identified 314 HER2‐negative MBC patients treated in 12 cancer centers. Overall, the median PFS and OS were 14 and 40 months, respectively. Among the 254 patients potentially eligible for BM, 183 received BM after paclitaxel discontinuation until progression/toxicity. PFS and OS were improved in patients who had received BM in comparison with those potentially eligible but who did not receive BM (P< 0.0001 and P = 0.001, respectively). Results were confirmed when adjusting for propensity score. Among the 216 hormone‐receptor positive patients eligible for BM, a more favorable PFS and OS were observed when maintenance ET was administered (P < 0.0001). Multivariate analysis showed that PS, BM, number of disease sites and maintenance ET were related to PFS, while response and maintenance ET were related to OS. In hormone‐receptor positive patients, BM produced a significant PFS and a trend towards OS benefit only in absence of maintenance ET (P = 0.0007 and P = 0.06, respectively). In the triple‐negative subgroup, we observed a trend towards a better OS for patients who received BM (P = 0.06), without differences in PFS (P = 0.21). Our results confirmed the efficacy of first‐line paclitaxel‐bevacizumab in real‐world practice; both BM and maintenance ET significantly improved PFS and OS compared to no maintenance therapies. J. Cell. Physiol. 232: 1571–1578, 2017. © 2016 Wiley Periodicals, Inc. We performed a retrospective analysis to evaluate the impact of paclitaxel‐bevacizumab and of maintenance therapy with bevacizumab (BM) and endocrine therapy (ET) in the real‐world practice. Our results confirmed the efficacy of first‐line paclitaxel‐bevacizumab in real‐world practice; both BM and maintenance ET significantly improved PFS and OS compared to no maintenance therapies.

  November 30, 2016   doi: 10.1002/jcp.25685   open full text
• Oligodendrocyte Progenitor Cells Directly Utilize Lactate for Promoting Cell Cycling and Differentiation.
  Yoshinori Ichihara, Toru Doi, Youngjae Ryu, Motoshi Nagao, Yasuhiro Sawada, Toru Ogata.
  Journal of Cellular Physiology. November 30, 2016
  Oligodendrocyte progenitor cells (OPCs) undergo marked morphological changes to become mature oligodendrocytes, but the metabolic resources for this process have not been fully elucidated. Although lactate, a metabolic derivative of glycogen, has been reported to be consumed in oligodendrocytes as a metabolite, and to ameliorate hypomyelination induced by low glucose conditions, it is not clear about the direct contribution of lactate to cell cycling and differentiation of OPCs, and the source of lactate for remyelination. Therefore, we evaluated the effect of 1,4‐dideoxy‐1,4‐imino‐d‐arabinitol (DAB), an inhibitor of the glycogen catabolic enzyme glycogen phosphorylase, in a mouse cuprizone model. Cuprizone induced demyelination in the corpus callosum and remyelination occurred after cuprizone treatment ceased. This remyelination was inhibited by the administration of DAB. To further examine whether lactate affects proliferation or differentiation of OPCs, we cultured mouse primary OPC‐rich cells and analyzed the effect of lactate. Lactate rescued the slowed cell cycling induced by 0.4 mM glucose, as assessed by the BrdU‐positive cell ratio. Lactate also promoted OPC differentiation detected by monitoring the mature oligodendrocyte marker myelin basic protein, in the presence of both 36.6 mM and 0.4 mM glucose. Furthermore, these lactate‐mediated effects were suppressed by the reported monocarboxylate transporter inhibitor, α‐cyano‐4‐hydroxy‐cinnamate. These results suggest that lactate directly promotes the cell cycling rate and differentiation of OPCs, and that glycogen, one of the sources of lactate, contributes to remyelination in vivo. J. Cell. Physiol. 232: 986–995, 2017. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. Glycogen phosphorylase inhibitor suppresses remyelination in a cuprizone model. Lactate ameliorates low glucose‐induced slow cell–cell cycling and promotes differentiation of cell in OPC‐rich cultures. Monocarboxylase transport inhibition suppresses lactate effects in OPC‐rich cells.

  November 30, 2016   doi: 10.1002/jcp.25690   open full text
• CD93 as a Potential Target in Neovascular Age‐Related Macular Degeneration.
  Gian Marco Tosi, Elena Caldi, Barbara Parolini, Paolo Toti, Giovanni Neri, Federica Nardi, Claudio Traversi, Gabriele Cevenini, Davide Marigliani, Elisabetta Nuti, Tommaso Bacci, Federico Galvagni, Maurizio Orlandini.
  Journal of Cellular Physiology. November 30, 2016
  In patients with age‐related macular degeneration (AMD), choroidal neovascularization is the major cause of severe visual loss. In these patients, the persistence of neovascular growth despite vascular endothelial growth factor‐A blockage needs the discovery of new endothelial cell targets. The glycoprotein CD93, highly expressed in activated endothelial cells, has been recently involved in the regulation of the angiogenic process both as transmembrane and soluble protein. Choroidal neovascular membranes from patients affected by AMD were examined by immunofluorescence using anti‐CD93 and anti‐von Willebrand factor antibodies. Blood vessels within intraocular and extraocular neoplasias were used as controls for CD93 expression. All choroidal neovascular membranes displayed strong CD93 staining in the von Willebrand factor‐positive endothelial cells, consistently with the analyses showing a high colocalization coefficient in the blood vessels. Intraocular and extraocular tumor vessels showed similar results, whereas the normal choroid displayed blood vessels with only faint CD93 staining. Additionally, the concentration of soluble CD93 was determined in the aqueous humor of patients affected by naïve neovascular AMD by enzyme‐linked immunosorbent assays. Age‐matched cataract patients served as controls. Soluble CD93 was significantly increased in the aqueous humor of naïve neovascular AMD patients and tended to decrease after treatment with an antiangiogenic drug. In conclusion, both transmembrane and soluble CD93 are overexpressed in patients with neovascular AMD, indicating that CD93 may represent a potential new antiangiogenic target in the treatment of choroidal neovascularization. J. Cell. Physiol. 232: 1767–1773, 2017. © 2016 Wiley Periodicals, Inc. In patients with age‐related macular degeneration (AMD), the formation of new blood vessels in the choroid, which is the major cause of severe visual loss, persists even after therapy. In AMD patients, both in the endothelial cells of choroidal neovascular membranes and in the aqueous humor, we found high levels of the glycoprotein CD93, which has been recently involved in the enhancement of tumor vascular function. The presence of CD93 in AMD patients unveils a potential new target for choroidal neovascularization treatment.

  November 30, 2016   doi: 10.1002/jcp.25689   open full text
• E‐Cigarette Vapor Induces an Apoptotic Response in Human Gingival Epithelial Cells Through the Caspase‐3 Pathway.
  Mahmoud Rouabhia, Hyun Jin Park, Abdelhabib Semlali, Andrew Zakrzewski, Witold Chmielewski, Jamila Chakir.
  Journal of Cellular Physiology. November 30, 2016
  Electronic cigarettes represent an increasingly significant proportion of today's consumable tobacco products. E‐cigarettes contain several chemicals which may promote oral diseases. The aim of this study was to investigate the effect of e‐cigarette vapor on human gingival epithelial cells. Results show that e‐cigarette vapor altered the morphology of cells from small cuboidal form to large undefined shapes. Both single and multiple exposures to e‐cigarette vapor led to a bulky morphology with large faint nuclei and an enlarged cytoplasm. E‐cigarette vapor also increased L‐lactate dehydrogenase (LDH) activity in the targeted cells. This activity was greater with repeated exposures. Furthermore, e‐cigarette vapor increased apoptotic/necrotic epithelial cell percentages compared to that observed in the control. Epithelial cell apoptosis was confirmed by TUNEL assay showing that exposure to e‐cigarette vapor increased apoptotic cell numbers, particularly after two and three exposures. This negative effect involved the caspase‐3 pathway, the activity of which was greater with repeated exposure and which decreased following the use of caspase‐3 inhibitor. The adverse effects of e‐cigarette vapor on gingival epithelial cells may lead to dysregulated gingival cell function and result in oral disease. J. Cell. Physiol. 232: 1539–1547, 2017. © 2016 Wiley Periodicals, Inc. E‐cigarette vapor mediated gingival epithelial cell activity by promoting apoptosis which took place through an apoptosis pathway involving caspase‐3. The effects of e‐cigarette vapor on gingival epithelial cells may compromise epithelial tissue, resulting in periodontal disease development and potentiating inflammation resulting from the tissue damage.

  November 30, 2016   doi: 10.1002/jcp.25677   open full text
• Low Shear Stress Attenuates COX‐2 Expression Induced by Resistin in Human Osteoarthritic Chondrocytes.
  Yu‐Ping Su, Cheng‐Nan Chen, Hsin‐I Chang, Kuo‐Chin Huang, Chin‐Chang Cheng, Fang‐Yao Chiu, Ko‐Chao Lee, Chun‐Min Lo, Shun‐Fu Chang.
  Journal of Cellular Physiology. November 20, 2016
  Low shear stress has been proposed to play a reparative role in modulating cartilage homeostasis. Recently, epidemiological studies have found a positive correlation between the resistin level in serum and synovial fluid and osteoarthritis (OA) severity in patients. However, the effect of moderate shear stress on the catabolic stimulation of resistin in OA chondrocytes remains unclear. Hence, this study was to investigate whether low shear stress could regulate resistin‐induced catabolic cyclooxygenase (COX)‐2 expression in human OA chondrocytes and the underlying mechanism. Human OA chondrocytes and SW1353 chondrosarcoma cells were used in this study. Two modes of low shear stress (2 dyn/cm2), pre‐shear and post‐shear, were applied to the chondrocytes. A specific activator and siRNAs were used to investigate the mechanism of low shear stress‐regulated COX‐2 expression of resistin induction. We found that human OA chondrocytes exposed to different modes of low shear stress elicit an opposite effect on resistin‐induced COX‐2 expression: pre‐shear for a short duration attenuates the resistin effect by inhibiting the transcription factor nuclear factor (NF)‐κB‐p65 subunit and the cAMP response element binding protein; however, post‐shear over a longer duration enhances the resistin effect by activating only the NF‐κB‐p65 subunit. Moreover, our results demonstrated that the regulation of both shear modes in resistin‐stimulated COX‐2 expression occurs through increasing AMP‐activated protein kinase activation and then sirtuin 1 expression. This study elucidates the detailed mechanism of low shear stress regulating the resistin‐induced catabolic COX‐2 expression and indicates a possible reparative role of moderate shear force in resistin‐stimulated OA development. J. Cell. Physiol. 232: 1448–1457, 2017. © 2016 Wiley Periodicals, Inc. This study elucidates the detailed mechanism of low shear stress regulating the resistin‐induced catabolic COX‐2 expression and indicates a possible reparative role of moderate shear force in resistin‐stimulated OA development.

  November 20, 2016   doi: 10.1002/jcp.25644   open full text
• Calcium Signaling During Meiotic Cell Cycle Regulation and Apoptosis in Mammalian Oocytes.
  Meenakshi Tiwari, Shilpa Prasad, Tulsidas G. Shrivastav, Shail K. Chaube.
  Journal of Cellular Physiology. November 20, 2016
  Calcium (Ca++) is one of the major signal molecules that regulate various aspects of cell functions including cell cycle progression, arrest, and apoptosis in wide variety of cells. This review summarizes current knowledge on the differential roles of Ca++ in meiotic cell cycle resumption, arrest, and apoptosis in mammalian oocytes. Release of Ca++ from internal stores and/or Ca++ influx from extracellular medium causes moderate increase of intracellular Ca++ ([Ca++]i) level and reactive oxygen species (ROS). Increase of Ca++ as well as ROS levels under physiological range trigger maturation promoting factor (MPF) destabilization, thereby meiotic resumption from diplotene as well as metaphase‐II (M‐II) arrest in oocytes. A sustained increase of [Ca++]i level beyond physiological range induces generation of ROS sufficient enough to cause oxidative stress (OS) in aging oocytes. The increased [Ca++]i triggers Fas ligand‐mediated oocyte apoptosis. Further, OS triggers mitochondria‐mediated oocyte apoptosis in several mammalian species. Thus, Ca++ exerts differential roles on oocyte physiology depending upon its intracellular concentration. A moderate increase of [Ca++]i as well as ROS mediate spontaneous resumption of meiosis from diplotene as well as M‐II arrest, while their high levels cause meiotic cell cycle arrest and apoptosis by operating both mitochondria‐ as well as Fas ligand‐mediated apoptotic pathways. Indeed, Ca++ regulates cellular physiology by modulating meiotic cell cycle and apoptosis in mammalian oocytes. J. Cell. Physiol. 232: 976–981, 2017. © 2016 Wiley Periodicals, Inc. A moderate increase of [Ca++]i level under physiological range induces generation of ROS that result in decrease of cAMP level and increase of CaMKII activity. A decrease of intraoocyte cAMP level and increase of CaMKII activity directly and/or indirectly triggers MPF destabilization thereby meiotic resumption from diplotene as well as M‐II arrest. The high level of [Ca++]i beyond physiological range triggers Fas ligand‐mediated apoptosis and induces OS. The increased OS triggers oocyte apoptosis through mitochondria‐mediated pathway. Indeed, high level of [Ca++]i and ROS‐mediated meiotic cell cycle arrest and apoptosis could be interlinked in mammalian oocytes.

  November 20, 2016   doi: 10.1002/jcp.25670   open full text
• Notch Signaling Pathway Regulates Angiogenesis via Endothelial Cell in 3D Co‐Culture Model.
  Dan Zhao, Changyue Xue, Shiyu Lin, Sirong Shi, Qianshun Li, Mengting Liu, Xiaoxiao Cai, Yunfeng Lin.
  Journal of Cellular Physiology. November 20, 2016
  This study aimed to investigate the role of Notch signaling pathway for angiogenesis in a three‐dimensional (3D) collagen gel model with co‐culture of adipose‐derived stromal cells (ASCs) and endothelial cells (ECs). A 3D collagen gel model was established in vitro by implanting both ASCs from green fluorescent protein‐labeled mouse and ECs from red fluorescent protein‐labeled mouse, and the phenomena of angiogenesis with Notch signaling inducer Jagged1, inhibitor DAPT and PBS, respectively were observed by confocal laser scanning microscopy. Semi‐quantitative PCR and immunofluorescent staining were conducted to detect expressions of angiogenesis‐related genes and proteins. Angiogenesis in the co‐culture gels was promoted by Jagged1 treatment while attenuated by DAPT treatment, compared to control group. In co‐culture system of ASCs and ECs, the gene expressions of VEGFA, VEGFB, Notch1, Notch2, Hes1, Hey1, VEGFR1,and the protein expression of VEGFA, VEGFB, Notch1, Hes1, Hey1 were increased by Jagged1 treatment and decreased by DAPT treatment in ECs. And the result of VEGFR3 was the opposite. However, the same results did not appear completely in ASCs. These results revealed the VEGFA/B‐Notch1/2‐Hes1/Hey1‐ VEGFR1/3 signal axis played an important role in angiogenesis when ASCs and ECs were co‐cultured in a 3D collagen gel model. J. Cell. Physiol. 232: 1548–1558, 2017. © 2016 Wiley Periodicals, Inc. VEGFA/B‐Notch1/2‐Hes1/Hey1‐ VEGFR1/3 signal axis played an important role in angiogenesis when ASCs and ECs were co‐cultured in a 3D collagen gel model.

  November 20, 2016   doi: 10.1002/jcp.25681   open full text
• Wnt3A Induces GSK‐3β Phosphorylation and β‐Catenin Accumulation Through RhoA/ROCK.
  Jae‐Gyu Kim, Myoung‐Ju Kim, Won‐Ji Choi, Mi‐Young Moon, Hee‐Jun Kim, Jae‐Yong Lee, Jaebong Kim, Sung‐Chan Kim, Seung Goo Kang, Goo‐Young Seo, Pyeung‐Hyeun Kim, Jae‐Bong Park.
  Journal of Cellular Physiology. November 20, 2016
  In canonical pathway, Wnt3A has been known to stabilize β‐catenin through the dissociation between β‐catenin and glycogen synthase kinase‐3β (GSK‐3β) that suppresses the phosphorylation and degradation of β‐catenin. In non‐canonical signaling pathway, Wnt was known to activate Rho GTPases and to induce cell migration. The cross‐talk between canonical and non‐canonical pathways by Wnt signaling; however, has not been fully elucidated. Here, we revealed that Wnt3A induces not only the phosphorylation of GSK‐3β and accumulation of β‐catenin but also RhoA activation in RAW264.7 and HEK293 cells. Notably, sh‐RhoA and Tat‐C3 abolished both the phosphorylation of GSK‐3β and accumulation of β‐catenin. Y27632, an inhibitor of Rho‐associated coiled coil kinase (ROCK) and si‐ROCK inhibited both GSK‐3β phosphorylation and β‐catenin accumulation. Furthermore, active domain of ROCK directly phosphorylated the purified recombinant GSK‐3β in vitro. In addition, Wnt3A‐induced cell proliferation and migration, which were inhibited by Tat‐C3 and Y27632. Taken together, we propose the cross‐talk between canonical and non‐canonical signaling pathways of Wnt3A, which induces GSK‐3β phosphorylation and β‐catenin accumulation through RhoA and ROCK activation. J. Cell. Physiol. 232: 1104–1113, 2017. © 2016 Wiley Periodicals, Inc. Wnt3A activates RhoA and ROCK, which induces GSK‐3beta phosphorylation and beta‐catenin accumulation. RhoA and ROCK are involved in the regulation of cell proliferation and migration upon Wnt3A.

  November 20, 2016   doi: 10.1002/jcp.25572   open full text
• ADP‐Induced Ca2+ Signaling and Proliferation of Rat Ventricular Myofibroblasts Depend on Phospholipase C‐Linked TRP Channels Activation Within Lipid Rafts.
  Mariana Certal, Adriana Vinhas, Aurora Barros‐Barbosa, Fátima Ferreirinha, Maria Adelina Costa, Paulo Correia‐de‐Sá.
  Journal of Cellular Physiology. November 10, 2016
  Nucleotides released during heart injury affect myocardium electrophysiology and remodeling through P2 purinoceptors activation in cardiac myofibroblasts. ATP and UTP endorse [Ca2+]i accumulation and growth of DDR‐2/α‐SMA‐expressing myofibroblasts from adult rat ventricles via P2Y4 and P2Y2 receptors activation, respectively. Ventricular myofibroblasts also express ADP‐sensitive P2Y1, P2Y12, and P2Y13 receptors as demonstrated by immunofluorescence confocal microscopy and western blot analysis, but little information exists on ADP effects in these cells. ADP (0.003–3 mM) and its stable analogue, ADPßS (100 μM), caused fast [Ca2+]i transients originated from thapsigargin‐sensitive internal stores, which partially declined to a plateau sustained by capacitative Ca2+ entry through transient receptor potential (TRP) channels inhibited by 2‐APB (50 μM) and flufenamic acid (100 μM). Hydrophobic interactions between Gq/11‐coupled P2Y purinoceptors and TRP channels were suggested by prevention of the ADP‐induced [Ca2+]i plateau following PIP2 depletion with LiCl (10 mM) and cholesterol removal from lipid rafts with methyl‐ß‐cyclodextrin (2 mM). ADP [Ca2+]i transients were insensitive to P2Y1, P2Y12, and P2Y13 receptor antagonists, MRS2179 (10μM), AR‐C66096 (0.1 μM), and MRS2211 (10μM), respectively, but were attenuated by suramin and reactive blue‐2 (100 μM) which also blocked P2Y4 receptors activation by UTP. Cardiac myofibroblasts growth and type I collagen production were favored upon activation of MRS2179‐sensitive P2Y1 receptors with ADP or ADPßS (30 μM). In conclusion, ADP exerts a dual role on ventricular myofibroblasts: [Ca2+]i transients are mediated by fast‐desensitizing P2Y4 receptors, whereas the pro‐fibrotic effect of ADP involves the P2Y1 receptor activation. Data also show that ADP‐induced capacitative Ca2+ influx depends on phospholipase C‐linked TRP channels opening in lipid raft microdomains. J. Cell. Physiol. 232: 1511–1526, 2017. © 2016 Wiley Periodicals, Inc. The P2Y4R mainly mediates ADP‐induced [Ca2+]i rise in rat ventricular myofibroblasts; ADP‐induced fast [Ca2+]i rises depend on Ca2+ recruitment from internal stores; the late [Ca2+]i plateau depends on capacitative Ca2+ entry via TRP channels; PLC‐activating P2Y4 receptors and TRP channels are assembled in lipid rafts; ADP promotes cell growth and collagen production predominantly via P2Y1 activation.

  November 10, 2016   doi: 10.1002/jcp.25656   open full text
• Peripheral Blood Mononuclear Cells Infiltration Downregulates Decidual FAAH Activity in an LPS‐Induced Embryo Resorption Model.
  Manuel Luis Wolfson, Julieta Aisemberg, Fernando Correa, Ana María Franchi.
  Journal of Cellular Physiology. November 10, 2016
  Maternal infections with gram‐negative bacteria are associated with miscarriage and are one of the most common complications during pregnancy. Previous studies from our group have shown that lipopolysaccharide (LPS)‐activated infiltrating peripheral blood mononuclear cells (PBMC) into decidual tissue plays an important role in the establishment of a local inflammatory process that results in embryo cytotoxicity and early embryo resorption. Moreover, we have also shown that an increased endocannabinoid tone mediates LPS‐induced deleterious effects during early pregnancy loss. Here, we sought to investigate whether the infiltrating PBMC modulates the decidual endocannabinoid tone and the molecular mechanisms involved. PBMC isolated from 7‐day pregnant mice subjected to different treatments were co‐cultured in a transwell system with decidual tissue from control 7‐day pregnant mice. Decidual fatty acid amide hydrolase (FAAH) activity was measured by radioconvertion, total decidual protein nitration by Western blot (WB), and decidual FAAH nitration by immunoprecipitation followed by WB. We found that co‐culture of PBMC obtained from LPS‐treated mice increased the level of nitration of decidual FAAH, which resulted in a negative modulation of decidual FAAH activity. Interestingly, co‐treatment with progesterone or aminoguanidine prevented this effect. We found that LPS‐treated PBMC release high amounts of nitric oxide (NO) which causes tyrosine nitration of decidual FAAH, diminishing its enzymatic activity. Inactivation of FAAH, the main degrading enzyme of anandamide and similar endocannabinoids, could lead to an increased decidual endocannabinoid tone with embryotoxic effects. J. Cell. Physiol. 232: 1441–1447, 2017. © 2016 Wiley Periodicals, Inc. PBMC release a soluble factor which modulates the decidual FAAH activity. Peroxynitrite participates in LPS‐treated PBMC decidual FAAH activity modulation. PBMC from LPS‐treated mice increased the nitration levels of decidual FAAH which resulted in a decreased enzymatic activity.

  November 10, 2016   doi: 10.1002/jcp.25640   open full text
• p120‐Catenin Is Required for Dietary Calcium Suppression of Oral Carcinogenesis in Mice.
  Zhongjian Xie, Yuan Yuan, Yi Jiang, Chandrama Shrestha, Ying Chen, Liyan Liao, Shangli Ji, Xiaoge Deng, Eryuan Liao, Daniel D. Bikle.
  Journal of Cellular Physiology. November 10, 2016
  Previous studies have shown that dietary calcium suppresses oral carcinogenesis, but the mechanism is unclear. p120‐catenin (p120) is a cytoplasmic protein closely associated with E‐cadherin to form the E‐cadherin–β‐catenin complex and may function as a tumor suppressor in the oral epithelium. To determine whether p120 is involved in the mechanism by which dietary calcium suppresses oral carcinogenesis, The normal, low, or high calcium diet was fed control mice (designated as floxed p120 mice) or mice in which p120 was specifically deleted in the oral squamous epithelium during the adult stage (designated as p120cKO mice). All mice were exposed to a low dose of oral cancer carcinogen 4‐nitroquinoline 1‐oxide and rates of oral squamous cell carcinoma (OSCC) and proliferation and differentiation in the cancerous and non‐cancerous oral epithelium of these mice were examined. The results showed that the low calcium diet increased rates of OSCC and proliferation of the non‐cancerous oral epithelium and decreased differentiation of the non‐cancerous oral epithelium, but had no effect on cancerous oral epithelium. In contrast, the high calcium diet had opposite effects. However, the effect of the dietary calcium on the rates of OSCC, proliferation, and differentiation of the non‐cancerous epithelium were not seen in p120cKO mice. Based on these results, we conclude that p120 is required for dietary calcium suppression of oral carcinogenesis and oral epithelial proliferation and dietary calcium induction of oral epithelial differentiation. J. Cell. Physiol. 232: 1360–1367, 2017. © 2016 Wiley Periodicals, Inc. The aim of the present study was to determine whether p120‐catenin plays a role in dietary calcium‐induced oral carcinogenesis. The results showed that knockout of p120‐catenin blocked the inhibitory effect of dietary calcium on tumor rates and proliferation and the stimulatory effect of dietary calcium on tumor differentiation in the oral epithelium. These data suggest that p120‐catenin plays a critical role in dietary calcium‐suppressed carcinogenesis and dietary calcium regulation of proliferation, differentiation. Our findings illustrate a mechanism by which dietary calcium suppresses oral carcinogenesis will help in the development of better preventive and therapeutic strategies for oral cancers.

  November 10, 2016   doi: 10.1002/jcp.25620   open full text
• Functional Roles of Eph A‐Ephrin A1 System in Endometrial Luminal Epithelial Cells During Early Pregnancy.
  Whasun Lim, Hyocheol Bae, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. November 10, 2016
  Eph and ephrin regulate diverse biological events such as proliferation, adhesion, migration, and angiogenesis through cell‐to‐cell interactions. However, little is known of their functional role and mechanisms of action in uterine endometrial cells. In the present study, we demonstrated the effects of the Eph and ephrin on interactions between blastocysts and endometrial luminal epithelial (pLE) cells in the pig that is regarded as an excellent biomedical animal model for research on the peri‐implantation period of pregnancy. Results of this study indicated that among eight members of the Eph A family, expression of Eph A1, A2, A4, and A7 was strongly detected in endometrial epithelial cells during early pregnancy. Of these, for identification of signal transduction pathways induced by ephrin A1, a major ligand for Eph A, cell proliferation assays, and immunofluorescence and cell cycle regulation were analyzed following treatment of pLE cells with ephrin A1. Ephrin A1 stimulated proliferation of pLE cells as evidenced by abundant PCNA expression and an increase in the G2/M phase. Western blot analysis showed that ephrin A1 activated PI3K and MAPK signaling proteins in a time‐dependent manner. Moreover, phosphorylation of AKT, ERK1/2, P38, and JNK proteins were suppressed by their inhibitors wortmannin, U0126, SB203580, and SP600125, respectively. Also, phosphor‐AKT was reduced by ERK1/2 and P38 inhibitors. Ephrin A1‐induced proliferation and migration of pLE cells was also blocked by those inhibitors. Collectively, these results suggest that ephrin A1 enhances interactions between porcine blastocysts and endometrial luminal epithelial cells by activating PI3K and MAPK signal transduction pathways. J. Cell. Physiol. 232: 1527–1538, 2017. © 2016 Wiley Periodicals, Inc. Eph A‐ephrin A1 system plays an important role in uterine receptivity and improvement of litter size through PI3K/AKT and MAPK signal transduction pathways during the peri‐implantation period of pregnancy.

  November 10, 2016   doi: 10.1002/jcp.25659   open full text
• Extracellular Matrix and Colorectal Cancer: How Surrounding Microenvironment Affects Cancer Cell Behavior?
  Sara Crotti, Martina Piccoli, Flavio Rizzolio, Antonio Giordano, Donato Nitti, Marco Agostini.
  Journal of Cellular Physiology. November 10, 2016
  Colorectal cancer (CRC) whit more than a million of new cases per year is one of the most common registered cancers worldwide with few treatment options especially for advanced and metastatic patients.The tumor microenvironment is composed by extracellular matrix (ECM), cells, and interstitial fluids. Among all these constituents, in the last years an increased interest around the ECM and its potential role in cancer tumorigenesis is arisen. During cancer progression the ECM structure and composition became disorganized, allowing cellular transformation and metastasis. Up to now, the focus has mainly been on the characterization of CRC microenvironment analyzing separately structural ECM components or cell secretome modifications. A more extensive view that interconnects these aspects should be addressed. In this review, biochemical (secretome) and biomechanical (structure and architecture) changes of tumor microenvironment will be discussed, giving suggestions on how these changes can affect cancer cell behavior. J. Cell. Physiol. 232: 967–975, 2017. © 2016 Wiley Periodicals, Inc. In this review we discuss how changes in one of the major components of tumoral niche, the extracellular matrix (ECM), allow disease initiation, progression and aggressiveness. The ECM remodeling process in colorectal cancer is presented here considering the alterations occurring in the architecture/structure and in the secretome composition, giving suggestions of new interesting markers of aggressiveness. A comprehensive view of the interactions between tumor cells and ECM should be further investigate in the future to provide new and helpful markers of patients’ response to therapy.

  November 10, 2016   doi: 10.1002/jcp.25658   open full text
• Valproic Acid and Lithium Meditate Anti‐Inflammatory Effects by Differentially Modulating Dendritic Cell Differentiation and Function.
  Sy‐Jye Leu, Yi‐Yuan Yang, Hsing‐Cheng Liu, Chieh‐Yu Cheng, Yu‐Chen Wu, Ming‐Chyi Huang, Yuen‐Lun Lee, Chi‐Ching Chen, Winston W. Shen, Ko‐Jiunn Liu.
  Journal of Cellular Physiology. November 10, 2016
  Valproic acid (VPA), with inhibition activity mainly toward histone deacetylase (HDAC) and Glycogen Synthase Kinase (GSK)‐3, and lithium, with inhibition activity mainly toward GSK‐3, are both prescribed in clinical as mood‐stabilizers and anticonvulsants for the control of bipolar disorder. This study aims to compare the immuno‐modulation activities of VPA and lithium, especially on the differentiation and functions of dendritic cells (DC). Our data show that treatment with VPA or lithium effectively alleviated the severity of collagen‐induced arthritis triggered by LPS in mice. Both agents reduced the serum level of IL‐6 and IL‐10 after LPS challenge in mice. VPA and lithium both induce significant down‐regulation of group I CD1 expression and secretion of IL‐6 during differentiation of human monocyte‐derived immature DC, while they differ in the induction of CD83 and CD86 expression, secretion of IL‐8, IL‐10, and TNF‐α. Upon stimulation of immature DC with LPS, VPA, and lithium both reduced the secretion of IL‐6 and TNF‐α. However, only lithium significantly increased the production of IL‐10, while VPA increased the production of IL‐8 but substantially reduce the secretion of IL‐10 and IL‐23. Treatment with VPA resulted in a reduced capacity of LPS‐stimulated DC to promote the differentiation of T helper 17 cells that are critical in the promotion of inflammatory responses. Taken together, our results suggest that VPA and lithium may differentially modulate inflammation through regulating the capacity of DC to mediate distinct T cell responses, and they may provide a complementary immunomodulatory effects for the treatment of inflammation‐related diseases. J. Cell. Physiol. 232: 1176–1186, 2017. © 2016 Wiley Periodicals, Inc. Upon stimulation of immature DC with LPS, VPA, and lithium both reduced the secretion of IL‐6 and TNF‐α. However, only lithium significantly increased the production of IL‐10, while VPA increased the production of IL‐8 but substantially reduce the secretion of IL‐10 and IL‐23. Treatment with VPA resulted in a reduced capacity of LPS‐stimulated DC to promote the differentiation of T helper 17 cells. VPA and lithium may differentially modulate inflammation through regulating the capacity of DC to mediate distinct T cell responses, and they may provide a complementary immunomodulatory effects for the treatment of inflammation‐related diseases.

  November 10, 2016   doi: 10.1002/jcp.25604   open full text
• Src Family Kinase Links Insulin Signaling to Short Term Regulation of Na,K‐ATPase in Nonpigmented Ciliary Epithelium.
  Mohammad Shahidullah, Amritlal Mandal, Nicholas A. Delamere.
  Journal of Cellular Physiology. November 10, 2016
  Insulin has been shown to elicit changes of Na,K‐ATPase activity in various tissues. Na,K‐ATPase in the nonpigmented ciliary epithelium (NPE) plays a role in aqueous humor secretion and changes of Na,K‐ATPase activity impact the driving force. Because we detect a change of NPE Na,K‐ATPase activity in response to insulin, studies were carried out to examine the response mechanism. Ouabain‐sensitive rubidium (Rb) uptake by cultured NPE cells, measured as a functional index of Na,K‐ATPase‐mediated inward potassium transport, was found to increase in cells exposed for 5 min to insulin. The maximally effective concentration was 100 nM. An intrinsic increase of Na,K‐ATPase activity evident as a >2‐fold increase in the rate of ouabain‐sensitive ATP hydrolysis in homogenates obtained from cells exposed to 100 nM insulin for 5 min was also observed. Insulin‐treated cells exhibited Akt, Src family kinase (SFK), ERK1/2, and p38 activation, all of which were prevented by a pI3 kinase inhibitor LY294002. The Rb uptake and Na,K‐ATPase activity response to insulin both were abolished by PP2, an SFK inhibitor which also prevented p38 and ERK1/2 but not Akt activation. The Akt inhibitor MK‐2206 did not change the Na,K‐ATPase response to insulin. The findings suggest insulin activates pI3K‐dependent Akt and SFK signaling pathways that are separate. ERK1/2 and p38 activation is secondary to and dependent on SFK activation. The increase of Na,K‐ATPase activity is dependent on activation of the SFK pathway. The findings are consistent with previous studies that indicate a link between Na,K‐ATPase activity and SFK signaling. J. Cell. Physiol. 232: 1489–1500, 2017. © 2016 Wiley Periodicals, Inc. Studies were conducted to examine the insulin response in ocular nonpigmented ciliary epithelium. Insulin elicited a pronounced short term increase in Na,K‐ATPase activity and a complex signaling response that involved Akt (protein kinase B), Src family kinase (SFK), ERK1/2, and p38 MAPK all of which were dependent on PI3K activation. ERK1/2 and p38 MAPK activation were secondary to and dependent on SFK activation. The insulin‐dependent increase in Na,K‐ATPase activity was linked to SFK activation. In contrast, Akt activation has no discernable effect on Na,K‐ATPase activity.

  November 10, 2016   doi: 10.1002/jcp.25654   open full text
• Neuroprotection and Blood–Brain Barrier Restoration by Salubrinal After a Cortical Stab Injury.
  M. Asunción Barreda‐Manso, Natalia Yanguas‐Casás, Manuel Nieto‐Sampedro, Lorenzo Romero‐Ramírez.
  Journal of Cellular Physiology. November 10, 2016
  Following a central nervous system (CNS) injury, restoration of the blood‐brain barrier (BBB) integrity is essential for recovering homeostasis. When this process is delayed or impeded, blood substances and cells enter the CNS parenchyma, initiating an additional inflammatory process that extends the initial injury and causes so‐called secondary neuronal loss. Astrocytes and profibrotic mesenchymal cells react to the injury and migrate to the lesion site, creating a new glia limitans that restores the BBB. This process is beneficial for the resolution of the inflammation, neuronal survival, and the initiation of the healing process. Salubrinal is a small molecule with neuroprotective properties in different animal models of stroke and trauma to the CNS. Here, we show that salubrinal increased neuronal survival in the neighbourhood of a cerebral cortex stab injury. Moreover, salubrinal reduced cortical blood leakage into the parenchyma of injured animals compared with injured controls. Adjacent to the site of injury, salubrinal induced immunoreactivity for platelet‐derived growth factor subunit B (PDGF‐B), a specific mitogenic factor for mesenchymal cells. This effect might be responsible for the increased immunoreactivity for fibronectin and the decreased activation of microglia and macrophages in injured mice treated with salubrinal, compared with injured controls. The immunoreactivity for PDGF‐B colocalized with neuronal nuclei (NeuN), suggesting that cortical neurons in the proximity of the injury were the main source of PDGF‐B. Our results suggest that after an injury, neurons play an important role in both, the healing process and the restoration of the BBB integrity. J. Cell. Physiol. 232: 1501–1510, 2017. © 2016 Wiley Periodicals, Inc. Salubrinal is neuroprotective in an animal model of cortical stab injury and facilitated the restoration of the BBB integrity, by inducing the neuronal production of PDGF‐B. Our results suggest that neurons in the proximity of the lesion have a previously underrated, major role in the restoration of the BBB.

  November 10, 2016   doi: 10.1002/jcp.25655   open full text
• The Emerging Role of TRAF7 in Tumor Development.
  Tiziana Zotti, Ivan Scudiero, Pasquale Vito, Romania Stilo.
  Journal of Cellular Physiology. November 03, 2016
  The seven members of the tumor necrosis factor receptor (TNF‐R)‐associated factor (TRAF) family of intracellular proteins were originally discovered and characterized as signaling adaptor molecules coupled to the cytoplasmic regions of receptors of the TNF‐R superfamily. Functionally, TRAFs act both as a scaffold and/or enzymatic proteins to regulate activation of mitogen‐activated protein kinases (MAPKs) and transcription factors of nuclear factor‐κB family (NF‐κB). Given the wide variety of stimuli intracellularly conveyed by TRAF proteins, they are physiologically involved in multiple biological processes, including embryonic development, tissue homeostasis and regulation of innate and adaptive immune responses. In the last few years, it has become increasingly evident the involvement of TRAF7, the last member of the TRAF family to be discovered, in the genesis and progression of several human cancers, placing TRAF7 in the spotlight as a novel tumor suppressor protein. In this paper, we review and discuss the literature recently produced on this subject. This article is protected by copyright. All rights reserved

  November 03, 2016   doi: 10.1002/jcp.25676   open full text
• Novel Instruments for the Implementation of Electrochemotherapy Protocols: From Bench Side to Veterinary Clinic.
  Enrico P. Spugnini, Stefano Fais, Tommaso Azzarito, Alfonso Baldi.
  Journal of Cellular Physiology. October 27, 2016
  Electrochemotherapy (ECT) is a medical strategy that allows an increased efficacy of chemotherapy agents after the application of permeabilizing electric pulses having appropriate characteristics (form, voltage, frequency). In the past 10 years, the clinical efficacy of this therapeutic approach in several spontaneous models of tumors in animals has been shown. Moreover, some of the molecular and cellular mechanisms responsible for this phenomenon have been elucidated. Our group has been deeply involved in the development of new ECT protocols for companion animals, implementing the use of the technique as first line treatment, and evaluating different chemotherapy agents in laboratory animals as well as pets. This article summarizes the most important advances in veterinary ECT, including the development of novel equipment, therapeutic protocols, and their translation to humans. J. Cell. Physiol. 232: 490–495, 2017. © 2016 Wiley Periodicals, Inc. Electrochemotherapy (ECT) is a medical strategy that allows an increased efficacy of chemotherapy agents after the application of permeabilizing electric pulses having appropriate characteristics (form, voltage, frequency). Our group has been deeply involved in the development of new ECT protocols for companion animals, implementing the use of the technique as first line treatment, and evaluating different chemotherapy agents in laboratory animals as well as pets. This article summarizes the most important advances in veterinary ECT, including the development of novel equipment, therapeutic protocols, and their translation to humans.

  October 27, 2016   doi: 10.1002/jcp.25505   open full text
• Pathophysiology of the Desmo‐Adhesome.
  Antonio Celentano, Michele Davide Mignogna, Michael McCullough, Nicola Cirillo.
  Journal of Cellular Physiology. October 27, 2016
  Advances in our understanding of desmosomal diseases have provided a clear demonstration of the key role played by desmosomes in tissue and organ physiology, highlighting the importance of their dynamic and finely regulated structure. In this context, non‐desmosomal regulatory molecules have acquired increasing relevance in the study of this organelle resulting in extending the desmosomal interactome, named the “desmo‐adhesome.” Spatiotemporal changes in the expression and regulation of the desmo‐adhesome underlie a number of genetic, infectious, autoimmune, and malignant conditions. The aim of the present article was to examine the structural and functional relationship of the desmosome, by providing a comprehensive, yet focused overview of the constituents targeted in human disease. The inclusion of the novel regulatory network in the desmo‐adhesome pathophysiology opens new avenues to a deeper understanding of desmosomal diseases, potentially unveiling pathogenic mechanisms waiting to be explored. J. Cell. Physiol. 232: 496–505, 2017. © 2016 Wiley Periodicals, Inc. The desmosome has been the subject of intense research over the last 20 years. While several reviews have recently been published on this topic, none has investigated the desmosomal interactome as a whole. This review is unique in that it not only takes into account the diseases that are linked to primary desmosomal components but, also, includes the potential role of accessory molecules in epithelial disease and cancer.

  October 27, 2016   doi: 10.1002/jcp.25515   open full text
• Role of microRNAs on the Regulation of Mitochondrial Biogenesis and Insulin Signaling in Skeletal Muscle.
  Tanes I. Lima, Hygor N. Araujo, Eveline S. Menezes, Carlos H. Sponton, Michel B. Araújo, Lucas H.M. Bomfim, André L. Queiroz, Madla A. Passos, Thais Amaral e Sousa, Sandro M. Hirabara, Amanda R. Martins, Helena C.L.B. Sampaio, Alice Rodrigues, Rui Curi, Everardo M. Carneiro, Antônio C. Boschero, Leonardo R. Silveira.
  Journal of Cellular Physiology. October 26, 2016
  Mitochondria play a critical role in several cellular processes and cellular homeostasis. Mitochondrion dysfunction has been correlated with numerous metabolic diseases such as obesity and type 2 diabetes. MicroRNAs are non‐coding RNAs that have emerged as key regulators of cell metabolism. The microRNAs act as central regulators of metabolic gene networks by leading to the degradation of their target messenger RNA or repression of protein translation. In addition, vesicular and non‐vesicular circulating miRNAs exhibit a potential role as mediators of the cross‐talk between the skeletal muscle and other tissues/organs. In this review, we will focus on the emerging knowledge of miRNAs controlling mitochondrial function and insulin signaling in skeletal muscle cells. J. Cell. Physiol. 232: 958–966, 2017. © 2016 Wiley Periodicals, Inc. Mitochondrion dysfunction in skeletal muscle has been correlated with innumerous metabolic diseases. MicroRNAs have emerged as key regulators of cell metabolism. In addition, vesicular and non‐vesicular circulating miRNAs exhibit potential role as mediators of the cross‐talking between skeletal muscle and other tissues controlling mitochondrial function and insulin signaling.

  October 26, 2016   doi: 10.1002/jcp.25645   open full text
• Increased Chondrogenic Potential of Mesenchymal Cells From Adipose Tissue Versus Bone Marrow‐Derived Cells in Osteoarthritic In Vitro Models.
  Stefania Pagani, Veronica Borsari, Francesca Veronesi, Andrea Ferrari, Simona Cepollaro, Paola Torricelli, Giuseppe Filardo, Milena Fini.
  Journal of Cellular Physiology. October 26, 2016
  Primarily, to compare the behavior of human mesenchymal stem cells (MSCs) derived from bone marrow (hBMSCs) and adipose tissue (hADSCs) in an osteoarthritic (OA) microenvironment; secondly, to investigate the reaction of these cell types in two alternative in vitro culture systems, obtained by using TNFα and/or IL1β as inflammation mediators, or by using synovial fluid harvested by OA patients (OSF) to simulate the complex inflamed knee microenvironment. 3D micromass cultures of hBMSCs or hADSCs were grown in chondrogenic medium (CTR), in the presence of TNFα and/or IL1β, or synovial fluid from OA patients. After 1 month of culture, the chondrogenic differentiation of micromasses was evaluated by gene expression, matrix composition, and organization. Both hMSCs types formed mature micromasses in CTR, but a better response of hADSCs to the inflammatory environment was documented by micromass area and Bern score evaluations. The addition of OSF elicited a milder reaction than with TNFα and/or IL1β by both cell types, probably due to the presence of both catabolic and protective factors. In particular, SOX9 and ACAN gene expression and GAG synthesis were more abundant in hADSCs than hBMSCs when cultured in OSF. The expression of MMP1 was increased for both hMSCs in inflammatory conditions, but in particular by hBMSCs. hADSCs showed an increased chondrogenic potential in inflammatory culture systems, suggesting a better response of hADSCs in the OA environment, thus underlining the importance of appropriate in vitro models to study MSCs and potential advantages of using these cells for future clinical applications. J. Cell. Physiol. 232: 1478–1488, 2017. © 2016 Wiley Periodicals, Inc. Primary objective of this paper was to compare the behavior of human mesenchymal stem cells (MSCs) derived from bone marrow (hBMSCs) and adipose tissue (hADSCs) in an osteoarthritic (OA) microenvironment; secondly, to investigate the reaction of these cell types in two alternative in vitro culture systems, obtained by using TNFα and/or IL1β as inflammation mediators, or by using synovial fluid harvested by OA patients (OSF) to simulate the complex inflamed knee microenvironment.

  October 26, 2016   doi: 10.1002/jcp.25651   open full text
• Essential Role of CFTR in PKA‐Dependent Phosphorylation, Alkalinization, and Hyperpolarization During Human Sperm Capacitation.
  Lis C. Puga Molina, Nicolás A. Pinto, Paulina Torres Rodríguez, Ana Romarowski, Alberto Vicens Sanchez, Pablo E. Visconti, Alberto Darszon, Claudia L. Treviño, Mariano G. Buffone.
  Journal of Cellular Physiology. October 26, 2016
  Mammalian sperm require to spend a limited period of time in the female reproductive tract to become competent to fertilize in a process called capacitation. It is well established that HCO3− is essential for capacitation because it activates the atypical soluble adenylate cyclase ADCY10 leading to cAMP production, and promotes alkalinization of cytoplasm, and membrane hyperpolarization. However, how HCO3− is transported into the sperm is not well understood. There is evidence that CFTR activity is involved in the human sperm capacitation but how this channel is integrated in the complex signaling cascades associated with this process remains largely unknown. In the present work, we have analyzed the extent to which CFTR regulates different events in human sperm capacitation. We observed that inhibition of CFTR affects HCO3−‐entrance dependent events resulting in lower PKA activity. CFTR inhibition also affected cAMP/PKA‐downstream events such as the increase in tyrosine phosphorylation, hyperactivated motility, and acrosome reaction. In addition, we demonstrated for the first time, that CFTR and PKA activity are essential for the regulation of intracellular pH, and membrane potential in human sperm. Addition of permeable cAMP partially recovered all the PKA‐dependent events altered in the presence of inh‐172 which is consistent with a role of CFTR upstream of PKA activation. J. Cell. Physiol. 232: 1404–1414, 2017. © 2016 Wiley Periodicals, Inc. We observed that inhibition of CFTR affects HCO3‐entrance dependent events resulting in lower PKA activity.

  October 26, 2016   doi: 10.1002/jcp.25634   open full text
• Interfering of the Reelin/ApoER2/PSD95 Signaling Axis Reactivates Dendritogenesis of Mature Hippocampal Neurons.
  Estibaliz Ampuero, Nur Jury, Steffen Härtel, María‐Paz Marzolo, Brigitte van Zundert.
  Journal of Cellular Physiology. October 25, 2016
  Reelin, an extracellular glycoprotein secreted in embryonic and adult brain, participates in neuronal migration and neuronal plasticity. Extensive evidence shows that reelin via activation of the ApoER2 and VLDLR receptors promotes dendrite and spine formation during early development. Further evidence suggests that reelin signaling is needed to maintain a stable architecture in mature neurons, but, direct evidence is lacking. During activity‐dependent maturation of the neuronal circuitry, the synaptic protein PSD95 is inserted into the postsynaptic membrane to induce structural refinement and stability of spines and dendrites. Given that ApoER2 interacts with PSD95, we tested if reelin signaling interference in adult neurons reactivates the dendritic architecture. Unlike findings in developing cultures, the presently obtained in vitro and in vivo data show, for the first time, that reelin signaling interference robustly increase dendritogenesis and reduce spine density in mature hippocampal neurons. In particular, the expression of a mutant ApoER2 form (ApoER2‐tailless), which is unable to interact with PSD95 and hence cannot transduce reelin signaling, resulted in robust dendritogenesis in mature hippocampal neurons in vitro. These results indicate that reelin/ApoER2/PSD95 signaling is important for neuronal structure maintenance in mature neurons. Mechanistically, obtained immunofluorescent data indicate that reelin signaling impairment reduced synaptic PSD95 levels, consequently leading to synaptic re‐insertion of NR2B‐NMDARs. Our findings underscore the importance of reelin in maintaining adult network stability and reveal a new mode for reactivating dendritogenesis in neurological disorders where dendritic arbor complexity is limited, such as in depression, Alzheimer's disease, and stroke. J. Cell. Physiol. 232: 1187–1199, 2017. © 2016 Wiley Periodicals, Inc. In mature neurons the reelin interference reactivates the dendritogenesis in the adults reelin signaling maintains adult network stability.

  October 25, 2016   doi: 10.1002/jcp.25605   open full text
• Heat Stress Modulates Both Anabolic and Catabolic Signaling Pathways Preventing Dexamethasone‐Induced Muscle Atrophy In Vitro.
  Wakako Tsuchida, Masahiro Iwata, Takayuki Akimoto, Shingo Matsuo, Yuji Asai, Shigeyuki Suzuki.
  Journal of Cellular Physiology. October 19, 2016
  It is generally recognized that synthetic glucocorticoids induce skeletal muscle weakness, and endogenous glucocorticoid levels increase in patients with muscle atrophy. It is reported that heat stress attenuates glucocorticoid‐induced muscle atrophy; however, the mechanisms involved are unknown. Therefore, we examined the mechanisms underlying the effects of heat stress against glucocorticoid‐induced muscle atrophy using C2C12 myotubes in vitro, focusing on expression of key molecules and signaling pathways involved in regulating protein synthesis and degradation. The synthetic glucocorticoid dexamethasone decreased myotube diameter and protein content, and heat stress prevented the morphological and biochemical glucocorticoid effects. Heat stress also attenuated increases in mRNAs of regulated in development and DNA damage responses 1 (REDD1) and Kruppel‐like factor 15 (KLF15). Heat stress recovered the dexamethasone‐induced inhibition of PI3K/Akt signaling. These data suggest that changes in anabolic and catabolic signals are involved in heat stress‐induced protection against glucocorticoid‐induced muscle atrophy. These results have a potentially broad clinical impact because elevated glucocorticoid levels are implicated in a wide range of diseases associated with muscle wasting. J. Cell. Physiol. 232: 650–664, 2017. © 2016 The Authors. Journal of Cellular Physiology published by Wiley Periodicals, Inc. Heat stress attenuated increases in REDD1 and KLF15, two genes directly targeted by glucocorticoids that decrease protein synthesis by inhibiting mTORC1. Heat stress also normalized dexamethasone‐induced increases in key factors promoting protein degradation through the ubiquitin‐proteasome system, including KLF15.

  October 19, 2016   doi: 10.1002/jcp.25609   open full text
• Short‐ and Long‐Term Hindlimb Immobilization and Reloading: Profile of Epigenetic Events in Gastrocnemius.
  Alba Chacon‐Cabrera, Joaquim Gea, Esther Barreiro.
  Journal of Cellular Physiology. October 19, 2016
  Skeletal muscle dysfunction and atrophy are characteristic features accompanying chronic conditions. Epigenetic events regulate muscle mass and function maintenance. We hypothesized that the pattern of epigenetic events (muscle‐enriched microRNAs and histone acetylation) and acetylation of transcription factors known to signal muscle wasting may differ between early‐ and late‐time points in skeletal muscles of mice exposed to hindlimb immobilization (I) and recovery following I. Body and muscle weights, grip strength, muscle‐enriched microRNAs, histone deacetylases (HDACs), acetylation of proteins, histones, and transcription factors (TF), myogenic TF factors, and muscle phenotype were assessed in gastrocnemius of mice exposed to periods (1, 2, 3, 7, 15, and 30 days, I groups) of hindlimb immobilization, and in those exposed to reloading for different periods of time (1, 3, 7, 15, and 30 days, R groups) following 7‐day immobilization. Compared to non‐immobilized controls, muscle weight, limb strength, microRNAs, especially miR‐486, SIRT1 levels, and slow‐ and fast‐twitch cross‐sectional areas were decreased in mice of I groups, whereas Pax7 and acetylated FoxO1 and FoxO3 levels were increased. Muscle reloading following splint removal improved muscle mass loss, strength, and fiber atrophy, by increasing microRNAs, particularly miR‐486, and SIRT1 content, while decreasing acetylated FoxO1 and FoxO3 levels. In this mouse model of disuse muscle atrophy, muscle‐enriched microRNAs, especially miR‐486, through Pax7 regulation delayed muscle cell differentiation following unloading of gastrocnemius muscle. Acetylation of FoxO1 and 3 seemed to drive muscle mass loss and atrophy, while deacetylation of these factors through SIRT1 would enable the muscle fibers to regenerate. J. Cell. Physiol. 232: 1415–1427, 2017. © 2016 Wiley Periodicals, Inc. In this mouse model of disuse muscle atrophy, muscle‐enriched microRNAs, especially miR‐486, through Pax7 regulation delayed muscle cell differentiation following unloading of gastrocnemius muscle. Acetylation of FoxO1 and 3 seemed to drive muscle mass loss and atrophy, while deacetylation of these factors through SIRT1 would enable the muscle fibers to regenerate.

  October 19, 2016   doi: 10.1002/jcp.25635   open full text
• Osteoclast Fusion: Time‐Lapse Reveals Involvement of CD47 and Syncytin‐1 at Different Stages of Nuclearity.
  Anaïs Marie Julie Møller, Jean‐Marie Delaissé, Kent Søe.
  Journal of Cellular Physiology. October 19, 2016
  Investigations addressing the molecular keys of osteoclast fusion are primarily based on end‐point analyses. No matter if investigations are performed in vivo or in vitro the impact of a given factor is predominantly analyzed by counting the number of multi‐nucleated cells, the number of nuclei per multinucleated cell or TRAcP activity. But end‐point analyses do not show how the fusion came about. This would not be a problem if fusion of osteoclasts was a random process and occurred by the same molecular mechanism from beginning to end. However, we and others have in the recent period published data suggesting that fusion partners may specifically select each other and that heterogeneity between the partners seems to play a role. Therefore, we set out to directly test the hypothesis that fusion factors have a heterogenic involvement at different stages of nuclearity. Therefore, we have analyzed individual fusion events using time‐lapse and antagonists of CD47 and syncytin‐1. All time‐lapse recordings have been studied by two independent observers. A total of 1808 fusion events were analyzed. The present study shows that CD47 and syncytin‐1 have different roles in osteoclast fusion depending on the nuclearity of fusion partners. While CD47 promotes cell fusions involving mono‐nucleated pre‐osteoclasts, syncytin‐1 promotes fusion of two multi‐nucleated osteoclasts, but also reduces the number of fusions between mono‐nucleated pre‐osteoclasts. Furthermore, CD47 seems to mediate fusion mostly through broad contact surfaces between the partners’ cell membrane while syncytin‐1 mediate fusion through phagocytic‐cup like structure. J. Cell. Physiol. 232: 1396–1403, 2017. © 2016 Wiley Periodicals, Inc. Time‐lapse recording has allowed us to categorize cell fusion events according to the number of nuclei of the fusion partners. We show here that factors commonly believed to be critical for fusion, are in fact only involved in some categories of fusion. This points to the existence of distinct molecular mechanisms of cell fusion depending on the degree of nuclearity.

  October 19, 2016   doi: 10.1002/jcp.25633   open full text
• MicroRNA‐494 Activation Suppresses Bone Marrow Stromal Cell‐Mediated Drug Resistance in Acute Myeloid Leukemia Cells.
  Chen Tian, Guoguang Zheng, Hongqing Zhuang, Xubin Li, Dongzhi Hu, Lei Zhu, Tengteng Wang, Mingjian James You, Yizhuo Zhang.
  Journal of Cellular Physiology. October 19, 2016
  Acute myeloid leukemia (AML) is not sensitive to chemotherapy partially because of the protection of AML cells by mesenchymal stromal cells (MSCs). Our previous studies found that MSCs protected AML cells from apoptosis through the c‐Myc‐dependent pathway. However, the mechanism by which MSCs regulate c‐Myc in AML cells is still unknown. To elucidate the mechanism, we performed microRNA array analysis of AML cell lines and validated by TaqMan realtime PCR. The results showed that the expression of microRNA‐494 (miR‐494) in AML cells after coculture with MSCs was downregulated. Reporter gene analysis confirmed miR‐494 as one of the regulators of c‐Myc. In the coculture system, activation of miR‐494 in AML cells suppressed proliferation and induced apoptosis of AML cells in vitro. After addition of mitoxantrone to the coculture system, the proliferation of AML cells with miR‐494 activation was suppressed more than that of control cells. After subcutaneous injection of AML cell lines in combination with MSC, tumor growth was suppressed in mice injected with miR‐494‐overexpressing AML cells. The rate of tumor formation was even lower after mitoxantrone treatment in the miR‐494 overexpressing group. Moreover, miR‐494 activation resulted in a decrease of leukemic cell counts in peripheral blood (PB) and bone marrow, and prolonged survival in mice injected with miR‐494‐overexpressing AML cellls and MSCs compared to the control mice. Our results indicate that miR‐494 suppresses drug resistance in AML cells by downregulating c‐Myc through interaction with MSCs and that miR‐494 therefore is a potential therapeutic target. J. Cell. Physiol. 232: 1387–1395, 2017. © 2016 Wiley Periodicals, Inc. miR‐494 suppresses drug resistance in AML cells by downregulating c‐Myc through interaction with MSCs and that miR‐494 therefore is a potential therapeutic target.

  October 19, 2016   doi: 10.1002/jcp.25628   open full text
• Increased Amyloid Precursor Protein and Tau Expression Manifests as Key Secondary Cell Death in Chronic Traumatic Brain Injury.
  Sandra A. Acosta, Naoki Tajiri, Paul R. Sanberg, Yuji Kaneko, Cesar V. Borlongan.
  Journal of Cellular Physiology. October 19, 2016
  In testing the hypothesis of Alzheimer's disease (AD)‐like pathology in late stage traumatic brain injury (TBI), we evaluated AD pathological markers in late stage TBI model. Sprague–Dawley male rats were subjected to moderate controlled cortical impact (CCI) injury, and 6 months later euthanized and brain tissues harvested. Results from H&E staining revealed significant 33% and 10% reduction in the ipsilateral and contralateral hippocampal CA3 interneurons, increased MHCII‐activated inflammatory cells in many gray matter (8–20‐fold increase) and white matter (6–30‐fold increased) regions of both the ipsilateral and contralateral hemispheres, decreased cell cycle regulating protein marker by 1.6‐ and 1‐fold in the SVZ and a 2.3‐ and 1.5‐fold reductions in the ipsilateral and contralateral dentate gyrus, diminution of immature neuronal marker by two‐ and onefold in both the ipsilateral and contralateral SVZ and dentate gyrus, and amplified amyloid precursor protein (APP) distribution volumes in white matter including corpus callosum, fornix, and internal capsule (4–38‐fold increase), as well as in the cortical gray matter, such as the striatum hilus, SVZ, and dentate gyrus (6–40‐fold increase) in TBI animals compared to controls (P's < 0.001). Surrogate AD‐like phenotypic markers revealed a significant accumulation of phosphorylated tau (AT8) and oligomeric tau (T22) within the neuronal cell bodies in ipsilateral and contralateral cortex, and dentate gyrus relative to sham control, further supporting the rampant neurodegenerative pathology in TBI secondary cell death. These findings indicate that AD‐like pathological features may prove to be valuable markers and therapeutic targets for late stage TBI. J. Cell. Physiol. 232: 665–677, 2017. © 2016 Wiley Periodicals, Inc. In testing the hypothesis of Alzheimer's disease (AD)‐like pathology in late stage traumatic brain injury (TBI), we evaluated AD pathological markers in late stage TBI model. We provided evidence that neuronal cell loss, inflammation, decreased cell proliferation and impaired neurogenesis, occurred in discreet neurostructures exhibiting abnormal intra‐neuronal APP expressions, indicating an extensive AD‐like neuropathology in advanced TBI. Moreover, surrogate AD‐like phenotypic markers revealed a significant accumulation of phosphorylated tau (AT8) and oligomeric tau (T22) within the neuronal cell bodies in ipsilateral and contralateral cortex, and dentate gyrus relative to sham control, further supporting the rampant neurodegenerative pathology in TBI secondary cell death. These findings indicate that AD‐like pathological features may prove to be valuable markers and therapeutic targets for late stage TBI.

  October 19, 2016   doi: 10.1002/jcp.25629   open full text
• Indispensable Role of Ion Channels and Transporters in the Auditory System.
  Rahul Mittal, Mayank Aranke, Luca H. Debs, Desiree Nguyen, Amit P. Patel, M'hamed Grati, Jeenu Mittal, Denise Yan, Prem Chapagain, Adrien A. Eshraghi, Xue Zhong Liu.
  Journal of Cellular Physiology. October 17, 2016
  Ear is a complex system where appropriate ionic composition is essential for maintaining the tissue homeostasis and hearing function. Ion transporters and channels present in the auditory system plays a crucial role in maintaining proper ionic composition in the ear. The extracellular fluid, called endolymph, found in the cochlea of the mammalian inner ear is particularly unique due to its electrochemical properties. At an endocochlear potential of about +80 mV, signaling initiated by acoustic stimuli at the level of the hair cells is dependent on the unusually high potassium (K+) concentration of endolymph. There are ion channels and transporters that exists in the ear to ensure that K+ is continually being cycled into the stria media endolymph. This review is focused on the discussion of the molecular and genetic basis of previously and newly recognized ion channels and transporters that support sensory hair cell excitation based on recent knock‐in and knock‐out studies of these channels. This article also addresses the molecular and genetic defects and the pathophysiology behind Meniere's disease as well as how the dysregulation of these ion transporters can result in severe defects in hearing or even deafness. Understanding the role of ion channels and transporters in the auditory system will facilitate in designing effective treatment modalities against ear disorders including Meniere's disease and hearing loss. J. Cell. Physiol. 232: 743–758, 2017. © 2016 Wiley Periodicals, Inc. We review the role of ion channels and transporters in the auditory system and summarize recent advancements in understanding the role of ion channels using genetic and molecular tools. Dysregulation of ion channels and transporters causes ear disorders including Meniere's disease and hearing loss. SLC22 family of ion transporters plays a crucial role in maintaining the integrity of stria vascularis and preventing age‐related hearing loss.

  October 17, 2016   doi: 10.1002/jcp.25631   open full text
• Early Growth Response 3 (Egr3) Contributes a Maintenance of C2C12 Myoblast Proliferation.
  Mitsutoshi Kurosaka, Yuji Ogura, Toshiya Funabashi, Tatsuo Akema.
  Journal of Cellular Physiology. October 12, 2016
  Satellite cell proliferation is a crucially important process for adult myogenesis. However, its regulatory mechanisms remain unknown. Early growth response 3 (Egr3) is a zinc‐finger transcription factor that regulates different cellular functions. Reportedly, Egr3 interacts with multiple signaling molecules that are also known to regulate satellite cell proliferation. Therefore, it is possible that Egr3 is involved in satellite cell proliferation. Results of this study have demonstrated that Egr3 transcript levels are upregulated in regenerating mouse skeletal muscle after cardiotoxin injury. Using C2C12 myoblast culture (a model of activated satellite cells), results show that inhibition of Egr3 by shRNA impairs the myoblast proliferation rate. Results also show reduction of NF‐кB transcriptional activity in Egr3‐inhibited cells. Inhibition of Egr3 is associated with an increase in annexin V+ cell fraction and apoptotic protein activity including caspase‐3 and caspase‐7, and Poly‐ADP ribose polymerase. By contrast, the reduction of cellular proliferation by inhibition of Egr3 was partially recovered by treatment of pan‐caspase inhibitor Z‐VAD‐FMK. Collectively, these results suggest that Egr3 is involved in myoblast proliferation by interaction with survival signaling. J. Cell. Physiol. 232: 1114–1122, 2017. © 2016 Wiley Periodicals, Inc. Egr3 maintains myoblast proliferation by stabilizing basal NF‐κB activity. However, the inhibition of Egr3 results in activation of apoptotic signaling by suppressing NF‐κB activity. The activation of apoptotic signaling is associated with proliferation defect in myoblast.

  October 12, 2016   doi: 10.1002/jcp.25574   open full text
• Gene and MicroRNA Expression Are Predictive of Tumor Response in Rectal Adenocarcinoma Patients Treated With Preoperative Chemoradiotherapy.
  Caterina Millino, Isacco Maretto, Beniamina Pacchioni, Maura Digito, Antonino De Paoli, Vincenzo Canzonieri, Edoardo D'Angelo, Marco Agostini, Flavio Rizzolio, Antonio Giordano, Andrea Barina, Senthilkumar Rajendran, Giovanni Esposito, Gerolamo Lanfranchi, Donato Nitti, Salvatore Pucciarelli.
  Journal of Cellular Physiology. October 07, 2016
  Preoperative chemoradiotherapy (pCRT) followed by surgery is the standard treatment for locally advanced rectal cancer (LARC). However, tumor response to pCRT is not uniform, and there are no effective predictive methods. This study investigated whether specific gene and miRNA expression are associated with tumor response to pCRT. Tissue biopsies were obtained from patients before pCRT and resection. Gene and miRNA expression were analyzed using a one‐color microarray technique that compares signatures between responders (R) and non‐responders (NR), as measured based on tumor regression grade. Two groups composed of 38 “exploration cohort” and 21 “validation cohort” LARC patients were considered for a total of 32 NR and 27 R patients. In the first cohort, using SAM Two Class analysis, 256 genes and 29 miRNAs that were differentially expressed between the NR and R patients were identified. The anti‐correlation analysis showed that the same 8 miRNA interacted with different networks of transcripts. The miR‐630 appeared only with the NR patients and was anti‐correlated with a single transcript: RAB5B. After PAM, the following eight transcripts were strong predictors of tumor response: TMEM188, ITGA2, NRG, TRAM1, BCL2L13, MYO1B, KLF7, and GTSE1. Using this gene set, an unsupervised cluster analysis was applied to the validation cohort and correctly assigned the patients to the NR or R group with 85.7% accuracy, 90% sensitivity, and 82% specificity. All three parameters reached 100% when both cohorts were considered together. In conclusion, gene and miRNA expression profiles may be helpful for predicting response to pCRT in LARC patients. J. Cell. Physiol. 232: 426–435, 2017. © 2016 Wiley Periodicals, Inc. Preoperative chemoradiotherapy (pCRT) followed by surgery is the standard treatment for locally advanced rectal cancer. This study investigated whether specific gene and miRNA expression are associated with tumor response to pCRT.

  October 07, 2016   doi: 10.1002/jcp.25441   open full text
• Pulsed Electromagnetic Field Exposure Reduces Hypoxia and Inflammation Damage in Neuron‐Like and Microglial Cells.
  Fabrizio Vincenzi, Annalisa Ravani, Silvia Pasquini, Stefania Merighi, Stefania Gessi, Stefania Setti, Ruggero Cadossi, Pier Andrea Borea, Katia Varani.
  Journal of Cellular Physiology. October 07, 2016
  In the present study, the effect of low‐frequency, low‐energy pulsed electromagnetic fields (PEMFs) has been investigated by using different cell lines derived from neuron‐like cells and microglial cells. In particular, the primary aim was to evaluate the effect of PEMF exposure in inflammation‐ and hypoxia‐induced injury in two different neuronal cell models, the human neuroblastoma‐derived SH‐SY5Y cells and rat pheochromocytoma PC12 cells and in N9 microglial cells. In neuron‐like cells, live/dead and apoptosis assays were performed in hypoxia conditions from 2 to 48 h. Interestingly, PEMF exposure counteracted hypoxia damage significantly reducing cell death and apoptosis. In the same cell lines, PEMFs inhibited the activation of the hypoxia‐inducible factor 1α (HIF‐1α), the master transcriptional regulator of cellular response to hypoxia. The effect of PEMF exposure on reactive oxygen species (ROS) production in both neuron‐like and microglial cells was investigated considering their key role in ischemic injury. PEMFs significantly decreased hypoxia‐induced ROS generation in PC12, SH‐SY5Y, and N9 cells after 24 or 48 h of incubation. Moreover, PEMFs were able to reduce some of the most well‐known pro‐inflammatory cytokines such as tumor necrosis factor–α (TNF‐α), interleukin (IL)‐1β, IL‐6, and IL‐8 release in N9 microglial cells stimulated with different concentrations of LPS for 24 or 48 h of incubation time. These results show a protective effect of PEMFs on hypoxia damage in neuron‐like cells and an anti‐inflammatory effect in microglial cells suggesting that PEMFs could represent a potential therapeutic approach in cerebral ischemic conditions. J. Cell. Physiol. 232: 1200–1208, 2017. © 2016 Wiley Periodicals, Inc. In the present study, the effect of low‐frequency, low‐energy pulsed electromagnetic fields (PEMFs) has been investigated by using different cell lines derived from neuron‐like cells and microglial cells. PEMF exposure counteracted hypoxia damage significantly reducing cell death and apoptosis, reduced HIF‐1α, ROS, and proinflammatory cytokines. PEMFs could represent a potential therapeutic approach in cerebral ischemic conditions.

  October 07, 2016   doi: 10.1002/jcp.25606   open full text
• Inflammatory Events and Oxidant Production in the Diaphragm, Gastrocnemius, and Blood of Rats Exposed to Chronic Intermittent Hypoxia: Therapeutic Strategies.
  Marisol Domínguez‐Álvarez, Joaquim Gea, Esther Barreiro.
  Journal of Cellular Physiology. September 30, 2016
  We hypothesized that inflammatory events and reactive oxygen species (ROS) production may be differentially expressed in respiratory and limb muscles, and blood of a chronic intermittent hypoxia (CIH) experimental model and that antioxidants and TNF‐alpha blockade may influence those events. In blood, diaphragm, and gastrocnemius of rats non‐invasively exposed to CIH (10% hypoxia, 2 h/day, 14 consecutive days) with/without concomitant treatment with either anti‐TNF‐alpha antibody (infliximab) or N‐acetyl cysteine (NAC), inflammatory cytokines, superoxide anion production, muscle structural abnormalities, and fiber‐type composition were assessed. Compared to non‐exposed controls, in CIH‐exposed rats, body weight gain was reduced, TNF‐alpha, IL‐1beta, IL‐6, and interferon‐gamma levels were increased in diaphragm, TNF‐alpha, and IL‐1 beta plasma levels were greater, systemic and muscle superoxide anion production was higher, diaphragm and gastrocnemius inflammatory cells and internal nuclei were higher, and muscle fiber‐type and morphometry remained unmodified. CIH rats treated with infliximab further increased TNF‐alpha, IL‐1beta, IL‐6, and interferon‐gamma diaphragm levels, whereas NAC induced a reduction only in TNF‐alpha and IL‐1beta levels in diaphragm and plasma. Infliximab and NAC elicited a significant decline in superoxide anion production in diaphragm, gastrocnemius, and plasma, while inducing a further increase in inflammatory cells and internal nuclei in both muscles. Proinflammatory cytokines are differentially expressed in respiratory and limb muscles and plasma of CIH‐exposed rats, while superoxide anion production increased in both muscle types and blood. Infliximab and NAC exerted different effects. These findings may help understand the biology underlying CIH in skeletal muscles and blood of patients with chronic respiratory diseases. J. Cell. Physiol. 232: 1165–1175, 2017. © 2016 Wiley Periodicals, Inc. Proinflammatory cytokines are differentially expressed in respiratory and limb muscles and plasma of CIH‐exposed rats, while superoxide anion production increased in both muscle types and blood. Infliximab and NAC exerted different effects. These findings may help understand the biology underlying CIH in skeletal muscles and blood of patients with chronic respiratory diseases.

  September 30, 2016   doi: 10.1002/jcp.25600   open full text
• MED28 Regulates Epithelial–Mesenchymal Transition Through NFκB in Human Breast Cancer Cells.
  Chun‐Yin Huang, Nien‐Tsu Hsieh, Chun‐I Li, Yu‐Ting Weng, Hsiao‐Sheng Liu, Ming‐Fen Lee.
  Journal of Cellular Physiology. September 30, 2016
  MED28, a mammalian Mediator subunit, was found highly expressed in several types of malignancy, including breast cancer. Recently, we have identified a role of MED28 in regulating both cell growth and migration in human breast cancer cells. In epithelium‐derived solid tumor, migration and invasion are preceded by the progression of epithelial–mesenchymal transition (EMT) which calls for downregulation of epithelial markers as well as upregulation of mesenchymal markers, among other features. The objective of this study was to investigate a putative role of MED28 in the progression of EMT in human breast cancer cells. In fibroblast‐like MDA‐MB‐231 cells, suppression of MED28 attenuated the mesenchymal morphology, concomitantly with a reduction of several mesenchymal biomarkers and Snail, a transcriptional repressor of E‐cadherin. The suppression effect was also accompanied by downregulation of p‐NFκB/p65. However, overexpression of MED28 exhibited in an opposite manner. In epithelial MCF7 cells, administration of Adriamycin®, an experimental EMT induction system, led to a mesenchyme‐like appearance correlated with increased expression of MED28, p‐p65, and Snail, and a reciprocal change of epithelial and mesenchymal markers. Furthermore, suppression of MED28 attenuated the experimental EMT effect and restored the original expression status of E‐cadherin and MMP9 in MCF7 cells. Our data indicate that MED28 modulates the development of EMT through NFκB in human breast cancer cells, further reinforcing the significance of MED28 in the progression of breast cancer on top of its role in cell growth and migration. J. Cell. Physiol. 232: 1337–1345, 2017. © 2016 Wiley Periodicals, Inc. MED28 regulates the expression of epithelial markers and mesenchymal markers through NFκB, thereby modulating the development of epithelial–mesenchymal transition. These findings further reinforce the significance of MED28 in the progression of breast cancer on top of its role in cell growth and migration.

  September 30, 2016   doi: 10.1002/jcp.25610   open full text
• Hypoglycemia Enhances Epithelial‐Mesenchymal Transition and Invasiveness, and Restrains the Warburg Phenotype, in Hypoxic HeLa Cell Cultures and Microspheroids.
  Álvaro Marín‐Hernández, Juan Carlos Gallardo‐Pérez, Ileana Hernández‐Reséndiz, Isis Del Mazo‐Monsalvo, Diana Xochiquetzal Robledo‐Cadena, Rafael Moreno‐Sánchez, Sara Rodríguez‐Enríquez.
  Journal of Cellular Physiology. September 30, 2016
  The accelerated growth of solid tumors leads to episodes of both hypoxia and hypoglycemia (HH) affecting their intermediary metabolism, signal transduction, and transcriptional activity. A previous study showed that normoxia (20% O2) plus 24 h hypoglycemia (2.5 mM glucose) increased glycolytic flux whereas oxidative phosphorylation (OxPhos) was unchanged versus normoglycemia in HeLa cells. However, the simultaneous effect of HH on energy metabolism has not been yet examined. Therefore, the effect of hypoxia (0.1–1% O2) plus hypoglycemia on the energy metabolism of HeLa cells was analyzed by evaluating protein content and activity, along with fluxes of both glycolysis and OxPhos. Under hypoxia, in which cell growth ceased and OxPhos enzyme activities, ΔΨm and flux were depressed, hypoglycemia did not stimulate glycolytic flux despite increasing H‐RAS, p‐AMPK, GLUT1, GLUT3, and HKI levels, and further decreasing mitochondrial enzyme content. The impaired mitochondrial function in HH cells correlated with mitophagy activation. The depressed OxPhos and unchanged glycolysis pattern was also observed in quiescent cells from mature multicellular tumor spheroids, suggesting that these inner cell layers are similarly subjected to HH. The principal ATP supplier was glycolysis for HH 2D monolayer and 3D quiescent spheroid cells. Accordingly, the glycolytic inhibitors iodoacetate and gossypol were more effective than mitochondrial inhibitors in decreasing HH‐cancer cell viability. Under HH, stem cell‐, angiogenic‐, and EMT‐biomarkers, as well as glycoprotein‐P content and invasiveness, were also enhanced. These observations indicate that HH cancer cells develop an attenuated Warburg and pronounced EMT‐ and invasive‐phenotype. J. Cell. Physiol. 232: 1346–1359, 2017. © 2016 Wiley Periodicals, Inc. Hypoglycemia enhances (i) epithelial‐mesenchymal transition, (ii) invasiveness, and (iii) restrains the Warburg phenotype in hypoxic HeLa cells.

  September 30, 2016   doi: 10.1002/jcp.25617   open full text
• PACAP and VIP Inhibit HIF‐1α‐Mediated VEGF Expression in a Model of Diabetic Macular Edema.
  Grazia Maugeri, Agata Grazia D'Amico, Salvatore Saccone, Concetta Federico, Sebastiano Cavallaro, Velia D'Agata.
  Journal of Cellular Physiology. September 30, 2016
  Pituitary adenylate cyclase‐activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) exert a protective role against retinal injuries, including diabetic macular edema (DME). The macular damage is induced by hyperglycemia, which damages vessels supplying blood to the retina and induces hypoxia. The microenvironmental changes stimulate the expression of hypoxia‐inducible factors (HIFs), which promote the choroidal endothelial cell transmigration across the retinal pigmented epithelium (RPE) into neurosensory retina, where they proliferate into new vessels under stimulation of the vascular endothelial growth factor (VEGF). In the present study, we have investigated whether PACAP and VIP prevent retinal damage by modulating the expression of HIFs, VEGF, and its receptors. In accord to our hypothesis, we have shown that both peptides are able to significantly reduce HIF‐1α and increase HIF‐3α expression in ARPE‐19 cells exposed to hyperglycemic/hypoxic insult. This effect is also related to a reduction of VEGF and its receptors expression. Moreover, both peptides also reduce the activation of p38 mitogen‐activated protein kinase (MAPK), a pro‐apoptotic signaling pathway, which is activated by VEGFR‐1 and 2 receptors. In conclusion, our study has further elucidated the protective role performed by PACAP and VIP, against the harmful combined effect of hyperglycemia/hypoxia characterizing the DME microenvironment. J. Cell. Physiol. 232: 1209–1215, 2017. © 2016 Wiley Periodicals, Inc. PACAP and VIP prevent retinal damage by modulating the expression of HIFs, VEGF, and its receptors; PACAP and VIP act against the harmful combined effect of hyperglycemia/hypoxia.

  September 30, 2016   doi: 10.1002/jcp.25616   open full text
• Differences in Activation of HIV‐1 Replication by Superinfection With HIV‐1 and HIV‐2 in U1 Cells.
  Xue Wang, Bing Sun, Christelle Mbondji, Santanu Biswas, Jiangqin Zhao, Indira Hewlett.
  Journal of Cellular Physiology. September 30, 2016
  Macrophages contribute to HIV‐1 pathogenesis by forming a viral reservoir that serve as a viral source for the infection of CD4 T cells. The relationship between HIV‐1 latent infection and superinfection in macrophages has not been well studied. Using susceptible U1 cells chronically infected with HIV‐1, we studied the effects of HIV superinfection on latency and differences in superinfection with HIV‐1 and HIV‐2 in macrophages. We found that HIV‐1 (MN) superinfection displayed increased HIV‐1 replication in a time‐dependent manner; while cells infected with HIV‐2 (Rod) initially showed increased HIV‐1 replication, followed by a decrease in HIV‐1 RNA production. HIV‐1 superinfection upregulated/activated NF‐ĸB, NFAT, AP‐1, SP‐1, and MAPK Erk through expression/activation of molecules, CD4, CD3, TCRβ, Zap‐70, PLCγ1, and PKCΘ in T cell receptor‐related signaling pathways; while HIV‐2 superinfection initially increased expression/activation of these molecules followed by decreased protein expression/activation. HIV superinfection initially downregulated HDAC1 and upregulated acetyl‐histone H3 and histone H3 (K4), while HIV‐2 superinfection demonstrated an increase in HDAC1 and a decrease in acetyl‐histone H3 and histone H3 (K4) relative to HIV‐1 superinfection. U1 cells superinfected with HIV‐1 or HIV‐2 showed differential expression of proteins, IL‐2, PARP‐1, YB‐1, and LysRS. These findings indicate that superinfection with HIV‐1 or HIV‐2 has different effects on reactivation of HIV‐1 replication. HIV‐1 superinfection with high load of viral replication may result in high levels of cytotoxicity relative to HIV‐2 superinfection. Cells infected with HIV‐2 showed lower level of HIV‐1 replication, suggesting that co‐infection with HIV‐2 may result in slower progression toward AIDS. J. Cell. Physiol. 232: 1746–1753, 2017. © 2016 Wiley Periodicals, Inc.

  September 30, 2016   doi: 10.1002/jcp.25614   open full text
• Risk Differences Between Prediabetes And Diabetes According To Breast Cancer Molecular Subtypes.
  A. Crispo, L.S.A. Augustin, M. Grimaldi, F. Nocerino, A. Giudice, E. Cavalcanti, M. Di Bonito, G. Botti, M. De Laurentiis, M. Rinaldo, E. Esposito, G. Riccardi, A. Amore, M. Libra, G. Ciliberto, D.J.A. Jenkins, M. Montella.
  Journal of Cellular Physiology. September 30, 2016
  Hyperglycemia and hyperinsulinemia may play a role in breast carcinogenesis and prediabetes and diabetes have been associated with increased breast cancer (BC) risk. However, whether BC molecular subtypes may modify these associations is less clear. We therefore investigated these associations in all cases and by BC molecular subtypes among women living in Southern Italy. Cases were 557 patients with non‐metastatic incident BC and controls were 592 outpatients enrolled during the same period as cases and in the same hospital for skin‐related non‐malignant conditions. Adjusted multivariate logistic regression models were built to assess the risks of developing BC in the presence of prediabetes or diabetes. The analyses were repeated by strata of BC molecular subtypes: Luminal A, Luminal B, HER2+, and Triple Negative (TN). Prediabetes and diabetes were significantly associated with higher BC incidence after controlling for known risk factors (OR = 1.94, 95% CI 1.32–2.87 and OR = 2.46, 95% CI 1.38–4.37, respectively). Similar results were seen in Luminal A and B while in the TN subtype only prediabetes was associated with BC (OR = 2.43, 95% CI 1.11–5.32). Among HER2+ patients, only diabetes was significantly associated with BC risk (OR = 3.04, 95% CI 1.24–7.47). Furthermore, when postmenopausal HER2+ was split into hormone receptor positive versus negative, the association with diabetes remained significant only in the former (OR = 5.13, 95% CI 1.53–17.22). These results suggest that prediabetes and diabetes are strongly associated with BC incidence and that these metabolic conditions may be more relevant in the presence of breast cancer molecular subtypes with positive hormone receptors. J. Cell. Physiol. 232: 1144–1150, 2017. © 2016 Wiley Periodicals, Inc.

  September 30, 2016   doi: 10.1002/jcp.25579   open full text
• Hyperexcitability of Mesencephalic Trigeminal Neurons and Reorganization of Ion Channel Expression in a Rett Syndrome Model.
  Max F. Oginsky, Ningren Cui, Weiwei Zhong, Christopher M. Johnson, Chun Jiang.
  Journal of Cellular Physiology. September 30, 2016
  People with Rett syndrome (RTT) have defects in motor function also seen in Mecp2‐null mice. Motor function depends on not only central motor commands but also sensory feedback that is vulnerable to changes in excitability of propriosensory neurons. Here we report evidence for hyperexcitability of mesencephalic trigeminal (Me5) neurons in Mecp2‐null mice and a novel cellular mechanism for lowering its impact. In in vitro brain slices, the Me5 neurons in both Mecp2−/Y male and symptomatic Mecp2+/− female mice were overly excitable showing increased firing activity in comparison to their wild‐type (WT) male and asymptomatic counterparts. In Mecp2−/Y males, Me5 neurons showed a reduced firing threshold. Consistently, the steady‐state activation of voltage‐gated Na+ currents (INa) displayed a hyperpolarizing shift in the Mecp2‐null neurons with no change in the INa density. This seems to be due to NaV1.1, SCN1B and SCN4B overexpression and NaV1.2 and SCN3B under‐expression. In contrast to the hyperexcitability, the sag potential and postinhibitory rebound (PIR) were reduced in Mecp2‐null mice. In voltage‐clamp, the IH density was deficient by ∼33%, and the steady‐state half‐activation had a depolarizing shift of ∼10 mV in the Mecp2‐null mice. Quantitative PCR analysis indicated that HCN2 was decreased, HCN1 was upregulated with no change in HCN4 in Mecp2−/Y mice compared to WT. Lastly, blocking IH reduced the firing rate much more in WT than in Mecp2‐null neurons. These data suggest that the Mecp2 defect causes an increase in Me5 neuronal excitability likely attributable to alterations in INa, meanwhile IH is reduced likely altering neuronal excitability as well. J. Cell. Physiol. 232: 1151–1164, 2017. © 2016 Wiley Periodicals, Inc. Mesencephalic trigeminal V neurons are hyperexcitable in Mecp2‐null mice. Our data suggest that the Mecp2 defect that causes an increase in Me5 neuronal excitability is likely attributable to alterations in INa, meanwhile IH is reduced likely altering neuronal excitability as well. Further, the changes in these currents seem to be due to alterations in channel subunit composition.

  September 30, 2016   doi: 10.1002/jcp.25589   open full text
• Nephroprotective Effects of Metformin in Diabetic Nephropathy.
  Sreenithya Ravindran, Vinitha Kuruvilla, Kerry Wilbur, Shankar Munusamy.
  Journal of Cellular Physiology. September 26, 2016
  Metformin, a well‐known anti‐diabetic agent, is very effective in lowering blood glucose in patients with type 2 diabetes with minimal side‐effects. Metformin is also being recommended in the treatment of obesity and polycystic ovary syndrome. Metformin elicits its therapeutic effects mainly via activation of AMP‐activated kinase (AMPK) pathway. Renal cells under hyperglycemic or proteinuric conditions exhibit inactivation of cell defense mechanisms such as AMPK and autophagy, and activation of pathologic pathways such as mammalian target of rapamycin (mTOR), endoplasmic reticulum (ER) stress, epithelial‐to‐mesenchymal transition (EMT), oxidative stress, and hypoxia. As these pathologic pathways are intertwined with AMPK signaling, the potential benefits of metformin therapy in patients with type 2 diabetes would extend beyond its anti‐hyperglycemic effects. However, since metformin is eliminated unchanged through the kidneys and some studies have shown the incidence of lactic acidosis with its use during severe renal dysfunction, the use of metformin was contraindicated in patients with renal disease until recently. With more studies indicating the relatively low incidence of lactic acidosis and revealing the additional benefits with metformin therapy, the US FDA has now approved metformin to be administered in patients with established renal disease based on their renal function. The purpose of this review is to highlight the various mechanisms by which metformin protects renal cells that have lost its functionality in a diabetic or non‐diabetic setting and to enlighten the advantages and therapeutic potential of metformin as a nephroprotectant for patients with diabetic nephropathy and other non‐diabetic forms of chronic kidney disease. J. Cell. Physiol. 232: 731–742, 2017. © 2016 Wiley Periodicals, Inc. Metformin, a popular anti‐diabetic drug and an AMPK activator, modulates various signaling pathways altered in diabetic nephropathy and exhibits nephroprotective properties in various animal models of diabetes. Additional studies in humans are needed to ascertain its therapeutic potential as a nephroprotectant to treat nephropathy in patients with diabetes.

  September 26, 2016   doi: 10.1002/jcp.25598   open full text
• VIP Family Members Prevent Outer Blood Retinal Barrier Damage in a Model of Diabetic Macular Edema.
  Grazia Maugeri, Agata Grazia D'Amico, Caterina Gagliano, Salvatore Saccone, Concetta Federico, Sebastiano Cavallaro, Velia D'Agata.
  Journal of Cellular Physiology. September 21, 2016
  Diabetic macular edema (DME), characterized by an increase of thickness in the eye macular area, is due to breakdown of the blood‐retinal barrier (BRB). Hypoxia plays a key role in the progression of this pathology by activating the hypoxia‐inducible factors. In the last years, various studies have put their attention on the role of pituitary adenylate cyclase‐activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP) in retinal dysfunction. However, until now, no study has investigated their protective role against the harmful combined effect of both hyperglycemia and hypoxia on outer BRB. Therefore, in the present study, we have analyzed the role of these peptides on permeability, restoration of tight junctions expression and inhibition of hyperglycemia/hypoxia‐induced apoptosis, in an experimental in vitro model of outer BRB. Our results have demonstrated that the peptides' treatment have restored the integrity of outer BRB induced by cell exposure to hyperglycemia/hypoxia. Their effect is mediated through the activation of phosphoinositide 3 kinase (PI3K)/Akt and mammalian mitogen activated protein kinase/Erk kinase (MAPK/ERK) signaling pathways. In conclusion, our study further clarifies the mechanism through which PACAP and VIP perform the beneficial effect on retinal damage induced by hyperglycemic/hypoxic insult, responsible of DME progression. J. Cell. Physiol. 232: 1079–1085, 2017. © 2016 Wiley Periodicals, Inc. PACAP and VIP restore the integrity of outer BRB exposed to hyperglycemia/hypoxia. PACAP and VIP perform their beneficial effect on retinal damage through activation of PI3K/Akt and MAPK/ERK signaling cascades.

  September 21, 2016   doi: 10.1002/jcp.25510   open full text
• Effects of Conjugated Linoleic Acid Associated With Endurance Exercise on Muscle Fibres and Peroxisome Proliferator‐Activated Receptor γ Coactivator 1 α Isoforms.
  Rosario Barone, Claudia Sangiorgi, Antonella Marino Gammazza, Daniela D'Amico, Monica Salerno, Francesco Cappello, Cristoforo Pomara, Giovanni Zummo, Felicia Farina, Valentina Di Felice, Filippo Macaluso.
  Journal of Cellular Physiology. September 21, 2016
  Conjugated linoleic acid (CLA) has been reported to improve muscle hypertrophy, steroidogenesis, physical activity, and endurance capacity in mice, although the molecular mechanisms of its actions are not completely understood. The aim of the present study was to identify whether CLA alters the expression of any of the peroxisome proliferator‐activated receptor γ coactivator 1α (PGC1α) isoforms, and to evaluate the possible existence of fibre‐type‐specific hypertrophy in the gastrocnemius and plantaris muscles. Mice were randomly assigned to one of four groups: placebo sedentary, CLA sedentary, placebo trained, or CLA trained. The CLA groups were gavaged with 35 μl per day of Tonalin® FFA 80 food supplement containing CLA throughout the 6‐week experimental period, whereas the placebo groups were gavaged with 35 μl sunflower oil each day. Each administered dose of CLA corresponded to approximately 0.7 g/kg or 0.5%, of the dietary daily intake. Trained groups ran 5 days per week on a Rota‐Rod for 6 weeks at increasing speeds and durations. Mice were sacrificed by cervical dislocation and hind limb posterior muscle groups were dissected and used for histological and molecular analyses. Endurance training stimulated mitochondrial biogenesis by PGC1α isoforms (tot, α1, α2, and α3) but CLA supplementation did not stimulate PGC1α isoforms or mitochondrial biogenesis in trained or sedentary mice. In the plantaris muscle, CLA supplementation induced a fibre‐type‐specific hypertrophy of type IIx muscle fibres, which was associated with increased capillary density and was different from the fibre‐type‐specific hypertrophy induced by endurance exercise (of types I and IIb muscle fibres). J. Cell. Physiol. 232: 1086–1094, 2017. © 2016 Wiley Periodicals, Inc. CLA supplementation did not stimulate PGC1α isoforms or mitochondrial biogenesis in trained or sedentary mice. CLA supplementation induced a fibre‐type‐specific hypertrophy of type IIx muscle fibres.

  September 21, 2016   doi: 10.1002/jcp.25511   open full text
• Endothelial STAT3 Activation Increases Vascular Leakage Through Downregulating Tight Junction Proteins: Implications for Diabetic Retinopathy.
  Jang‐Hyuk Yun, Sung Wook Park, Kyung‐Jin Kim, Jong‐Sup Bae, Eun Hui Lee, Sun Ha Paek, Seung U. Kim, Sangkyu Ye, Jeong‐Hun Kim, Chung‐Hyun Cho.
  Journal of Cellular Physiology. September 21, 2016
  Vascular inflammation is characteristic feature of diabetic retinopathy. In diabetic retina, a variety of the pro‐inflammatory cytokines are elevated and involved in endothelial dysfunction. STAT3 transcription factor has been implicated in mediating cytokine signaling during vascular inflammation. However, whether and how STAT3 is involved in the direct regulation of the endothelial permeability is currently undefined. Our studies revealed that IL‐6‐induced STAT3 activation increases retinal endothelial permeability and vascular leakage in retinas of mice through the reduced expression of the tight junction proteins ZO‐1 and occludin. In a co‐culture model with microglia and endothelial cells under a high glucose condition, the microglia‐derived IL‐6 induced STAT3 activation in the retinal endothelial cells, leading to increasing endothelial permeability. In addition, IL‐6‐induced STAT3 activation was independent of ROS generation in the retinal endothelial cells. Moreover, we demonstrated that STAT3 activation downregulates the ZO‐1 and occludin levels and increases the endothelial permeability through the induction of VEGF production in retinal endothelial cells. These results suggest the potential importance of IL‐6/STAT3 signaling in regulating endothelial permeability and provide a therapeutic target to prevent the pathology of diabetic retinopathy. J. Cell. Physiol. 232: 1123–1134, 2017. © 2016 Wiley Periodicals, Inc. Vascular inflammation is characteristic feature of diabetic retinopathy. We demonstrated that STAT3 in retinal endothelial cells plays a causative role in downregulating ZO‐1 and occludin levels and increasing the endothelial permeability through VEGF production in retinal endothelial cells. These results suggest the potential importance of IL‐6/STAT3 signaling in regulating endothelial permeability and provide a therapeutic target to prevent the pathology of diabetic retinopathy.

  September 21, 2016   doi: 10.1002/jcp.25575   open full text
• HMGB1 Inhibits Apoptosis Following MI and Induces Autophagy via mTORC1 Inhibition.
  Eleonora Foglio, Giovanni Puddighinu, Antonia Germani, Matteo A. Russo, Federica Limana.
  Journal of Cellular Physiology. September 21, 2016
  Exogenous High Mobility Group Box‐1 protein (HMGB1) has been reported to protect the infarcted heart but the underlying mechanism is quite complex. In particular, its effect on ischemic cardiomyocytes has been poorly investigated. Aim of the present study was to verify whether and how autophagy and apoptosis were involved in HMGB1‐induced heart repair following myocardial infarction (MI). HMGB1 (200 ng) or denatured HMGB1 were injected in the peri‐infarcted region of mouse hearts following acute MI. Three days after treatment, an upregulation of autophagy was detected in infarcted HMGB1 treated hearts compared to controls. Specifically, HMGB1 induced autophagy by significantly upregulating the protein expression of LC3, Beclin‐1, and Atg7 in the border zone. To gain further insights into the molecular mechanism of HMGB1‐mediated autophagy, WB analysis were performed in cardiomyocytes isolated from 3 days infarcted hearts in the presence and in the absence of HMGB1 treatment. Results showed that upregulation of autophagy by HMGB1 treatment was potentially related to activation of AMP‐activated protein kinase (AMPK) and inhibition of the mammalian target of rapamycin complex 1 (mTORC1). Accordingly, in these hearts, phospho‐Akt signaling pathway was inhibited. The induction of autophagy was accompanied by reduced cardiomyocyte apoptotic rate and decreased expression levels of Bax/Bcl‐2 and active caspase‐3 in the border zone of 3 days infarcted mice following HMGB1 treatment. We report the first in vivo evidence that HMGB1 treatment in a murine model of acute MI might induce cardiomyocyte survival through attenuation of apoptosis and AMP‐activated protein kinase‐dependent autophagy. J. Cell. Physiol. 232: 1135–1143, 2017. © 2016 Wiley Periodicals, Inc. Exogenous High Mobility Group Box‐1 protein (HMGB1) has been reported to protect the infarcted heart but the underlying mechanism is quite complex; in particular, its effect on ischemic cardiomyocytes has been poorly investigated. HMGB1 (200 ng) or denatured HMGB1 were injected in the peri‐infarcted region of mouse hearts following acute MI. We report the first in vivo evidence that HMGB1 treatment in a murine model of acute MI might induce cardiomyocyte survival through attenuation of apoptosis and AMP‐activated protein kinase‐dependent autophagy.

  September 21, 2016   doi: 10.1002/jcp.25576   open full text
• Functional Effects of Cigarette Smoke‐Induced Changes in Airway Smooth Muscle Mitochondrial Morphology.
  Bharathi Aravamudan, Michael Thompson, Gary C. Sieck, Robert Vassallo, Christina M. Pabelick, Y. S. Prakash.
  Journal of Cellular Physiology. September 21, 2016
  Long‐term exposure to cigarette smoke (CS) triggers airway hyperreactivity and remodeling, effects that involve airway smooth muscle (ASM). We previously showed that CS destabilizes the networked morphology of mitochondria in human ASM by regulating the expression of mitochondrial fission and fusion proteins via multiple signaling mechanisms. Emerging data link regulation of mitochondrial morphology to cellular structure and function. We hypothesized that CS‐induced changes in ASM mitochondrial morphology detrimentally affect mitochondrial function, leading to CS effects on contractility and remodeling. Here, ASM cells were exposed to 1% cigarette smoke extract (CSE) for 48 h to alter mitochondrial fission/fusion, or by inhibiting the fission protein Drp1 or the fusion protein Mfn2. Mitochondrial function was assessed via changes in bioenergetics or altered rates of proliferation and apoptosis. Our results indicate that both exposure to CS and inhibition of mitochondrial fission/fusion proteins affect mitochondrial function (i.e., energy metabolism, proliferation, and apoptosis) in ASM cells. In vivo, the airways in mice chronically exposed to CS are thickened and fibrotic, and the expression of proteins involved in mitochondrial function is dramatically altered in the ASM of these mice. We conclude that CS‐induced changes in mitochondrial morphology (fission/fusion balance) correlate with mitochondrial function, and thus may control ASM proliferation, which plays a central role in airway health. J. Cell. Physiol. 232: 1053–1068, 2017. © 2016 Wiley Periodicals, Inc. Long‐term exposure to cigarette smoke (CS) triggers airway hyperreactivity and remodeling, effects that involve airway smooth muscle (ASM). Previous studies show that CS destabilizes the networked morphology of mitochondria in human ASM. We hypothesized that CS‐induced changes in ASM mitochondrial morphology detrimentally affect mitochondrial function, leading to CS effects on contractility and remodeling.

  September 21, 2016   doi: 10.1002/jcp.25508   open full text
• Store‐Operated Ca2+ Entry in Oocytes Modulate the Dynamics of IP3‐Dependent Ca2+ Release From Oscillatory to Tonic.
  Raphaël Courjaret, Maya Dib, Khaled Machaca.
  Journal of Cellular Physiology. September 07, 2016
  Ca2+ signaling is ubiquitous and mediates various cellular functions encoded in its spatial, temporal, and amplitude features. Here, we investigate the role of store‐operated Ca2+ entry (SOCE) in regulating the temporal dynamics of Ca2+ signals in Xenopus oocytes, which can be either oscillatory or tonic. Oscillatory Ca2+ release from intracellular stores is typically observed at physiological agonist concentration. When Ca2+ release leads to Ca2+ store depletion, this triggers the activation of SOCE that translates into a low‐amplitude tonic Ca2+ signal. SOCE has also been implicated in fueling Ca2+ oscillations when activated at low levels. Here, we show that sustained SOCE activation in the presence of IP3 to gate IP3 receptors (IP3R) results in a pump‐leak steady state across the endoplasmic reticulum (ER) membrane that inhibits Ca2+ oscillations and produces a tonic Ca2+ signal. Tonic signaling downstream of SOCE activation relies on focal Ca2+ entry through SOCE ER‐plasma membrane (PM) junctions, Ca2+ uptake into the ER, followed by release through open IP3Rs at distant sites, a process we refer to as “Ca2+ teleporting.” Therefore, sustained SOCE activation in the presence of an IP3‐dependent “leak” pathway at the ER membrane results in a switch from oscillatory to tonic Ca2+ signaling. J. Cell. Physiol. 232: 1095–1103, 2017. © 2016 Wiley Periodicals, Inc. Calcium influx through SOCE inhibits calcium oscillations and favors a tonic mode of signaling. This is achieved through focal calcium entry at SOCE clusters, followed by calcium uptake into the ER through SERCA and re‐release through open IP3 Receptors. A process referred to as “calcium teleporting.”

  September 07, 2016   doi: 10.1002/jcp.25513   open full text
• MiR‐145 Regulates Lipogenesis in Goat Mammary Cells Via Targeting INSIG1 and Epigenetic Regulation of Lipid‐Related Genes.
  Hui Wang, Huaiping Shi, Jun Luo, Yongqing Yi, Dawei Yao, Xueying Zhang, Gongzhen Ma, Juan J. Loor.
  Journal of Cellular Physiology. September 07, 2016
  MicroRNAs (miRNAs) are noncoding RNA molecules that regulate gene expression at the post‐transcriptional level to cause translational repression or degradation of targets. The profiles of miRNAs across stages of lactation in small ruminant species such as dairy goats is unknown. A small RNA library was constructed using tissue samples from mammary gland of Saanen dairy goats harvested at mid‐lactation followed by sequencing via Solexa technology. A total of 796 conserved miRNAs, 263 new miRNAs, and 821 pre‐miRNAs were uncovered. After comparative analyses of our sequence data with published mammary gland transcriptome data across different stages of lactation, a total of 37 miRNAs (including miR‐145) had significant differences in expression over the lactation cycle. Further studies revealed that miR‐145 regulates metabolism of fatty acids in goat mammary gland epithelial cells (GMEC). Compared with nonlactating mammary tissue, lactating mammary gland had a marked increase in expression of miR‐145. Overexpression of miR‐145 increased transcription of genes associated with milk fat synthesis resulting in greater fat droplet formation, triacylglycerol accumulation, and proportion of unsaturated fatty acids. In contrast, silencing of miR‐145 impaired fatty acid synthesis. Inhibition of miR‐145 increased methylation levels of fatty acid synthase (FASN), stearoyl‐CoA desaturase 1 (SCD1), peroxisome proliferator‐activated receptor gamma (PPARG), and sterol regulatory element binding transcription factor 1 (SREBF1). Luciferase reporter assays confirmed that insulin induced gene 1 (INSIG1) is a direct target of miR‐145. These findings underscore the need for further studies to evaluate the potential for targeting miR‐145 for improving beneficial milk components in ruminant milk. J. Cell. Physiol. 232: 1030–1040, 2017. © 2016 Wiley Periodicals, Inc. Mammary lipogenesis is partly regulated by epigenetic mechanisms through miR‐145 and INSIG1.

  September 07, 2016   doi: 10.1002/jcp.25499   open full text
• Hypoxia Modulates the Swelling‐Activated Cl Current in Human Glioblastoma Cells: Role in Volume Regulation and Cell Survival.
  Luigi Sforna, Marta Cenciarini, Silvia Belia, Antonio Michelucci, Mauro Pessia, Fabio Franciolini, Luigi Catacuzzeno.
  Journal of Cellular Physiology. August 26, 2016
  The malignancy of glioblastoma multiforme (GBM), the most common human brain tumor, correlates with the presence of hypoxic areas, but the underlying mechanisms are unclear. GBM cells express abundant Cl channels whose activity supports cell volume and membrane potential changes, ultimately leading to cell proliferation, migration, and escaping death. In non‐tumor tissues Cl channels are modulated by hypoxia, which prompted us to verify whether hypoxia would also modulate Cl channels in GBM cells. Our results show that in GBM cell lines, acute application of a hypoxic solution activates a Cl current displaying the biophysical and pharmacological features of the swelling‐activated Cl current (ICl,swell). We also found that acute hypoxia increased the cell volume by about 20%, and a 30% hypertonic solution partially inhibited the hypoxia‐activated Cl current, suggesting that cell swelling and the activation of the Cl current are sequential events. Notably, the hypoxia‐induced cell swelling was followed by a regulatory volume decrease (RVD) mediated mainly by ICl,swell. Since, a hypoxia‐induced prolonged cell swelling is usually regarded as a death insult, we hypothesized that the hypoxia‐activated Cl current could limit cell swelling and prevent necrotic death of GBM cells under hypoxic conditions. In accordance, we found that the ICl,swell inhibitor DCPIB hampered the RVD process, and more importantly it sensibly increased the hypoxia‐induced necrotic death in these cells. Taken together, these results suggest that Cl channels are strongly involved in the survival of GBM cells in a hypoxic environment, and may thus represent a new therapeutic target for this malignant tumor. J. Cell. Physiol. 232: 91–100, 2017. © 2016 Wiley Periodicals, Inc.

  August 26, 2016   doi: 10.1002/jcp.25393   open full text
• Extracellular Proteasomes Are Deficient in 19S Subunits as Revealed by iTRAQ Quantitative Proteomics.
  Anna S. Tsimokha, Julia J. Zaykova, Andrew Bottrill, Nikolai A. Barlev.
  Journal of Cellular Physiology. August 16, 2016
  Proteasome‐mediated proteolysis is critical for regulation of vast majority of cellular processes. In addition to their well‐documented functions in the nucleus and cytoplasm proteasomes have also been found in extracellular space. The origin and functions of these proteasomes, dubbed as circulating/plasmatic or extracellular proteasomes, are unclear. To gain insights into the molecular and functional differences between extracellular (EPs) and cellular proteasomes (CPs) we compared their subunit composition using iTRAQ‐based quantitative proteomics (iTRAQ LC/MS‐MS). Our analysis of purified from K562 cells or conditioned medium intact proteasome complexes led to an identification and quantification of 114 proteins, out of which 19 were 26S proteasome proteins (all subunits of the 20S proteasome and a small number of the 19S regulatory particle proteins), and 3 belonged to the ubiquitin system. Sixty‐two of proteasome interacting proteins (PIPs) were differentially represented in CP versus EP, with folds difference ranging from 1.5 to 4.8. The bioinformatics analysis revealed that functionally most of EP‐PIPs were associated with protein biosynthesis and, unlike CP‐PIPs, were under represented by chaperon/ATP‐binding proteins. Identities of some of the proteasome proteins and PIPs were verified by Western blotting. Importantly, we uncovered that the stoichiometry of the 20S versus 19S complexes in the extracellular proteasomes was different compared to the one calculated for the intracellular proteasomes. Specifically, the EP prep contained only three 19S subunits versus at least 18 in the CP one, suggesting that the extracellular proteasomes are deficient in 19S complexes, which may imply that they have special biological functions. J. Cell. Physiol. 232: 842–851, 2017. © 2016 Wiley Periodicals, Inc. To gain insights into the molecular and functional differences between extracellular (EPs) and cellular proteasomes (CPs) we compared their subunit composition using iTRAQ‐based quantitative proteomics (iTRAQ LC/MS‐MS). Importantly, we uncovered that the stoichiometry of the 20S versus 19S complexes in the extracellular proteasomes was different compared to the one calculated for the intracellular proteasomes, which implies that the two species of proteasomes may have different biological functions.

  August 16, 2016   doi: 10.1002/jcp.25492   open full text
• Uric Acid Amplifies Aβ Amyloid Effects Involved in the Cognitive Dysfunction/Dementia: Evidences From an Experimental Model In Vitro.
  Giovambattista Desideri, Roberta Gentile, Andrea Antonosante, Elisabetta Benedetti, Davide Grassi, Loredana Cristiano, Antonello Manocchio, Sara Selli, Rodolfo Ippoliti, Claudio Ferri, Claudio Borghi, Antonio Giordano, Annamaria Cimini.
  Journal of Cellular Physiology. August 16, 2016
  There is still a considerable debate concerning whether uric acid is neuroprotective or neurotoxic agent. To clarify this topic, we tested the effects of uric acid on neuronal cells biology by using differentiated SHSY5Y neuroblastoma cells incubated with amyloid β to reproduce an in vitro model of Alzheimer's disease. The incubation of cells with uric acid at the dose of 40 µM or higher significantly reduced cell viability and potentiated the proapoptotic effect of amyloid β. Finally, uric acid enhanced the generation of 4‐hydroxynonenal and the expression of PPARβ/δ promoted by amyloid β, indicating a prooxidant effects. In conclusion, uric acid could exert a detrimental influence on neuronal biology being this influence further potentiated by the concomitant exposure to neurotoxic stimuli. This effect is evident for uric acid concentrations close to those achievable in cerebrospinal fluid in presence of mild hyperuricemia thus suggesting a potential role of uric acid in pathophysiology of cognitive dysfunction. These effects are influenced by the concentrations of uric acid and by the presence of favoring conditions that commonly occur in neurodegenerative disorders and well as in the aging brain, including increased oxidative stress and exposure to amyloid β. J. Cell. Physiol. 232: 1069–1078, 2017. © 2016 Wiley Periodicals, Inc. The study suggests that uric acid could exert detrimental effects on brain structure and function by directly influencing the viability of neuronal cells and their ability to establish synaptic connections.

  August 16, 2016   doi: 10.1002/jcp.25509   open full text
• Chondrocyte Morphology in Stiff and Soft Agarose Gels and the Influence of Fetal Calf Serum.
  Asima Karim, Andrew C. Hall.
  Journal of Cellular Physiology. August 16, 2016
  Changes to chondrocyte volume/morphology may have deleterious effects on extracellular matrix (ECM) metabolism potentially leading to cartilage deterioration and osteoarthritis (OA). The factors controlling chondrocyte properties are poorly understood, however, pericellular matrix (PCM) weakening may be involved. We have studied the density, volume, morphology, and clustering of cultured bovine articular chondrocytes within stiff (2% w/v) and soft (0.2% w/v) three‐dimensional agarose gels. Gels with encapsulated chondrocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM; fetal calf serum (FCS) 1–10%;380 mOsm) for up to 7 days. Chondrocytes were fluorescently labeled after 1, 3, and 7 days with 5‐chloromethylfluorescein‐diacetate (CMFDA) and propidium iodide (PI) or 1,5‐bis{[2‐(di‐methylamino)ethyl]amino}‐4,8‐dihydroxyanthracene‐9,10‐dione (DRAQ5) to identify cytoplasmic space or DNA and imaged by confocal laser scanning microscopy (CLSM). Chondrocyte density, volume, morphology, and clustering were quantified using Volocity™ software. In stiff gels after 7 d with 10% FCS, chondrocyte density remained unaffected and morphology was relatively normal with occasional cytoplasmic processes. However, in soft gels by day 1, chondrocyte volume increased (P = 0.0058) and by day 7, density increased (P = 0.0080), along with the percentage of chondrocytes of abnormal morphology (P < 0.0001) and enhanced clustering (P < 0.05), compared to stiff gels. FCS exacerbated changes to density (P < 0.01), abnormal morphology (P < 0.001) and clustering (P < 0.01) compared to lower concentrations at the same gel strength. Reduced gel stiffness and/or increased FCS concentrations promoted chondrocyte proliferation and clustering, increased cell volume, and stimulated abnormal morphology, producing similar changes to those occurring in OA. The increased penetration of factors in FCS into soft gels may be important in the development of these abnormal chondrocyte properties. J. Cell. Physiol. 232: 1041–1052, 2017. © 2016 Wiley Periodicals, Inc. Changes to chondrocyte properties occur during the process of osteoarthritis but their role in this disorder is poorly understood. We described a three‐dimensional culture model using stiff and soft agarose and quantified chondrocyte properties of density, morphology, volume, and clustering. Some of the changes we observed are very similar to those that occur in OA and the results suggested that the penetration of serum factors in the culture medium could be responsible.

  August 16, 2016   doi: 10.1002/jcp.25507   open full text
• Ascorbic Acid Induces Necrosis in Human Laryngeal Squamous Cell Carcinoma via ROS, PKC, and Calcium Signaling.
  Min‐Woo Baek, Heui‐Seung Cho, Sun‐Hun Kim, Won‐Jae Kim, Ji‐Yeon Jung.
  Journal of Cellular Physiology. August 16, 2016
  Ascorbic acid induces apoptosis, autophagy, and necrotic cell death in cancer cells. We investigated the mechanisms by which ascorbic acid induces death in laryngeal squamous cell carcinoma Hep2 cells. Ascorbic acid markedly reduced cell viability and induced death without caspase activation and an increase in cytochrome c. Hep2 cells exposed to ascorbic acid exhibited membrane rupture and swelling, the morphological characteristics of necrotic cell death. The generation of reactive oxygen species (ROS) was increased in Hep2 cells treated with ascorbic acid, and pretreatment with N‐acetylcysteine blocked ascorbic acid‐induced cell death. Ascorbic acid also stimulated protein kinase C (PKC) signaling, especially PKC α/β activation, and subsequently increased cytosolic calcium levels. However, ascorbic acid‐induced necrotic cell death was inhibited by Ro‐31‐8425 (PKC inhibitor) and BAPTA‐AM (cytosolic calcium‐selective chelator). ROS scavenger NAC inhibited PKC activation induced by ascorbic acid and Ro‐31‐8425 suppressed the level of cytosolic calcium increased by ascorbic acid, indicating that ROS is represented as an upstream signal of PKC pathway and PKC activation leads to the release of calcium into the cytosol, which ultimately regulates the induction of necrosis in ascorbic acid‐treated Hep2 cells. These data demonstrate that ascorbic acid induces necrotic cell death through ROS generation, PKC activation, and cytosolic calcium signaling in Hep2 cells. J. Cell. Physiol. 232: 417–425, 2017. © 2016 Wiley Periodicals, Inc. Ascorbic acid induced necrotic cell death in human laryngeal squamous cell carcinoma through ROS, PKC, and calcium signaling pathway.

  August 16, 2016   doi: 10.1002/jcp.25438   open full text
• 3‐Methylcholanthrene/Aryl‐Hydrocarbon Receptor‐Mediated Hypertension Through eNOS Inactivation.
  Chih‐Cheng Chang, Yung‐Ho Hsu, Hsiu‐Chu Chou, Yuan‐Chii G. Lee, Shu‐Hui Juan.
  Journal of Cellular Physiology. August 10, 2016
  Endothelial nitric oxide synthase (eNOS) modulates vascular blood pressure and is predominantly expressed in endothelial cells and activated through the protein kinase B (Akt/PKB)‐dependent pathway. We previously reported that 3‐methylcholanthrene (3MC) activates the aryl hydrocarbon receptor (AhR) and reduces PI3K/Akt phosphorylation. This study investigated the mechanism underlying the downregulatory effects of 3‐MC on nitric oxide (NO) production occurring through the AhR/RhoA/Akt‐mediated mechanism. The mechanism underlying the effects of 3‐MC on eNOS activity and blood pressure was examined in vitro and in vivo through genetic and pharmacological approaches. Results indicated that 3‐MC modified heat shock protein 90 (HSP90), caveolin‐1, dynein, and eNOS mRNA and protein expression through the AhR/RhoA‐dependent mechanism in mouse cerebral vascular endothelial cells (MCVECs) and that 3‐MC reduced eNOS phosphorylation through the AhR/RhoA‐mediated inactivation of Akt1. The upregulation of dynein expression was associated with decreased eNOS dimer formation (eNOS dimer; an activated form of the enzyme). Coimmunoprecipitation assay results indicated that 3‐MC significantly reduced the interaction between eNOS and its regulatory proteins, including Akt1 and HSP90, but increased the interaction between eNOS and caveolin‐1. Immunofluorescence and Western blot analysis revealed that 3‐MC reduced the amount of membrane‐bound activated eNOS, and a modified Griess assay revealed that 3‐MC concomitantly reduced NO production. However, simvastatin reduced 3‐MC‐mediated murine hypertension. Our study results indicate that AhR, RhoA, and eNOS have major roles in blood pressure regulation. Statin intervention might provide a potential therapeutic approach for reducing hypertension caused by 3‐MC. J. Cell. Physiol. 232: 1020–1029, 2017. © 2016 Wiley Periodicals, Inc. Our study is the first to report that the AhR ligand 3‐MC inactivated eNOS associated with downregulated HSP90 and upregulated caveolin‐1 and dynein expression through AhR/RhoA‐mediated PI3K/Akt inactivation, leading to hypertension in mice. The blockade of AhR and RhoA by using siAhR, DNRhoA, or statins reversed the inhibitory effects of 3‐MC on eNOS by correcting the alterations of eNOS regulatory proteins.

  August 10, 2016   doi: 10.1002/jcp.25497   open full text
• Differentiation of Keratinocytes Modulates Skin HPA Analog.
  Justyna M. Wierzbicka, Michał A. Żmijewski, Jakub Antoniewicz, Michal Sobjanek, Andrzej T. Slominski.
  Journal of Cellular Physiology. August 10, 2016
  It is well established, that epidermal keratinocytes express functional equivalent of hypothalamus–pituitary–adrenal axis (HPA) in order to respond to changing environment and maintain internal homeostasis. We are presenting data indicating that differentiation of primary neonatal human keratinocytes (HPEKp), induced by prolonged incubation or calcium is accompanied by significant changes in the expression of the elements of skin analog of HPA (sHPA). Expression of CRF, UCN1‐3, POMC, ACTH, CRFR1, CRFR2, MC1R, MC2R, and GR (coded by NR3C1 gene) were observed on gene/protein levels along differentiation of keratinocytes in culture with similar pattern seen by immunohistochemistry on full thickness skin biopsies. Expression of CRF was more pronounced in less differentiated keratinocytes, which corresponded to the detection of CRF immunoreactivity preferentially in the stratum basale. POMC expression was enhanced in more differentiated keratinocytes, which corresponded to detection of ACTH immunoreactivity, predominantly in the stratum spinosum and stratum granulosum. Expression of urocortins was also affected by induction of HPEKp differentiation. Immunohistochemical studies showed high prevalence of CRFR1 in well differentiated keratinocytes, while smaller keratinocytes showed predominantly CRFR2 immunoreactivity. MC2R mRNA levels were elevated from days 4 to 8 of in vitro incubation, while MC2R immunoreactivity was the highest in the upper layers of epidermis. Similar changes in mRNA/protein levels of sHPA elements were observed in HPEKp keratinocytes treated with calcium. Summarizing, preferential expression of CRF and POMC (ACTH) by populations of keratinocytes on different stage of differentiation resembles organization of central HPA axis suggesting their distinct role in physiology and pathology of the epidermis. J. Cell. Physiol. 232: 154–166, 2017. © 2016 Wiley Periodicals, Inc. It is well established, that epidermal keratinocytes express functional equivalent of hypothalamus–pituitary–adrenal axis (HPA) in order to respond to changing environment and maintain internal homeostasis. Here, we are showing for the first time preferential expression of HPA axis elements by populations of keratinocytes on different stage of differentiation that resembles organization of central HPA axis suggesting their distinct role in physiology and pathology of the epidermis.

  August 10, 2016   doi: 10.1002/jcp.25400   open full text
• Tissue Inhibitor of Metalloproteinase 1 Influences Vascular Adaptations to Chronic Alterations in Blood Flow.
  Erin R. Mandel, Cassandra Uchida, Emmanuel Nwadozi, Armin Makki, Tara L. Haas.
  Journal of Cellular Physiology. July 28, 2016
  Remodeling of the skeletal muscle microvasculature involves the coordinated actions of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitor of metalloproteinases (TIMPs). We hypothesized that the loss of TIMP1 would enhance both ischemia and flow‐induced vascular remodeling by increasing MMP activity. TIMP1 deficient (Timp1−/−) and wild‐type (WT) C57BL/6 mice underwent unilateral femoral artery (FA) ligation or were treated with prazosin, an alpha‐1 adrenergic receptor antagonist, in order to investigate vascular remodeling to altered flow. Under basal conditions, Timp1−/− mice had reduced microvascular content as compared to WT mice. Furthermore, vascular remodeling was impaired in Timp1−/− mice. Timp1−/− mice displayed reduced blood flow recovery in response to FA ligation and no arteriogenic response to prazosin treatment. Timp1−/− mice failed to undergo angiogenesis in response to ischemia or prazosin, despite maintaining the capacity to increase VEGF‐A and eNOS mRNA. Vascular permeability was increased in muscles of Timp1−/− mice in response to both prazosin treatment and FA ligation, but this was not accompanied by greater MMP activity. This study highlights a previously undescribed integral role for TIMP1 in both vascular network maturation and adaptations to ischemia or alterations in flow. J. Cell. Physiol. 232: 831–841, 2017. © 2016 Wiley Periodicals, Inc. Remodeling of the skeletal muscle microvasculature involves the coordinated actions of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitor of metalloproteinases (TIMPs). This study highlights a previously undescribed integral role for TIMP1 in vascular network maturation and in adaptations to altered blood flow.

  July 28, 2016   doi: 10.1002/jcp.25491   open full text
• Coumestrol Inhibits Proliferation and Migration of Prostate Cancer Cells by Regulating AKT, ERK1/2, and JNK MAPK Cell Signaling Cascades.
  Whasun Lim, Muhah Jeong, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. July 28, 2016
  Coumestrol is the one of the major phytoestrogens which is abundant in soybeans, legumes, brussel sprouts, and spinach. The beneficial effects of coumestrol are well known in various biological processes including; neuroprotective effects on the nervous system, function of the female reproductive system, anti‐bacterial properties, and anti‐cancer effects. Although the anti‐tumor activity of coumestrol has been demonstrated for ovarian, breast, lung, and cervical cancers, little is known of its effects on prostate cancer. Therefore, in the present study, we investigated the chemotherapeutic effects of coumestrol on two prostate cancer cell lines, PC3 and LNCaP. Our results showed that coumestrol decreased proliferation and migration and induced apoptosis in both PC3 and LNCaP cells. Moreover, effects of coumestrol on cell signaling pathways were investigated and it increased phosphorylation of ERK1/2, JNK, P90RSK, and P53 proteins in a dose‐ and time‐dependent manner whereas phosphorylation of AKT was reduced by coumestrol under the same conditions for culture of PC3 and LNCaP cells. In addition, mitochondrial dysfunction was induced by coumestrol as evidenced by a significant loss of mitochondrial membrane potential. Furthermore, cleavage of caspase‐3 and caspase‐9, the apoptotic proteins associated with mitochondria, also changed in response to coumestrol. Coumestrol also caused mitochondrial dysfunction resulting in an increase in ROS production in PC3 and LNCaP cells. These results suggest that coumestrol can inhibit progression of prostate cancer and may be a novel chemotherapeutic agent for treatment of prostate cancer via effects mediated via the PI3K/AKT and ERK1/2 and JNK MAPK cell signaling pathways. J. Cell. Physiol. 232: 862–871, 2017. © 2016 Wiley Periodicals, Inc. Our results indicated that coumestrol can inhibit progression of prostate cancer cells by affecting their viability and mitochondrial functions, and may be a novel chemotherapeutic agent for treatment of prostate cancer via effects mediated via the PI3K/AKT and ERK1/2 and JNK MAPK cell signal transduction cascades.

  July 28, 2016   doi: 10.1002/jcp.25494   open full text
• Nestin Expressed by Pre‐Existing Cardiomyocytes Recapitulated in Part an Embryonic Phenotype; Suppressive Role of p38 MAPK.
  Marc‐Andre Meus, Vanessa Hertig, Louis Villeneuve, Jean‐Francois Jasmin, Angelino Calderone.
  Journal of Cellular Physiology. July 27, 2016
  Nestin(+)‐cardiomyocytes were identified in the ischemically damaged human/rodent heart, albeit the cellular source, and signaling events implicated in the appearance of the intermediate filament protein remained undefined. Expression of the enhanced green fluorescent protein (EGFP) driven by the second intron of the nestin gene identified a subpopulation of EGFP/nestin(+) cells that differentiated to a vascular phenotype in the peri‐infarct/infarct region of post‐MI mice albeit the transgene was not detected in nestin(+)‐cardiomyocytes. α‐MHC‐driven expression of the reporter mCherry was detected in troponin‐T(+)‐ and nestin(+)‐cardiomyocytes in the peri‐infarct/infarct region of post‐MI mice. However, the cell cycle re‐entry of nestin/mCherry(+)‐cardiomyocytes was not observed. Nestin staining was identified in a paucity of neonatal rat ventricular cardiomyocytes (NNVM). Exposure to phorbol 12,13‐dibutyrate (PDBu) induced NNVM hypertrophy but did not promote nestin expression or Brdu incorporation. PDBu treatment of NNVMs phosphorylated p38 MAPK and HSP27 and HSP27 phosphorylation was abrogated by the p38 MAPK inhibitor SB203580. PDBu/SB203580 co‐treatment significantly increased the percentage of NNVMs that expressed nestin and incorporated Brdu. In the heart of embryonic 10.5 day mice, nestin immunoreactivity was observed in cycling troponin‐T(+)‐cardiomyocytes. Nestin was also detected in embryonic rat ventricular cardiomyocytes and depletion of the intermediate filament protein attenuated cell cycle re‐entry. Thus, nestin expressed by pre‐existing cardiomyocytes following ischemic damage recapitulated in part an embryonic trait and may provide the requisite phenotype to initiate cell cycle re‐entry. However, the overt activation of the p38 MAPK pathway post‐MI may in part limit the appearance and inhibit the cell cycle re‐entry of nestin(+)‐cardiomyocytes. J. Cell. Physiol. 232: 1717–1727, 2017. © 2016 Wiley Periodicals, Inc. Nestin was expressed by pre‐existing cardiomyocytes following ischemic damage to the adult heart and recapitulated in part an embryonic phenotype. However, nestin‐expressing cardiomyocytes were unable to re‐enter the cell cycle following ischemic damage because of the suppressive effect of the p38 MAPK pathway.

  July 27, 2016   doi: 10.1002/jcp.25496   open full text
• Late Endosomal Recycling of Open MHC‐I Conformers.
  Hana Mahmutefendić, Gordana Blagojević Zagorac, Kristina Grabušić, Ljerka Karleuša, Senka Maćešić, Frank Momburg, Pero Lučin.
  Journal of Cellular Physiology. July 27, 2016
  With an increasing number of endosomal cargo molecules studied, it is becoming clear that endocytic routes are diverse, and the cell uses more pathways to adjust expression of cell surface proteins. Intracellular itinerary of integral membrane proteins that avoid the early endosomal recycling route is not enough studied. Therefore, we studied endocytic trafficking of empty Ld (eLd) molecules, an open form of murine MHC‐I allele, in fibroblast‐like cells. Pulse labeling of cell surface eLd with mAbs and internalization kinetics suggest two steps of endosomal recycling: rapid and late. The same kinetics was also observed for human open MHC‐I conformers. Kinetic modeling, using in‐house developed software for multicompartment analysis, colocalization studies and established protocols for enriched labeling of the late endosomal (LE) pool of eLd demonstrated that the late step of recycling occurs from an LE compartment. Although the majority of eLd distributed into pre‐degradative multivesicular bodies (MVBs), these LE subsets were not a source for eLd recycling. The LE recycling of eLd did not require Rab7 membrane domains, as demonstrated by Rab7‐silencing, but required vectorial LE motility, suggesting that LE recycling occurs from dynamic tubulovesicular LE domains prior segregation of eLd in MVBs. Thus, our study indicates that LE system should not be simply considered as a feeder for loading of the degradative tract of the cell but also as a feeder for loading of the plasma membrane and thereby contribute to the maintenance of homeostasis of plasma membrane proteins. J. Cell. Physiol. 232: 872–887, 2017. © 2016 Wiley Periodicals, Inc. Study of endosomal trafficking of open MHC‐I conformers reveals that endosomal recycling may occur from late endosomes, suggesting that the late endosomal system should not be simply considered as a feeder for loading of the degradative tract of the cell.

  July 27, 2016   doi: 10.1002/jcp.25495   open full text
• Tyrosine Residues Regulate Multiple Nuclear Functions of P54nrb.
  Ahn R. Lee, Wayne Hung, Ning Xie, Liangliang Liu, Leye He, Xuesen Dong.
  Journal of Cellular Physiology. July 27, 2016
  The non‐POU‐domain‐containing octamer binding protein (NONO; also known as p54nrb) has various nuclear functions ranging from transcription, RNA splicing, DNA synthesis and repair. Although tyrosine phosphorylation has been proposed to account for the multi‐functional properties of p54nrb, direct evidence on p54nrb as a phosphotyrosine protein remains unclear. To investigate the tyrosine phosphorylation status of p54nrb, we performed site‐directed mutagenesis on the five tyrosine residues of p54nrb, replacing the tyrosine residues with phenylalanine or alanine, and immunoblotted for tyrosine phosphorylation. We then preceded with luciferase reporter assays, RNA splicing minigene assays, co‐immunoprecipitation, and confocal microscopy to study the function of p54nrb tyrosine residues on transcription, RNA splicing, protein–protein interaction, and cellular localization. We found that p54nrb was not phosphorylated at tyrosine residues. Rather, it has non‐specific binding affinity to anti‐phosphotyrosine antibodies. However, replacement of tyrosine with phenylalanine altered p54nrb activities in transcription co‐repression and RNA splicing in gene context‐dependent fashions by means of differential regulation of p54nrb protein association with its interacting partners and co‐regulators of transcription and splicing. These results demonstrate that tyrosine residues, regardless of phosphorylation status, are important for p54nrb function. J. Cell. Physiol. 232: 852–861, 2017. © 2016 Wiley Periodicals, Inc. P54nrb (aka NONO) has various nuclear functions ranging from transcription, RNA splicing, DNA synthesis and repair. Although Tyr phosphorylation has been proposed to account for the multi‐functional properties of p54nrb, direct evidence on p54nrb as a phosphotyrosine protein remains unclear. Our studies demonstrate that the Tyr residues of p54nrb are not phosphorylated, but nevertheless significantly regulate p54nrb function by means of potential temporal and spatial regulation of p54nrb, resulting in altered interaction of p54nrb with various co‐regulatory splicing or transcriptional proteins.

  July 27, 2016   doi: 10.1002/jcp.25493   open full text
• Atrogin‐1 Increases Smooth Muscle Contractility Through Myocardin Degradation.
  Pavneet Singh, Dong Li, Yu Gui, Xi‐Long Zheng.
  Journal of Cellular Physiology. July 21, 2016
  Atrogin‐1, an E3 ligase present in skeletal, cardiac and smooth muscle, down‐regulates myocardin protein during skeletal muscle differentiation. Myocardin, the master regulator of smooth muscle cell (SMC) differentiation, induces expression of smooth muscle marker genes through its association with serum response factor (SRF), which binds to the CArG box in the promoter. Myocardin undergoes ubiquitylation and proteasomal degradation. Evidence suggests that proteasomal degradation of myocardin is critical for myocardin to exert its transcriptional activity, but there is no report about the E3 ligase responsible for myocardin ubiquitylation and subsequent transactivation. Here, we showed that overexpression of atrogin‐1 increased contractility of cultured SMCs and mouse aortic tissues in organ culture. Overexpression of dominant‐negative myocardin attenuated the increase in SMC contractility induced by atrogin‐1. Atrogin‐1 overexpression increased expression of the SM contractile markers while downregulated expression of myocardin protein but not mRNA. Atrogin‐1 also ubiquitylated myocardin for proteasomal degradation in vascular SMCs. Deletion studies showed that atrogin‐1 directly interacted with myocardin through its amino acids 284–345. Immunostaining studies showed nuclear localization of atrogin‐1, myocardin, and the Rpt6 subunit of the 26S proteasome. Atrogin‐1 overexpression not only resulted in degradation of myocardin but also increased recruitment of RNA Polymerase II onto the promoters of myocardin target genes. In summary, our results have revealed the roles for atrogin‐1 in the regulation of smooth muscle contractility through enhancement of myocardin ubiquitylation/degradation and its transcriptional activity. J. Cell. Physiol. 232: 806–817, 2017. © 2016 Wiley Periodicals, Inc. Atrogin‐1 is an E3 ligase present in skeletal, cardiac and smooth muscle. Our study has shown that atrogin‐1 down‐regulates myocardin protein in human vascular smooth muscle cells, but increases transcriptional activity of myocardin on its target genes.

  July 21, 2016   doi: 10.1002/jcp.25485   open full text
• Integrated Transcriptome Map Highlights Structural and Functional Aspects of the Normal Human Heart.
  Maria Caracausi, Allison Piovesan, Lorenza Vitale, Maria Chiara Pelleri.
  Journal of Cellular Physiology. July 21, 2016
  A systematic meta‐analysis of the available gene expression profiling datasets for the whole normal human heart generated a quantitative transcriptome reference map of this organ. Transcriptome Mapper (TRAM) software integrated 32 gene expression profile datasets from different sources returning a reference value of expression for each of the 43,360 known, mapped transcripts assayed by any of the experimental platforms used in this regard. Main findings include the visualization at the gene and chromosomal levels of the classical description of the basic histology and physiology of the heart, the identification of suitable housekeeping reference genes, the analysis of stoichiometry of gene products, and the focusing on chromosome 21 genes, which are present in one excess copy in Down syndrome subjects, presenting cardiovascular defects in 30–40% of cases. Independent in vitro validation showed an excellent correlation coefficient (r = 0.98) with the in silico data. Remarkably, heart/non‐cardiac tissue expression ratio may also be used to anticipate that effects of mutations will most probably affect or not the heart. The quantitative reference global portrait of gene expression in the whole normal human heart illustrates the structural and functional aspects of the whole organ and is a general model to understand the mechanisms underlying heart pathophysiology. J. Cell. Physiol. 232: 759–770, 2017. © 2016 Wiley Periodicals, Inc. A systematic meta‐analysis of the available gene expression profiling datasets for the whole normal human heart retrieved 32 microarray experiments performed on the whole organ. These data were integrated through the Transcriptome Mapper (TRAM) software to obtain a reference typical value of expression for each of the 43,360 known, mapped transcripts. In vitro validation through real‐time RT‐PCR showed an excellent correlation coefficient (r = 0.98) with the in silico data.

  July 21, 2016   doi: 10.1002/jcp.25471   open full text
• Multifaceted Breast Cancer: The Molecular Connection With Obesity.
  Antonia Feola, Serena Ricci, Soumaya Kouidhi, Antonietta Rizzo, Antonella Penon, Pietro Formisano, Antonio Giordano, Angelina Di Carlo, Marina Di Domenico.
  Journal of Cellular Physiology. July 21, 2016
  Obesity is characterized by a disruption in energy balance regulation that results in an excess accumulation of body fat. Its increasing prevalence poses a major public health concern because it is a risk factor for a host of additional chronic conditions, including type 2 diabetes, hypertension, and cardiovascular disease. Obesity is increasingly recognized as a growing cause of cancer risk. In particular excessive adipose expansion during obesity causes adipose dysfunction and inflammation that can regulate tumor growth. In obesity, dysregulated systemic metabolism and inflammation induce hyperinsulinemia, hyperglycemia, dyslipidemia, and enhance sex hormone production with increased secretion of proinflammatory adipokine that impact breast cancer development and progression. This review describes how adipose inflammation that characterizes obesity is responsible of microenvironment to promote cancer, and discuss how steroid hormones, that are essential for the maintenance of the normal development, growth and differentiation of the cells, influence the induction and progression of breast cancer. J. Cell. Physiol. 232: 69–77, 2017. © 2016 Wiley Periodicals, Inc. Obesity is increasingly recognized as a growing cause of cancer risk. Excessive adipose expansion during obesity causes adipose dysfunction and inflammation that can regulate tumor growth. In obesity, dysregulated systemic metabolism and inflammation induce hyperinsulinemia.

  July 21, 2016   doi: 10.1002/jcp.25475   open full text
• Expression of P2 Purinergic Receptors in Mesenchymal Stem Cells and Their Roles in Extracellular Nucleotide Regulation of Cell Functions.
  Lin‐Hua Jiang, Yunjie Hao, Fatema Mousawi, Hongsen Peng, Xuebin Yang.
  Journal of Cellular Physiology. July 20, 2016
  Extracellular ATP and other nucleotides induce autocrine and/or paracrine purinergic signalling via activation of the P2 receptors on the cell surface, which represents one of the most common signalling mechanisms. Mesenchymal stem cells (MSC) are a type of multipotent adult stem cells that have many promising applications in regenerative medicine. There is increasing evidence to show that extracellular nucleotides regulate MSC functions and P2 receptor‐mediated purinergic signalling plays an important role in such functional regulation. P2 receptors comprise ligand‐gated ion channel P2X receptors and G‐protein‐coupled P2Y receptors. In this review, we provide an overview of the current understanding with respect to expression of the P2X and P2Y receptors in MSC and their roles in mediating extracellular nucleotide regulation of MSC proliferation, migration and differentiation. J. Cell. Physiol. 232: 287–297, 2017. © 2016 Wiley Periodicals, Inc.

  July 20, 2016   doi: 10.1002/jcp.25484   open full text
• Vimentin as a Marker of Early Differentiating, Highly Motile Corneal Epithelial Cells.
  Federico Castro‐Muñozledo, Diana G. Meza‐Aguilar, Rocío Domínguez‐Castillo, Veremundo Hernández‐Zequinely, Erika Sánchez‐Guzmán.
  Journal of Cellular Physiology. July 20, 2016
  Vimentin (Vim), a cytoskeletal intermediate filament, is part of a naturally occurring reversible program, the Epithelial‐Mesenchymal Transition (EMT), which converts epithelial cells into mesenchymal‐like derivatives. Based on previous results showing that epithelial cells co‐express Vim and keratin (Krt) as part of a cytoskeletal network which confers them a highly motile phenotype, we explored the role of Vim in rabbit corneal epithelial cells or RCE1(5T5) cells, an established model of corneal epithelial differentiation. Vim and keratin filaments were co‐expressed in cells localized at the proliferative/migratory rim of the growing colonies, but not in basal cells from the center of the colonies nor at suprabasal cell layers. Flow cytometry and qPCR demonstrated that there was a decrease in Krt+/Vim+ cell number and ΔNp63α expression when cells reached confluence and formed a 4–5 layered epithelium, while there was a concomitant increase of both Pax‐6 expression and Krt+/Vim− cells. Inhibition of cell proliferation with mitomycin C did not modify cell motility nor the expression of Vim. We studied the distribution and expression of α6 integrin, a protein also involved in cell migration. The results demonstrated that α6 integrin had a distribution which was, in part, co‐linear with Vim at the proliferative/migratory rim of cell colonies, suggesting an indirect interaction between these proteins. Immunoprecipitation and immunostaining assays indicated that plectin might be mediating such interaction. These data suggest that Vim expression in corneal epithelium is found in a cell population composed of highly motile cells with a Vim+/Krt+/ΔNp63α+/Pax‐6low/α6 integrin+ phenotype. J. Cell. Physiol. 232: 818–830, 2017. © 2016 Wiley Periodicals, Inc. During growth and wound healing, highly motile epithelial cells require the expression of a specific cytoskeletal network to sustain organelle translocation during cell movement. Such requirements might be fulfilled by the copolymerization of Vim and Krt at the migratory edges of cells, and its interaction with Extracellular Matrix, a task achieved by relocation of alpha6beta4 integrin into focal adhesion plaques, and its association with plectin as an integrator molecule.

  July 20, 2016   doi: 10.1002/jcp.25487   open full text
• Evidence for the Role of BAG3 in Mitochondrial Quality Control in Cardiomyocytes.
  Farzaneh G. Tahrir, Tijana Knezevic, Manish K. Gupta, Jennifer Gordon, Joseph Y. Cheung, Arthur M. Feldman, Kamel Khalili.
  Journal of Cellular Physiology. July 19, 2016
  Mitochondrial abnormalities impact the development of myofibrillar myopathies. Therefore, understanding the mechanisms underlying the removal of dysfunctional mitochondria from cells is of great importance toward understanding the molecular events involved in the genesis of cardiomyopathy. Earlier studies have ascribed a role for BAG3 in the development of cardiomyopathy in experimental animals leading to the identification of BAG3 mutations in patients with heart failure which may play a part in the onset of disease development and progression. BAG3 is co‐chaperone of heat shock protein 70 (HSP70), which has been shown to modulate apoptosis and autophagy, in several cell models. In this study, we explore the potential role of BAG3 in mitochondrial quality control. We demonstrate that siRNA mediated suppression of BAG3 production in neonatal rat ventricular cardiomyocytes (NRVCs) significantly elevates the level of Parkin, a key component of mitophagy. We found that both BAG3 and Parkin are recruited to depolarized mitochondria and promote mitophagy. Suppression of BAG3 in NRVCs significantly reduces autophagy flux and eliminates clearance of Tom20, an essential import receptor for mitochondria proteins, after induction of mitophagy. These observations suggest that BAG3 is critical for the maintenance of mitochondrial homeostasis under stress conditions, and disruptions in BAG3 expression impact cardiomyocyte function. J. Cell. Physiol. 232: 797–805, 2017. © 2016 Wiley Periodicals, Inc. BAG3 is critical for homeostasis of skeletal and cardiac tissues. Here we demonstrate that knockdown of BAG3 expression increased the level of Parkin, a key regulator of autophagy in neonatal rat ventricular cardiomyocytes. Furthermore, a decrease in the level of BAG3 promotes translocation of Parkin to the depolarized mitochondria and its degradation. BAG3 suppression significantly decreased the clearance of Tom20, an essential import receptor for mitochondria protein, after treatment with electron uncoupler, suggesting that mitophagy is impaired in the absence of BAG3. These observations ascribe a critical role for BAG3 in the maintenance of mitochondrial function under stress conditions.

  July 19, 2016   doi: 10.1002/jcp.25476   open full text
• SCD1 Alters Long‐Chain Fatty Acid (LCFA) Composition and Its Expression Is Directly Regulated by SREBP‐1 and PPARγ 1 in Dairy Goat Mammary Cells.
  Dawei Yao, Jun Luo, Qiuya He, Hengbo Shi, Jun Li, Hui Wang, Huifen Xu, Zhi Chen, Yongqing Yi, Juan J. Loor.
  Journal of Cellular Physiology. July 18, 2016
  Stearoyl‐CoA desaturase 1 (SCD1) is a key enzyme for the synthesis of the monounsaturated fatty acids (MUFA) palmitoleic acid and oleic acid. In non‐ruminant species, SCD1 expression is known to be tightly regulated by a variety of transcription factors. Although the role of SCD1 and the transcriptional regulatory mechanism by SREBP‐1 and PPARs in other species is clear, changes in lipid metabolism related to SCD1 and via the regulation of SREBP‐1 or PPARG1 in ruminant mammary tissue remain largely unknown. Here, we demonstrated that SCD1 expression in goat mammary tissue is higher during lactation than the dry period. Overexpression of SCD1 increased the intracellular MUFA content and lipid accumulation, whereas SCD1 silencing resulted in a significant decrease in oleic acid concentration and triacylglycerol (TAG) accumulation. The overexpression of SREBF1 in goat mammary epithelial cells (GMEC) enhanced SCD1 expression and its promoter activity, but that effect was abolished when SREBF1 was silenced. Furthermore, deletion of sterol regulatory element (SRE) and the nuclear factor (NF‐Y)‐binding sites within a −1713 to +65‐base pair region of the SCD1 promoter completely abolished SREBP‐1‐induced SCD1 transcription. Otherwise, PPARG1 overexpression also stimulated the expression of SCD1 and its transcriptional activity directly via a PPAR response element (PPRE) in the SCD1 promoter. Together, these results indicate that SCD1 could markedly affect the fatty acid composition and rate of TAG synthesis through direct regulation via SREBP‐1 and PPARG1, hence, underscoring an important role of the enzyme and this transcription regulator in controlling mammary gland lipid synthesis in the goat. J. Cell. Physiol. 232: 635–649, 2017. © 2016 Wiley Periodicals, Inc. The function of stearoyl‐CoA desaturase 1 gene in goat mammary cells. The regulatory mechanism of SCD1 by SREBP1 and PPARG in dairy goat.

  July 18, 2016   doi: 10.1002/jcp.25469   open full text
• Transcriptomic Analyses of Adipocyte Differentiation From Human Mesenchymal Stromal‐Cells (MSC).
  Antonio Casado‐Díaz, Jaouad Anter, Sören Müller, Peter Winter, José Manuel Quesada‐Gómez, Gabriel Dorado.
  Journal of Cellular Physiology. July 12, 2016
  Adipogenesis is a physiological process required for fat‐tissue development, mainly involved in regulating the organism energetic‐state. Abnormal distribution‐changes and dysfunctions in such tissue are associated to different pathologies. Adipocytes are generated from progenitor cells, via a complex differentiating process not yet well understood. Therefore, we investigated differential mRNA and miRNA expression patterns of human mesenchymal stromal‐cells (MSC) induced and not induced to differentiate into adipocytes by next (second)‐generation sequencing. A total of 2,866 differentially expressed genes (101 encoding miRNA) were identified, with 705 (46 encoding miRNA) being upregulated in adipogenesis. They were related to different pathways, including PPARG, lipid, carbohydrate and energy metabolism, redox, membrane‐organelle biosynthesis, and endocrine system. Downregulated genes were related to extracellular matrix and cell migration, proliferation, and differentiation. Analyses of mRNA‐miRNA interaction showed that repressed miRNA‐encoding genes can act downregulating PPARG‐related genes; mostly the PPARG activator (PPARGC1A). Induced miRNA‐encoding genes regulate downregulated genes related to TGFB1. These results shed new light to understand adipose‐tissue differentiation and physiology, increasing our knowledge about pathologies like obesity, type‐2 diabetes and osteoporosis. J. Cell. Physiol. 232: 771–784, 2017. © 2016 Wiley Periodicals, Inc. Adipogenesis transcriptomics was studied in mesenchymal stem‐cells (MSC) and 659 mRNA and 46 miRNA were upregulated in adipocyte differentiation. Metabolic, redox, and endocrinology process were upregulated in adipogenesis and Migration, proliferation, and differentiation to others cell types were downregulated. miRNA that can interact with PPARG and TGFB1 pathways were both downregulated and upregulated.

  July 12, 2016   doi: 10.1002/jcp.25472   open full text
• Parathyroid Hormone‐Related Protein Protects Osteoblastic Cells From Oxidative Stress by Activation of MKP1 Phosphatase.
  Juan A. Ardura, Sergio Portal‐Núñez, Irantzu Castelbón‐Calvo, Irene Martínez de Toda, Mónica De la Fuente, Pedro Esbrit.
  Journal of Cellular Physiology. July 12, 2016
  Oxidative damage is an important contributor to the morphological and functional changes in osteoporotic bone. Aging increases the levels of reactive oxygen species (ROS) that cause oxidative stress and induce osteoblast apoptosis. ROS modify several signaling responses, including mitogen‐activated protein kinase (MAPK) activation, related to cell survival. Both parathyroid hormone (PTH) and its bone counterpart, PTH‐related protein (PTHrP), can regulate MAPK activation by modulating MAPK phosphatase‐1 (MKP1). Thus, we hypothesized that PTHrP might protect osteoblasts from ROS‐induced apoptosis by targeting MKP1. In osteoblastic MC3T3‐E1 and MG‐63 cells, H2O2 triggered p38, JNK, ERK and p66Shc phosphorylation, and cell apoptosis. Meanwhile, PTHrP (1‐37) rapidly but transiently increased ERK and Akt phosphorylation without affecting p38, JNK, or p66Shc activation. H2O2‐induced p38 and ERK phosphorylation and apoptosis were both decreased by pre‐treatment with specific kinase inhibitors or PTHrP (1‐37) in both osteoblastic cell types. These dephosphorylating and prosurvival actions of PTHrP (1‐37) were prevented by a phosphatase inhibitor cocktail, the phosphatase MKP1 inhibitor sanguinarine or a MKP1 siRNA. PTHrP (1‐37) promptly enhanced MKP1 protein and gene expression and MKP1‐dependent catalase activity in osteoblastic cells. Furthermore, exposure to PTHrP (1‐37) adsorbed in an implanted hydroxyapatite‐based ceramic into a tibial defect in aging rats increased MKP1 and catalase gene expression in the healing bone area. Our findings demonstrate that PTHrP counteracts the pro‐apoptotic actions of ROS by a mechanism dependent on MKP1‐induced dephosphorylation of MAPKs in osteoblasts. J. Cell. Physiol. 232: 785–796, 2017. © 2016 Wiley Periodicals, Inc. PTHrP counteracts the pro‐apoptotic actions of ROS by a mechanism dependent on MKP1 dephosphorylation of MAP kinases and decreasing reactive oxygen species accumulation.

  July 12, 2016   doi: 10.1002/jcp.25473   open full text
• Generation of Human Lens Epithelial‐Like Cells From Patient‐Specific Induced Pluripotent Stem Cells.
  Dan Li, Xiaodi Qiu, Jin Yang, Tianjin Liu, Yi Luo, Yi Lu.
  Journal of Cellular Physiology. July 11, 2016
  Cataractogenesis begins from the dynamic lens epithelial cells (LECs) and adjacent fiber cells. LECs derived from cell lines cannot maintain the crystalline expression as the primary LECs. The current study aimed to efficiently generate large numbers of human LECs from patient‐specific induced pluripotent stem cells (iPSCs). Anterior lens capsules were collected from cataract surgery and were used to culture primary hLECs. iPSCs were induced from these primary hLECs by lentiviral transduction of Oct4, Sox2, Klf4, and c‐Myc. Then, the generated iPSCs were re‐differentiated into hLECs by the 3‐step addition of defined factor combinations (Noggin, BMP4/7, bFGF, and EGF) modified from an established method. During the re‐differentiation process, colonies of interest were isolated using a glass picking tool and cloning cylinders based on the colony morphology. After two steps of isolation, populations of LEC‐like cells (LLCs) were generated and identified by the expression of lens marker genes by qPCR, western blot and immunofluorescence staining. The study introduced a modified protocol to isolate LLCs from iPSCs by defined factors in a short time frame. This technique could be useful for mechanistic studies of lens‐related diseases. J. Cell. Physiol. 231: 2555–2562, 2016. © 2016 Wiley Periodicals, Inc. The study introduced a modified protocol to isolate lens epithelial like cells from cataract patient‐derived iPSCs by defined factors in a short time frame.

  July 11, 2016   doi: 10.1002/jcp.25374   open full text
• Intricate Functions of Matrix Metalloproteinases in Physiological and Pathological Conditions.
  Rahul Mittal, Amit P. Patel, Luca H. Debs, Desiree Nguyen, Kunal Patel, M'hamed Grati, Jeenu Mittal, Denise Yan, Prem Chapagain, Xue Zhong Liu.
  Journal of Cellular Physiology. July 11, 2016
  Matrix metalloproteinases (MMPs) are a diverse group of proteolytic enzymes and play an important role in the degradation and remodeling of the extracellular matrix (ECM). In normal physiological conditions, MMPs are usually minimally expressed. Despite their low expression, MMPs have been implicated in many cellular processes ranging from embryological development to apoptosis. The activity of MMPs is controlled at three different stages: (1) transcription; (2) zymogen activation; and (3) inhibition of active forms by tissue inhibitor metalloproteinases (TIMPs). They can collectively degrade any component of ECM and basement membrane, and their excessive activity has been linked to numerous pathologies mainly including, but not limited to, tumor invasion and metastasis. The lack of information about several MMPs and the steady stream of new discoveries suggest that there is much more to be studied in this field. In particular, there is a need for controlling their expression in disease states. Various studies over the past 30 years have found that each MMP has a specific mode of activation, action, and inhibition. Drugs specifically targeting individual MMPs could revolutionize the treatment of a great number of health conditions and tremendously reduce their burden. In this review article, we have summarized the recent advances in understanding the role of MMPs in physiological and pathological conditions. J. Cell. Physiol. 231: 2599–2621, 2016. © 2016 Wiley Periodicals, Inc. Ribbon diagram of the full structure of MMP‐9 created with Swiss‐model and rendered with Pymol showing pro and catalytic domains.

  July 11, 2016   doi: 10.1002/jcp.25430   open full text
• Cholesterol and Its Derivatives Reversibly Inhibit Proteinase K.
  Namrata Singh, Debasish Bhattacharyya.
  Journal of Cellular Physiology. July 09, 2016
  Microorganisms express a variety of proteases that degrade many proteins of the host body and subvert host immune response. While elucidating the mechanism/s of an immune stimulatory drug that contains bile lipid, regulation of proteolytic activity was investigated. The drug and bile lipids both stabilize Proteinase K, an aggressive protease of fungal origin against auto‐digestion. Among the components of bile lipids, only cholesterol and its derivatives stabilize the enzyme. Biophysical evidences such as scattering of light, intrinsic and extrinsic fluorescence emission spectra, circular dichroism spectra, atomic force microscopy, and transmission electron microscopy images indicated that cholesterol and its derivatives interact with Proteinase K. Inhibition kinetics using esterolysis of ATEE revealed non‐competitive inhibition by cholesterol. Surface Plasmon Resonance and mass spectrometric analysis indicated 1:1 stoichiometry of binding and with dissociation constant in the μM range. Further, the presence of four cholesterol recognition amino acid consensus motifs (CRAC motifs I–IV) was identified in Proteinase K. Bioinformatics analysis revealed that the whole stretch of cholesterol interacts very well with the hydrophobic groove of motif II only among the four CRAC motifs. Variation of cholesterol content of HepG2 human liver carcinoma cells showed positive correlation with binding of fluorescence tagged Proteinase K. Under these conditions, binding of Proteinase K to the cells did not affect their morphology, viability and growth kinetics. Cell bound Proteinase K could be released by an excess of its substrate, thereby restoring reversibly its proteolytic activity. J. Cell. Physiol. 232: 596–609, 2017. © 2016 Wiley Periodicals, Inc. Cholesterol and its derivatives reversibly inactivates Proteinase K, an aggressive protease of fungal origin against its auto‐digestion. In ex‐vivo studies, confocal images indicated that membrane cholesterol reversibly binds with the enzyme.

  July 09, 2016   doi: 10.1002/jcp.25457   open full text
• Cartilage‐Specific and Cre‐Dependent Nkx3.2 Overexpression In Vivo Causes Skeletal Dwarfism by Delaying Cartilage Hypertrophy.
  Da‐Un Jeong, Je‐Yong Choi, Dae‐Won Kim.
  Journal of Cellular Physiology. July 05, 2016
  Nkx3.2, the vertebrate homologue of Drosophila bagpipe, has been implicated as playing a role in chondrogenic differentiation. In brief, Nkx3.2 is initially expressed in chondrocyte precursor cells and later during cartilage maturation, its expression is diminished in hypertrophic chondrocytes. In addition to Nkx3.2 expression analyses, previous studies using ex vivo chick embryo cultures and in vitro cell cultures have suggested that Nkx3.2 can suppress chondrocyte hypertrophy. However, it has never been demonstrated that Nkx3.2 functions in regulating chondrocyte hypertrophy during cartilage development in vivo. Here, we show that cartilage‐specific and Cre‐dependent Nkx3.2 overexpression in mice results in significant postnatal dwarfism in endochondral skeletons, while intramembranous bones remain unaltered. Further, we observed significant delays in cartilage hypertrophy in conditional transgenic ciTg‐Nkx3.2 mice. Together, these findings confirm that Nkx3.2 is capable of controlling hypertrophic maturation of cartilage in vivo, and this regulation plays a significant role in endochondral ossification and longitudinal bone growth. J. Cell. Physiol. 9999: 1–13, 2016. © 2016 Wiley Periodicals, Inc. Cartilage‐specific and Cre‐dependent Nkx3.2 overexpression in mice results in remarkable postnatal dwarfism due to notable delays in cartilage hypertrophy. These findings indicate Nkx3.2 is capable of controlling hypertrophic maturation of cartilage in vivo, and this regulation plays a significant role in endochondral ossification and longitudinal bone growth.

  July 05, 2016   doi: 10.1002/jcp.25446   open full text
• MicroRNA‐30b Suppresses Epithelial‐Mesenchymal Transition and Metastasis of Hepatoma Cells.
  Xiaolin Sun, Shuhua Zhao, Huanan Li, Hanwen Chang, Zhen Huang, Zhi Ding, Lei Dong, Jiangning Chen, Yuhui Zang, Junfeng Zhang.
  Journal of Cellular Physiology. June 30, 2016
  Epithelial–mesenchymal transition (EMT) is critical for induction of invasiveness and metastasis in HCC. Growing evidence indicates that upregulation of Snail, the major EMT inducer, significantly correlates with the metastasis and poor prognosis of HCC. Here, we investigate the underlying mechanism of miR‐30b in suppressing metastasis of hepatoma cells by targeting Snail. In this study, we found that miR‐30b was significantly downregulated and negatively associated with Snail production in HCC cell lines with higher metastatic potentials. Gain‐ and loss‐of‐function studies revealed that miR‐30b could dramatically inhibit in vitro HCC cell migration and invasion. In vivo orthotopic liver xenograft model further demonstrated that stable over‐expression of miR‐30b significantly repressed the local invasion and lung metastasis of hepatoma cells. Meanwhile, the restoration of miR‐30b expression suppressed the distant colonization of hepatoma cells. Both gain‐ and loss‐of‐function studies showed that miR‐30b suppressed the EMT of hepatoma cells as indicated by the morphology changes and deregulation of epithelial and mesenchymal markers. Using RNAi, we further investigated the role of Snail in HCC cell EMT and demonstrated that knockdown of Snail significantly inhibited the EMT and cancer cell metastasis. Additionally, miR‐30b exhibited inhibitory effects on HCC cell proliferation in vitro and in vivo. In conclusion, our findings highlight the significance of miR‐30b downregulation in HCC tumor metastasis and invasiveness, and implicate a new potential therapeutic target for HCC metastasis. J. Cell. Physiol. 232: 625–634, 2017. © 2016 Wiley Periodicals, Inc. miR‐30b was significantly downregulated and negatively associated with Snai1 production in HCC cell lines with higher metastatic potentials. miR‐30b exerts its suppressive function on HCC metastasis by inhibiting the Snail‐driven EMT.

  June 30, 2016   doi: 10.1002/jcp.25466   open full text
• DGKζ Downregulation Enhances Osteoclast Differentiation and Bone Resorption Activity Under Inflammatory Conditions.
  Kiyoshi Iwazaki, Toshiaki Tanaka, Yasukazu Hozumi, Masashi Okada, Rieko Tsuchiya, Ken Iseki, Matthew K. Topham, Kaneyuki Kawamae, Michiaki Takagi, Kaoru Goto.
  Journal of Cellular Physiology. June 30, 2016
  Bone homeostasis is maintained by a balance between resorption of the bone matrix and its replacement by new bone. Osteoclasts play a crucially important role in bone metabolism. They are responsible for bone resorption under pathophysiological conditions. Differentiation of these cells, which are derived from bone marrow cells, depends on receptor activator of NF‐κB ligand (RANKL). RANKL‐induced osteoclastogenesis is regulated by the phosphoinositide (PI) signaling pathway, in which diacylglycerol (DG) serves as a second messenger in signal transduction. In this study, we examined the functional implications of DG kinase (DGK), an enzyme family responsible for DG metabolism, for osteoclast differentiation and activity. Of DGKs, DGKζ is most abundantly expressed in osteoclast precursors such as bone marrow‐derived monocytes/macrophages. During osteoclast differentiation from precursor cells, DGKζ is downregulated at the protein level. In this regard, we found that DGKζ deletion enhances osteoclast differentiation and bone resorption activity under inflammatory conditions in an animal model of osteolysis. Furthermore, DGKζ deficiency upregulates RANKL expression in response to TNFα stimulation. Collectively, results suggest that DGKζ is silent under normal conditions, but it serves as a negative regulator in osteoclast function under inflammatory conditions. Downregulation of DGKζ might be one factor predisposing a person to osteolytic bone destruction in pathological conditions. J. Cell. Physiol. 232: 617–624, 2017. © 2016 Wiley Periodicals, Inc. The DGK family is involved in cellular signal transduction. Of DGKs, DGKζ serves as a negative regulator in osteoclast function. Downregulation of DGKζ enhances bone destruction in pathological conditions.

  June 30, 2016   doi: 10.1002/jcp.25461   open full text
• FGF23 Neutralizing Antibody Ameliorates Hypophosphatemia and Impaired FGF Receptor Signaling in Kidneys of HMWFGF2 Transgenic Mice.
  E. Du, L. Xiao, M.M. Hurley.
  Journal of Cellular Physiology. June 30, 2016
  High molecular weight FGF2 transgenic mice (HMWTg) phenocopy the Hyp mouse, homolog of human X‐linked hypophosphatemic rickets with phosphate wasting and abnormal fibroblast growth factor (FGF23), fibroblast growth factor receptor (FGFR), Klotho and mitogen activated protein kinases (MAPK) signaling in kidney. In this study, we assessed whether short‐term (24 h) in vivo administration of FGF23 neutralizing antibody (FGF23Ab) could rescue hypophosphatemia and impaired FGFR signaling in kidneys of HMWTg male mice. Bone mineral density and bone mineral content in 1‐month‐old HMWTg mice were significantly reduced compared with Control/VectorTg mice. Serum FGF23 was significantly increased in HMWTg compared with VectorTg. Serum phosphate was significantly reduced in HMWTg and was rescued by FGF23Ab. Serum parathyroid hormone (PTH) was significantly increased in HMWTg but was not reduced by FGF23Ab. 1, 25(OH)2 D was inappropriately normal in serum of HMWTg and was significantly increased in both Vector and HMWTg by FGF23Ab. Analysis of HMWTg kidneys revealed significantly increased mRNA expression of the FGF23 co‐receptor Klotho, transcription factor mRNAs for early growth response‐1 transcription factor (Egr‐1), and c‐fos were all significantly decreased by FGF23Ab. A significant reduction in the phosphate transporter Npt2a mRNA was also observed in HMWTg kidneys, which was increased by FGF23Ab. FGF23Ab reduced p‐FGFR1, p‐FGFR3, KLOTHO, p‐ERK1/2, C‐FOS, and increased NPT2A protein in HMWTg kidneys. We conclude that FGF23 blockade rescued hypophosphatemia by regulating FGF23/FGFR downstream signaling in HMWTg kidneys. Furthermore, HMWFGF2 isoforms regulate PTH expression independent of FGF23/FGFR signaling. J. Cell. Physiol. 232: 610–616, 2017. © 2016 Wiley Periodicals, Inc. A schematic model for the role of HMWFGF2 in the regulation of Pi by FGF23 signaling. Overexpression of HMWFGF2 in osteoblast linage cells induced excess FGF23 in blood. Elevated FGF23 can bind to its co‐receptors Fgfr1c/Fgfr3c and Klotho on kidney to activate ERK1/2 signaling pathway in distal tubule, this signaling was transferred to Npt2a in proximal tubules, resulting in Pi wasting. Blockade of FGF23 with FGF23Ab can rescue hypophosphatemia and impaired FGFR signaling in kidneys of HMWTg mice. Activated ERK1/2 can lead to hypophosphatemia by suppression of Cyp27b1 and induction of Cyp24.

  June 30, 2016   doi: 10.1002/jcp.25458   open full text
• Apigenin Reduces Survival of Choriocarcinoma Cells by Inducing Apoptosis via the PI3K/AKT and ERK1/2 MAPK Pathways.
  Whasun Lim, Sunwoo Park, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. June 29, 2016
  Apigenin is a flavonoid found in parsley, onions, oranges, tea, chamomile, wheat, and sprouts. It has a variety of biological properties including anti‐oxidant, anti‐mutagenic, anti‐carcinogenic, anti‐inflammatory, anti‐proliferative, and anti‐spasmodic effects. Based on epidemiological and case‐control studies, apigenin is regarded as a novel chemotherapeutic agent against various cancer types. However, little is known about the effects of apigenin on choriocarcinoma cells. Therefore, we investigated the anti‐cancer effects of apigenin on choriocarcinoma cells (JAR and JEG3) in the present study. Apigenin reduced viability and migratory properties, increased apoptosis, and suppressed mitochondrial membrane potential in both the JAR and JEG3 cells. In addition, apigenin predominantly decreased phosphorylation of AKT, P70RSK, and S6 whereas the phosphorylation of ERK1/2 and P90RSK was increased by apigenin treatment of JAR and JEG3 cells in a dose‐dependent manner. Moreover, treatment of JAR and JEG3 cells with both apigenin and pharmacological inhibitors of PI3K/AKT (LY294002) and ERK1/2 (U0126) revealed synergistic anti‐proliferative effects. Collectively, these results indicated that the apigenin is an invaluable chemopreventive agent that inhibits progression and metastasis of choriocarcinoma cells through regulation of PI3K/AKT and ERK1/2 MAPK signal transduction mechanism. J. Cell. Physiol. 231: 2690–2699, 2016. © 2016 Wiley Periodicals, Inc. Results of this study provide the first evidence that apigenin acts as chemotherapeutic agent on progression and metastasis of choriocarcinoma cells through regulating PI3K/AKT and ERK1/2 MAPK signal transduction cascades.

  June 29, 2016   doi: 10.1002/jcp.25372   open full text
• Affinity Selection of FGF2‐Binding Heparan Sulfates for Ex Vivo Expansion of Human Mesenchymal Stem Cells.
  Sampath Jeewantha Wijesinghe, Ling Ling, Sadasivam Murali, Yeong Hui Qing, Simon F.R. Hinkley, Susan M. Carnachan, Tracey J. Bell, Kunchithapadam Swaminathan, James H. Hui, Andre J. van Wijnen, Victor Nurcombe, Simon M. Cool.
  Journal of Cellular Physiology. June 28, 2016
  The future of human mesenchymal stem cells (hMSCs) as a successful cell therapy relies on bioprocessing strategies to improve the scalability of these cells without compromising their therapeutic ability. The culture‐expansion of hMSCs can be enhanced by supplementation with growth factors, particularly fibroblast growth factor 2 (FGF2). The biological activity of FGF2 is controlled through interactions with heparan sulfate (HS) that facilitates ligand‐receptor complex formation. We previously reported on an FGF2‐interacting HS variant (termed HS2) isolated from embryonic tissue by anionic exchange chromatography that increased the proliferation and potency of hMSCs. Here, we detail the isolation of an FGF2 affinity‐purified HS variant (HS8) using a scalable platform technology previously employed to generate HS variants with increased affinity for BMP‐2 or VEGF165. This process used a peptide sequence derived from the heparin‐binding domain of FGF2 as a substrate to affinity‐isolate HS8 from a commercially available source of porcine mucosal HS. Our data show that HS8 binds to FGF2 with higher affinity than to FGF1, FGF7, BMP2, PDGF‐BB, or VEGF165. Also, HS8 protects FGF2 from thermal destabilization and increases FGF signaling and hMSC proliferation through FGF receptor 1. Long‐term supplementation of cultures with HS8 increased both hMSC numbers and their colony‐forming efficiency without adversely affecting the expression of hMSC‐related cell surface antigens. This strategy further exemplifies the utility of affinity‐purifying HS variants against particular ligands important to the stem cell microenvironment and advocates for their addition as adjuvants for the culture‐expansion of hMSCs destined for cellular therapy. J. Cell. Physiol. 232: 566–575, 2017. © 2016 Wiley Periodicals, Inc. We here report a novel strategy for ex vivo expansion of human mesenchymal stem cells that replies on supplementation of growth factor‐binding heparan sulfates, instead of the growth factor itself into the culture. The advantage of such approach is to avoid the reported adverse effect of FGF2 on MSCs upon long‐term exogenous supplementation, and more importantly, the quality of hMSCs expanded is not compromised after long‐term culture. The work is of particular importance given the wide therapeutic use of hMSCs and the key role of FGF2 in the technology of hMSC scaling‐up.

  June 28, 2016   doi: 10.1002/jcp.25454   open full text
• Post‐Transcriptional Regulation of the Human Mu‐Opioid Receptor (MOR) by Morphine‐Induced RNA Binding Proteins hnRNP K and PCBP1.
  Kyu Young Song, Hack Sun Choi, Ping‐Yee Law, Li‐Na Wei, Horace H. Loh.
  Journal of Cellular Physiology. June 24, 2016
  Expression of the mu‐opioid receptor (MOR) protein is controlled by extensive transcriptional and post‐transcriptional processing. MOR gene expression has previously been shown to be altered by a post‐transcriptional mechanism involving the MOR mRNA untranslated region (UTR). Here, we demonstrate for the first time the role of heterogeneous nuclear ribonucleic acids (hnRNA)‐binding protein (hnRNP) K and poly(C)‐binding protein 1 (PCBP1) as post‐transcriptional inducers in MOR gene regulation. In the absence of morphine, a significant level of MOR mRNA is sustained in its resting state and partitions in the translationally inactive polysomal fraction. Morphine stimulation activates the downstream targets hnRNP K and PCPB1 and induces partitioning of the MOR mRNA to the translationally active fraction. Using reporter and ligand binding assays, as well as RNA EMSA, we reveal potential RNP binding sites located in the 5′‐untranslated region of human MOR mRNA. In addition, we also found that morphine‐induced RNPs could regulate MOR expression. Our results establish the role of hnRNP K and PCPB1 in the translational control of morphine‐induced MOR expression in human neuroblastoma (NMB) cells as well as cells stably expressing MOR (NMB1). J. Cell. Physiol. 232: 576–584, 2017. © 2016 Wiley Periodicals, Inc. During stimulation, the vast majority of MOR mRNA partitions to the polysomal fraction for synthesis of MOR proteins. These effects are more evident in NMB1 cells, which stably express MOR.

  June 24, 2016   doi: 10.1002/jcp.25455   open full text
• Inhibition of mTOR Signaling Pathway Delays Follicle Formation in Mice.
  Jing Zhang, Wenwen Liu, Xinhui Sun, Feifei Kong, Ye Zhu, Yue Lei, Youqiang Su, Yiping Su, Jing Li.
  Journal of Cellular Physiology. June 24, 2016
  In mammalian ovaries, follicle assembly requires proper germ cell cyst breakdown and the invasion of somatic cells to encapsulate individual oocytes. Abnormalities in this process lead to a number of pathologies such as premature ovarian failure and infertility. As a conserved pathway regulating cell growth and metabolism in response to growth factors and nutrients, the roles of mTOR signaling in follicular development have been extensively studied in recent years. However, its functions during follicle formation remain unknown. In this study, the expression of p‐rpS6 (phospho‐ribosomal proteinS6), a downstream marker of mTORC1, showed dynamic changes in perinatal ovaries. When E18.5 ovaries, which mainly contained germ cell nests, were incubated with the mTOR inhibitors Rapamycin and Torin1 for 24 h, follicle assembly was delayed with differential somatic cell invasion into germ cell cyst among the groups. After transplanting treated or untreated ovaries into kidney capsules of recipient ovariectomized mice, follicular development was blocked in treated ovaries, as shown by fewer antral follicles and a higher proportion of primordial follicles. Further studies showed a significant decrease in somatic cell proliferation and the expression of marker genes related to follicular development (Kitl, Kit, Gdf9, Bmp15, Zp3, and Amhr2) in treated ovaries. Moreover, the addition of KITL, a growth factor that is mainly produced by pregranulosa cells during germ cell nest breakdown, rescued the extension of follicle formation induced by mTOR inhibitors. These results suggest that KITL functions downstream of mTOR in somatic cells to regulate their communication with oocytes during follicle formation. J. Cell. Physiol. 232: 585–595, 2017. © 2016 Wiley Periodicals, Inc. In mammalian ovaries, follicle assembly requires proper germ cell cyst breakdown and the invasion of somatic cells to encapsulate individual oocytes. In the study, follicle formation was delayed after inhibiting the mTOR signaling pathway by mTOR inhibitors. Moreover, KITL functions downstream of mTOR in somatic cells to regulate their communication with oocytes during follicle formation.

  June 24, 2016   doi: 10.1002/jcp.25456   open full text
• T11TS Treatment Augments Apoptosis of Glioma Associated Brain Endothelial Cells, Hint Toward Anti‐Angiogenic Action in Glioma.
  Debanjan Bhattacharya, Manoj Kumar Singh, Suhnrita Chaudhuri, Ankur Datta, Swapna Chaudhuri.
  Journal of Cellular Physiology. June 21, 2016
  Malignant glioma continues to be a clinical challenge with an urgent need for developing curative therapeutic intervention. Apoptosis induction in tumor‐associated endothelial cells represent a central mechanism that counteracts angiogenesis in glioma and other solid tumors. We previously demonstrated that intraperitoneal administration of sheep erythrocyte membrane glycopeptide T11‐target structure (T11TS) in rodent glioma model inhibits PI3K/Akt pathway and Raf/MEK/ERK signaling in glioma‐associated brain endothelial cells. In the present study, we investigated whether T11TS treatment influence apoptosis signaling in vivo in glioma‐associated brain endothelial cells. Annexin‐V/PI staining showed that T11TS treatment in glioma‐induced rats increases apoptosis of glioma‐associated endothelial cells within glioma milieu compared to brain endothelial cells in glioma induced and control groups. Flowcytometric JC‐1 assay revealed that T11TS administration triggers loss of mitochondrial membrane potential in glioma‐associated brain endothelial cells. Flowcytometry, immunoblotting, and in situ immunofluoresecnt imaging were employed to investigate the effect of T11TS on apoptotic regulatory proteins in brain endothelial cells. T11TS treatment‐upmodulated expression of p53, Bax, Fas, FasL, and FADD in glioma associated endothelial cells and downregulated Bcl‐2 protein. T11TS therapy induced cytochrome‐c release into cytosol, activated caspase −9, 8, 3, and cleaved Bid in glioma associated brain endothelial cells. The study demonstrates that T11TS induces apoptosis in glioma‐associated brain endothelial cells via p53 accumulation and activation of intrinsic as well as Fas‐dependent extrinsic pathway. The pro‐apoptotic action of T11TS on glioma‐associated endothelial cells provides crucial insight into how T11TS exerts its anti‐angiogenic function in glioma. J. Cell. Physiol. 232: 526–539, 2017. © 2016 Wiley Periodicals, Inc. T11TS induced apoptosis of glioma‐associated brain endothelial cells and its effect on glioma angiogenesis.

  June 21, 2016   doi: 10.1002/jcp.25447   open full text
• In Vitro and In Vivo Differentiation of Progenitor Stem Cells Obtained After Mechanical Digestion of Human Dental Pulp.
  Manuela Monti, Antonio Graziano, Silvana Rizzo, Cesare Perotti, Claudia Del Fante, Riccardo d'Aquino, Carlo Alberto Redi, Ruggero Rodriguez y Baena.
  Journal of Cellular Physiology. June 21, 2016
  Human population is facing a revolutionary change in the demographic structure with an increasing number of elderly people requiring an unmet need to ensure a smooth aging process and dental care is certainly an important aspect that has to be considered. To date, dentistry has been conservative and the need of transferring the scientific models of regenerative dentistry into clinical practice is becoming a necessity. The aim of this study was to characterize the differentiation commitment (in vitro) and the clinical grafting ability (in vivo) of a population of progenitor stem cells obtained after mechanical digestion of dental pulp with an innovative system recently developed. This approach was successfully used in previous studies to obtain a clinical‐grade ready to use dental pulp fragments that could be grafted in autologous tissues to obtain bone. We are thus showing that micro grafts resulting from mechanical digestion contain stem cells with a mesenchymal phenotype, able to differentiate toward different cell types and to generate new bone in patients. We are providing data for the establishment of standardized and routinely oral surgery approaches, having outlined the cellular properties of human stem cells obtained from the dental pulp. This method can represent a valid tool for both regenerative medicine and tissue engineering purposes not only applicable to the cranio‐maxillofacial region but, likely, to different bone pathologies for a fastening and healing recovering of patients. J. Cell. Physiol. 232: 548–555, 2017. © 2016 Wiley Periodicals, Inc. This paper presents a new approach to obtain a clinical‐grade ready to use dental pulp fragments that could be grafted in autologous tissues to obtain bone. Micro‐grafts resulting from mechanical digestion contain stem cells with a mesenchymal phenotype, able to differentiate toward different cell types and to generate new bone in patients.

  June 21, 2016   doi: 10.1002/jcp.25452   open full text
• Browning of White Fat: Novel Insight Into Factors, Mechanisms, and Therapeutics.
  Nevena Jeremic, Pankaj Chaturvedi, Suresh C. Tyagi.
  Journal of Cellular Physiology. June 21, 2016
  What is more interesting about brown adipose tissue (BAT) is its ability to provide thermogenesis, protection against obesity by clearing triglycerides, releasing batokines, and mitigating insulin resistance. White adipose tissue (WAT) on the other hand stores excess energy and secretes some endocrine factors like leptin for regulating satiety. For the last decade there has been an increasing interest in the browning of fat keeping in view its beneficial effects on metabolic disorders and protection in the form of perivascular fat. Obesity is one such metabolic disorder that leads to significant morbidity and mortality from obesity‐related disorders such as type 2 diabetes mellitus (T2D) and cardiovascular disease risk. Browning of white fat paves the way to restrict obesity and obesity related disorders. Although exercise has been the most common factor for fat browning; however, there are other factors that involve: (1) beta aminoisobutyric acid (BAIBA); (2) gamma amino butyric acid (GABA); (3) PPARɣ agonists; (4) JAK inhibition; and (5) IRISIN. In this review, we propose two novel factors musclin and TFAM for fat browning. Musclin a myokine released from muscles during exercise activates PPARɣ which induces browning of WAT that has beneficial metabolic and cardiac effects. TFAM is a transcription factor that induces mitochondrial biogenesis. Since BAT is rich in mitochondria, higher expression of TFAM in WAT or TFAM treatment in WAT cells can induce browning of WAT. We propose that fat browning can be used as a therapeutic tool for metabolic disorders and cardiovascular diseases. J. Cell. Physiol. 232: 61–68, 2017. © 2016 Wiley Periodicals, Inc. What is more interesting about brown adipose tissue (BAT) is its ability to provide thermogenesis, protection against obesity by clearing triglycerides, and mitigating insulin resistance. This review describes the important therapeutic aspects of fat browning through factors like GABA, BAIBA, PPARϒ, IRISIN, musclin, and TFAM.

  June 21, 2016   doi: 10.1002/jcp.25450   open full text
• Alternative Start Codon Connects eIF5A to Mitochondria.
  Karina Danielle Pereira, Letícia Tamborlin, Letícia Meneguello, André Ricardo Gomes de Proença, Isadora Cristina de Paula Andrade Almeida, Rogério Ferreira Lourenço, Augusto Ducati Luchessi.
  Journal of Cellular Physiology. June 21, 2016
  Eukaryotic translation initiation factor 5A (eIF5A), a protein containing the amino acid residue hypusine required for its activity, is involved in a number of physiological and pathological cellular processes. In humans, several EIF5A1 transcript variants encode the canonical eIF5A1 isoform B, whereas the hitherto uncharacterized variant A is expected to code for a hypothetical eIF5A1 isoform, referred to as isoform A, which has an additional N‐terminal extension. Herein, we validate the existence of eIF5A1 isoform A and its production from transcript variant A. In fact, variant A was shown to encode both eIF5A1 isoforms A and B. Mutagenic assays revealed different efficiencies in the start codons present in variant A, contributing to the production of isoform B at higher levels than isoform A. Immunoblotting and mass spectrometric analyses showed that isoform A can undergo hypusination and acetylation at specific lysine residues, as observed for isoform B. Examination of the N‐terminal extension suggested that it might confer mitochondrial targeting. Correspondingly, we found that isoform A, but not isoform B, co‐purified with mitochondria when the proteins were overproduced. These findings suggest that eIF5A1 isoform A has a role in mitochondrial function. J. Cell. Physiol. 231: 2682–2689, 2016. © 2016 Wiley Periodicals, Inc. Eukaryotic translation initiation factor 5A (eIF5A), a protein containing the amino acid residue hypusine required for its activity, is involved in a number of physiological and pathological cellular processes. Examination of the N‐terminal extension of the isoform A suggested that it might confer mitochondrial targeting. Correspondingly, we found that isoform A, but not isoform B, co‐purified with mitochondria when the proteins were overproduced. These findings suggest that eIF5A1 isoform A has a role in mitochondrial function.

  June 21, 2016   doi: 10.1002/jcp.25370   open full text
• Targeted Modification of Mitochondrial ROS Production Converts High Glucose‐Induced Cytotoxicity to Cytoprotection: Effects on Anesthetic Preconditioning.
  Filip Sedlic, Maria Y. Muravyeva, Ana Sepac, Marija Sedlic, Anna Marie Williams, Meiying Yang, Xiaowen Bai, Zeljko J. Bosnjak.
  Journal of Cellular Physiology. June 21, 2016
  Contradictory reports on the effects of diabetes and hyperglycemia on myocardial infarction range from cytotoxicity to cytoprotection. The study was designed to investigate acute effects of high glucose‐driven changes in mitochondrial metabolism and osmolarity on adaptive mechanisms and resistance to oxidative stress of isolated rat cardiomyocytes. We examined the effects of high glucose on several parameters of mitochondrial bioenergetics, including changes in oxygen consumption, mitochondrial membrane potential, and NAD(P)H fluorometry. Effects of high glucose on the endogenous cytoprotective mechanisms elicited by anesthetic preconditioning (APC) and the mediators of cell injury were also tested. These experiments included real‐time measurements of reactive oxygen species (ROS) production and mitochondrial permeability transition pore (mPTP) opening in single cells by laser scanning fluorescence confocal microscopy, and cell survival assay. High glucose rapidly enhanced mitochondrial energy metabolism, observed by increase in NAD(P)H fluorescence intensity, oxygen consumption, and mitochondrial membrane potential. This substantially elevated production of ROS, accelerated opening of the mPTP, and decreased survival of cells exposed to oxidative stress. Abrogation of high glucose‐induced mitochondrial hyperpolarization with 2,4 dinitrophenol (DNP) significantly, but not completely, attenuated ROS production to a level similar to hyperosmotic mannitol control. DNP treatment reversed high glucose‐induced cytotoxicity to cytoprotection. Hyperosmotic mannitol treatment also induced cytoprotection. High glucose abrogated APC‐induced mitochondrial depolarization, delay in mPTP opening and cytoprotection. In conclusion, high glucose‐induced mitochondrial hyperpolarization abolishes APC and augments cell injury. Attenuation of high glucose‐induced ROS production by eliminating mitochondrial hyperpolarization protects cardiomyocytes. J. Cell. Physiol. 232: 216–224, 2017. © 2016 Wiley Periodicals, Inc. High glucose‐induced excessive ROS production driven by combined effects of high mitochondrial membrane potential and hyperosmolarity acutely exacerbates cardiomyocyte injury. Moderate ROS production driven only by hyperosmolarity, with abrogated mitochondrial hyperpolarization, triggers adaptive response and protects cardiomyocytes. High glucose‐induced increase in ΔΨm overrides APC‐induced mitochondrial depolarization and blocks cytoprotection.

  June 21, 2016   doi: 10.1002/jcp.25413   open full text
• Leucine‐Rich Amelogenin Peptide (LRAP) Uptake by Cementoblast Requires Flotillin‐1 Mediated Endocytosis.
  Luciane Martins, Adriana Franco Paes Leme, Kamila Rosamilia Kantovitz, Em nome de Luciane Martins, Enilson Antonio Sallum, Márcio Zaffalon Casati, Francisco Humberto Nociti.
  Journal of Cellular Physiology. June 20, 2016
  Basic, pre‐clinical, and clinical studies have documented the potential of amelogenin, and its variants, to affect cell response and tissue regeneration. However, the mechanisms are unclear. Thus, the aim of the present study was to identify, in cementoblasts, novel binding partners for an alternatively spliced amelogenin form (Leucine‐Rich Amelogenin Peptide—LRAP), which is supposed to act as a signaling molecule in epithelial–mesenchymal interactions. LRAP‐binding protein complexes from immortalized murine cementoblasts (OCCM‐30) were achieved by capture affinity assay (GST pull down) and proteins present in these complexes were identified by mass spectrometry and immunoblotting. Flotillin‐1, which functions as a platform for signal transduction, vesicle trafficking, endocytosis, and exocytosis, was identified and confirmed by co‐precipitation and co‐localization assays as a protein‐binding partner for LRAP in OCCM‐30 cells. In addition, we found that exogenously added GST‐LRAP recombinant protein was internalized by OCCM‐30 cells, predominantly localized in the perinuclear region and, that inhibition of flotillin1‐dependent functions by small interference RNA (siRNA) methodology significantly affected LRAP uptake and its biological properties on OCCM‐30 cells, including LRAP effect on the expression of genes encoding osteocalcin (Ocn), bone sialoprotein (Bsp), and runt‐related transcription factor 2 (RunX2). In conclusion, LRAP uptake by cementoblast involves flotillin‐assisted endocytosis, which suggests an involvement of LRAP in lipid‐raft‐dependent signaling pathways which are mediated by flotillin‐1. J. Cell. Physiol. 232: 556–565, 2017. © 2016 Wiley Periodicals, Inc. Flotillin‐1, a lipid raft microdomain‐associated protein involved in intracellular trafficking pathways, endocytosis, and signal transduction, was identified as a protein‐binding partner for Leucine‐Rich Amelogenin Peptide (LRAP) impacting on LRAP‐mediated expression of key genes involved in cementoblastic differentiation.

  June 20, 2016   doi: 10.1002/jcp.25453   open full text
• Ethanol Extract of Cissus quadrangularis Enhances Osteoblast Differentiation and Mineralization of Murine Pre‐Osteoblastic MC3T3‐E1 Cells.
  Raazia Tasadduq, Jonathan Gordon, Khalid A. Al‐Ghanim, Jane B. Lian, Andre J. Van Wijnen, Janet L. Stein, Gary S. Stein, Abdul Rauf Shakoori.
  Journal of Cellular Physiology. June 15, 2016
  Traditional medicinal literature and previous studies have reported the possible role of Cissus quadrangularis (CQ) as an anti‐osteoporotic agent. This study examines the effectiveness of CQ in promoting osteoblast differentiation of the murine pre‐osteoblast cell line, MC3T3‐E1. Ethanolic extract of CQ (CQ‐E) was found to affect growth kinetics of MC3T3‐E1 cells in a dosage‐dependent manner. High concentrations of CQ‐E (more than 10 μg/ml) have particularly adverse effects, while lower concentrations of 0.1 and 1 µg/ml were non‐toxic and did not affect cell viability. Notably, cell proliferation was significantly increased at the lower concentrations of CQ‐E. CQ‐E treatment also augmented osteoblast differentiation, as reflected by a substantial increase in expression of the early osteoblast marker ALP activity, and at later stage, by mineralization of extracellular matrix compared to the control group. These findings suggest dose‐dependent effect of CQ‐E with lower concentrations exhibiting anabolic and osteogenic properties. J. Cell. Physiol. 232: 540–547, 2017. © 2016 Wiley Periodicals, Inc. Ethanolic extract of a herb, Cissus quadrangularis (CQ‐E) has dose dependent effect on the differentiation of murine pre‐osteoblast MC3T3‐E1 cells. Lower concentrations exhibit anabolic and osteogenic properties. We have demonstrated that changes in cellular activity during osteogenesis is accompanied by corresponding changes in expression of osteoblast differentiation specific marker genes. CQ‐E has the potential to be used for healing fractures and to prevent and treat osteoporosis.

  June 15, 2016   doi: 10.1002/jcp.25449   open full text
• Low‐Dose Farnesyltransferase Inhibitor Suppresses HIF‐1α and Snail Expression in Triple‐Negative Breast Cancer MDA‐MB‐231 Cells In Vitro.
  Tomokazu Tanaka, Yuichi Ikegami, Harumasa Nakazawa, Naohide Kuriyama, Miwa Oki, Jun‐ichi Hanai, Vikas P. Sukhatme, Masao Kaneki.
  Journal of Cellular Physiology. June 15, 2016
  The aggressiveness of triple‐negative breast cancer (TNBC), which lacks estrogen receptor, progesterone receptor and epidermal growth factor receptor 2 (HER2), represents a major challenge in breast cancer. Migratory and self‐renewal capabilities are integral components of invasion, metastasis and recurrence of TNBC. Elevated hypoxia‐inducible factor‐1α (HIF‐1α) expression is associated with aggressiveness of cancer. Nonetheless, how HIF‐1α expression is regulated and how HIF‐1α induces aggressive phenotype are not completely understood in TNBC. The cytotoxic effects of farnesyltransferase (FTase) inhibitors (FTIs) have been studied in cancer and leukemia cells. In contrast, the effect of FTIs on HIF‐1α expression has not yet been studied. Here, we show that clinically relevant low‐dose FTI, tipifarnib (300 nM), decreased HIF‐1α expression, migration and tumorsphere formation in human MDA‐MB‐231 TNBC cells under a normoxic condition. In contrast, the low‐dose FTIs did not inhibit cell growth and activity of the Ras pathway in MDA‐MB 231 cells. Tipifarnib‐induced decrease in HIF‐1α expression was associated with amelioration of the Warburg effect, hypermetabolic state, increases in Snail expression and ATP release, and suppressed E‐cadherin expression, major contributors to invasion, metastasis and recurrence of TBNC. These data suggest that FTIs may be capable of ameliorating the aggressive phenotype of TNBC by suppressing the HIF‐1α‐Snail pathway. J. Cell. Physiol. 232: 192–201, 2017. © 2016 Wiley Periodicals, Inc. HIF‐1α and Snail play important roles in metastatic and invasive phenotypes in triple‐negative breast cancer (TNBC). Low‐dose farnesyltransferase inhibitor (FTI) reduced HIF‐1α and Snail expression in TNBC MDA‐MB‐231 cells. In association with the suppression of the HIF‐1α‐Snail pathway, low‐dose FTI decreased migration and sphere formation capabilities in vitro with little, if any, inhibition of the Ras pathway and cell growth in MDA‐MB‐231 cells.

  June 15, 2016   doi: 10.1002/jcp.25411   open full text
• MicroRNA‐590‐5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation.
  M. Vishal, S. Vimalraj, R. Ajeetha, M. Gokulnath, R. Keerthana, Z. He, N.C. Partridge, N. Selvamurugan.
  Journal of Cellular Physiology. June 15, 2016
  Mesenchymal stem cells (MSCs) are multipotent cells and their differentiation into the osteoblastic lineage is strictly controlled by several regulators, including microRNAs (miRNAs). Runx2 is a bone transcription factor required for osteoblast differentiation. Here, we used in silico analysis to identify a number of miRNAs that putatively target Runx2 and its co‐factors to mediate both positive and negative regulation of osteoblast differentiation. Among these miRNAs, miR‐590‐5p was selected and its expression was found to be increased during osteoblast differentiation. When mouse MSCs (mMSCs) were transiently transfected with a miR‐590‐5p mimic, we detected an increase in both calcium deposition and the mRNA expression of osteoblast differentiation marker genes such as alkaline phosphatase (ALP) and type I collagen genes. Smad7 was found to be among the putative target genes of miR‐590‐5p and its mRNA and protein expression decreased after miR‐590‐5p mimic transfection in human osteoblast‐like cells (MG63). Our analysis indicated that Runx2 was not a putative target of miR‐590‐5p. However, Runx2 protein, but not mRNA expression, increased after miR‐590‐5p mimic transfection in MG63 cells. Runx2 protein expression was increased with knockdown of Smad7 expression by Smad7 siRNA in these cells. We further identified that the 3′‐untranslated region of Smad7 was directly targeted by miR‐590‐5p; this was done using the luciferase reporter gene system. It is known that Smad7 inhibits osteoblast differentiation via Smurf2‐mediated Runx2 degradation. Hence, based on our results, we suggest that miR‐590‐5p promotes osteoblast differentiation by indirectly protecting and stabilizing the Runx2 protein by targeting Smad7 gene expression. J. Cell. Physiol. 232: 371–380, 2017. © 2016 Wiley Periodicals, Inc. MiR‐590‐5p promotes osteoblast differentiation by directly targeting Smad7 and indirectly protecting and stabilizing the Runx2 protein from Smurf2‐mediated degradation.

  June 15, 2016   doi: 10.1002/jcp.25434   open full text
• DC‐STAMP: A Key Regulator in Osteoclast Differentiation.
  Ya‐Hui Chiu, Christopher T. Ritchlin.
  Journal of Cellular Physiology. June 14, 2016
  Osteoimmunology research is a new emerging research field that investigates the links between the bone and immune responses. Results from osteoimmunology studies suggest that bone is not only an essential component of the musculoskeletal system, but is also actively involved in immune regulation. Many important factors involved in immune regulation also participate in bone homeostasis. Bone homeostasis is achieved by a coordinated action between bone‐synthesizing osteoblasts and bone‐degrading osteoclasts. An imbalanced action between osteoblasts and osteoclasts often results in pathological bone diseases: osteoporosis is caused by an excessive osteoclast activity, whereas osteopetrosis results from an increased osteoblast activity. This review focuses on dendritic cell‐specific transmembrane protein (DC‐STAMP), an important protein currently considered as a master regulator of osteoclastogenesis. Of clinical relevance, the frequency of circulating DC‐STAMP+ cells is elevated during the pathogenesis of psoriatic diseases. Intriguingly, recent results suggest that DC‐STAMP also plays an imperative role in bone homeostasis by regulating the differentiation of both osteoclasts and osteoblasts. This article summarizes our current knowledge on DC‐STAMP by focusing on its interacting proteins, its regulation on osteoclastogenesis‐related genes, its possible involvement in immunoreceptor tyrosine‐based inhibitory motif (ITIM)‐mediated signaling cascade, and its potential of developing therapeutics for clinical applications. J. Cell. Physiol. 231: 2402–2407, 2016. © 2016 Wiley Periodicals, Inc.

  June 14, 2016   doi: 10.1002/jcp.25389   open full text
• Combination of Rapamycin and Resveratrol for Treatment of Bladder Cancer.
  Anya Alayev, Rachel S. Salamon, Naomi S. Schwartz, Adi Y. Berman, Sara L. Wiener, Marina K. Holz.
  Journal of Cellular Physiology. June 10, 2016
  Loss of TSC1 function, a crucial negative regulator of mTOR signaling, is a common alteration in bladder cancer. Mutations in other members of the PI3K pathway, leading to mTOR activation, are also found in bladder cancer. This provides rationale for targeting mTOR for treatment of bladder cancer characterized by TSC1 mutations and/or mTOR activation. In this study, we asked whether combination treatment with rapamycin and resveratrol could be effective in concurrently inhibiting mTOR and PI3K signaling and inducing cell death in bladder cancer cells. In combination with rapamycin, resveratrol was able to block rapamycin‐induced Akt activation, while maintaining mTOR pathway inhibition. In addition, combination treatment with rapamycin and resveratrol induced cell death specifically in TSC1−/− MEF cells, and not in wild‐type MEFs. Similarly, resveratrol alone or in combination with rapamycin induced cell death in human bladder cancer cell lines. These data indicate that administration of resveratrol together with rapamycin may be a promising therapeutic option for treatment of bladder cancer. J. Cell. Physiol. 232: 436–446, 2017. © 2016 Wiley Periodicals, Inc. Loss of TSC1 function, a crucial negative regulator of mTOR signaling, is a common alteration in bladder cancer, providing rationale for targeting mTOR for treatment of bladder cancer. In this study, we showed that combination treatment with rapamycin and resveratrol is effective in concurrently inhibiting mTOR and PI3K signaling, inducing cell death as well as inhibiting migration and colony formation in bladder cancer cells. These data indicate that administration of resveratrol together with rapamycin may be a promising therapeutic option for treatment of bladder cancer.

  June 10, 2016   doi: 10.1002/jcp.25443   open full text
• Non‐Metabolic Role of PKM2 in Regulation of the HIV‐1 LTR.
  Satarupa Sen, Satish L. Deshmane, Rafal Kaminski, Shohreh Amini, Prasun K. Datta.
  Journal of Cellular Physiology. June 10, 2016
  Identification of cellular proteins, in addition to already known transcription factors such as NF‐κB, Sp1, C‐EBPβ, NFAT, ATF/CREB, and LEF‐1, which interact with the HIV‐1 LTR, is critical in understanding the mechanism of HIV‐1 replication in monocytes/macrophages. Our studies demonstrate upregulation of pyruvate kinase isoform M2 (PKM2) expression during HIV‐1SF162 infection of monocyte/macrophages and reactivation of HIV‐1 in U1 cells, a macrophage model of latency. We observed that HIV‐1SF162 infection of monocyte/macrophages and reactivation of HIV‐1 in U1 cells by PMA resulted in increased levels of nuclear PKM2 compared to PMA‐induced U937 cells. Furthermore, there was a significant increase in the nuclear dimeric form of PKM2 in the PMA‐induced U1 cells in comparison to PMA‐induced U937 cells. We focused on understanding the potential role of PKM2 in HIV‐1 LTR transactivation. Chromatin immunoprecipitation (ChIP) analysis in PMA‐activated U1 and TZM‐bl cells demonstrated the interaction of PKM2 with the HIV‐1 LTR. Our studies show that overexpression of PKM2 results in transactivation of HIV‐1 LTR‐luciferase reporter in U937, U‐87 MG, and TZM‐bl cells. Using various truncated constructs of the HIV‐1 LTR, we mapped the region spanning −120 bp to −80 bp to be essential for PKM2‐mediated transactivation. This region contains the NF‐κB binding site and deletion of this site attenuated PKM2‐mediated activation of HIV‐1 LTR. Immunoprecipitation experiments using U1 cell lysates demonstrated a physical interaction between PKM2 and the p65 subunit of NF‐κB. These observations demonstrate for the first time that PKM2 is a transcriptional co‐activator of HIV‐1 LTR. J. Cell. Physiol. 232: 517–525, 2017. © 2016 Wiley Periodicals, Inc. PKM2 is a transcriptional co‐activator of HIV‐1 LTR.

  June 10, 2016   doi: 10.1002/jcp.25445   open full text
• Effects of Sex and Notch Signaling on the Osteocyte Cell Pool.
  Ernesto Canalis, Lauren Schilling, Stefano Zanotti.
  Journal of Cellular Physiology. June 07, 2016
  Osteocytes play a fundamental role in mechanotransduction and skeletal remodeling. Sex is a determinant of skeletal structure, and female C57BL/6J mice have increased osteoblast number in cancellous bone when compared to male mice. Activation of Notch in the skeleton causes profound cell‐context dependent changes in skeletal physiology. To determine the impact of sex and of Notch signaling on the osteocyte cell pool, we analyzed cancellous and cortical bone of 1–6‐month‐old C57BL/6J or 129SvJ/C57BL/6J mice and determined the osteocyte number/area. There was an age‐dependent decline in osteocyte number in cancellous bone of male but not female mice, so that 6‐month‐old female mice had a greater number of osteocytes than male littermates. Although differences between male and female mice were modest, female mice had ∼10–15% greater number of osteocytes/area. RNA sequence analysis of osteocyte‐rich preparations did not reveal differences between sexes in the expression of genes known to influence bone homeostasis. Neither the activation of Notch1 nor the concomitant inactivation of Notch1 and Notch2 in Osterix (Sp7) or Dentin matrix protein 1 (Dmp1) expressing cells had a pronounced and consistent effect on cancellous or cortical bone osteocyte number in either sex. Moreover, inactivation of Notch1 and Notch2 in Dmp1 expressing cells did not influence the bone loss in a muscle immobilization model of skeletal unloading. In conclusion, cancellous bone osteocytes decline with age in male mice, cortical osteocytes are influenced by sex in younger mice, but osteocyte cell density is not affected substantially by Notch signaling. J. Cell. Physiol. 232: 363–370, 2017. © 2016 Wiley Periodicals, Inc.

  June 07, 2016   doi: 10.1002/jcp.25433   open full text
• Early Treatment With Enalapril and Later Renal Injury in Programmed Obese Adult Rats.
  Hyung Eun Yim, Kee Hwan Yoo, In Sun Bae, Young Sook Hong.
  Journal of Cellular Physiology. June 06, 2016
  Obesity‐related kidney disease should be prevented or retarded. We aimed to investigate whether early treatment with enalapril ameliorates later renal injury induced by early postnatal overnutrition. Three or ten male pups per mother were assigned to either the Obese or Lean group during the first 21 days of life. These pups were treated with enalapril (Obese enalapril, OE; Lean enalapril, LE) or vehicle (Obese control, OC; Lean control, LC) for 15–28 days. Body weight, blood pressure (BP), and renal alterations were determined at 3 months. Enalapril decreased body weight only in the Lean group at 3 months (P < 0.05). Systemic BP levels were higher in the LE, OC, and OE groups than in the LC group at 3 months (P < 0.05). Fewer glomeruli per section area were found in the LE, OC, and OE groups than in the LC group and in the OE group than in the OC group (P < 0.05). The LE and OE groups had higher index scores of glomerulosclerosis and tubulointerstitial fibrosis than the controls (P < 0.05). LE pups showed increased intrarenal angiotensin II receptor type (AT)2 and matrix metalloproteinase (MMP)‐9 and decreased renin and tissue inhibitor of MMP (TIMP)‐1 expression than the LC rats (P < 0.05). OE pups showed increased intrarenal AT2 and decreased AT1 and TIMP‐1 expression than the OC rats (P < 0.05). In conclusion, early treatment with enalapril can induce detrimental renal effects in later life and may not be renoprotective in programmed obese adult rats. J. Cell. Physiol. 232: 447–455, 2017. © 2016 Wiley Periodicals, Inc. Early treatment with enalapril in neonatally overfed rats resulted in decreased glomerular number, increased glomerulosclerosis and tubulointerstitial fibrosis, and dysregulated intrarenal AT1, AT2, and TIMP‐1 expressions later in life. Compared to the Lean control rats, neonatally overfed adult rats were overweight and had hypertension and glomerulosclerosis with a fewer number of glomeruli, regardless of enalapril administration. Moreover, enalapril treatment in Lean rats after the nephrogenic period led to systemic hypertension and progressive renal injury with a low nephron number later in adulthood.

  June 06, 2016   doi: 10.1002/jcp.25444   open full text
• AR‐12 Inhibits Multiple Chaperones Concomitant With Stimulating Autophagosome Formation Collectively Preventing Virus Replication.
  Laurence Booth, Jane L. Roberts, Heath Ecroyd, Sarah R. Tritsch, Sina Bavari, St. Patrick Reid, Stefan Proniuk, Alexander Zukiwski, Abraham Jacob, Claudia S. Sepúlveda, Federico Giovannoni, Cybele C. García, Elsa Damonte, Javier González‐Gallego, María J. Tuñón, Paul Dent.
  Journal of Cellular Physiology. June 06, 2016
  We have recently demonstrated that AR‐12 (OSU‐03012) reduces the function and ATPase activities of multiple HSP90 and HSP70 family chaperones. Combined knock down of chaperones or AR‐12 treatment acted to reduce the expression of virus receptors and essential glucosidase proteins. Combined knock down of chaperones or AR‐12 treatment inactivated mTOR and elevated ATG13 S318 phosphorylation concomitant with inducing an endoplasmic reticulum stress response that in an eIF2α—dependent fashion increased Beclin1 and LC3 expression and autophagosome formation. Over‐expression of chaperones prevented the reduction in receptor/glucosidase expression, mTOR inactivation, the ER stress response, and autophagosome formation. AR‐12 reduced the reproduction of viruses including Mumps, Influenza, Measles, Junín, Rubella, HIV (wild type and protease resistant), and Ebola, an effect replicated by knock down of multiple chaperone proteins. AR‐12—stimulated the co‐localization of Influenza, EBV and HIV virus proteins with LC3 in autophagosomes and reduced viral protein association with the chaperones HSP90, HSP70, and GRP78. Knock down of Beclin1 suppressed drug‐induced autophagosome formation and reduced the anti‐viral protection afforded by AR‐12. In an animal model of hemorrhagic fever virus, a transient exposure of animals to low doses of AR‐12 doubled animal survival from ∼30% to ∼60% and suppressed liver damage as measured by ATL, GGT and LDH release. Thus through inhibition of chaperone protein functions; reducing the production, stability and processing of viral proteins; and stimulating autophagosome formation/viral protein degradation, AR‐12 acts as a broad‐specificity anti‐viral drug in vitro and in vivo. We argue future patient studies with AR‐12 are warranted. J. Cell. Physiol. 231: 2286–2302, 2016. © 2016 Wiley Periodicals, Inc. OSU‐03012 inhibits HSP90 and HSP70 family chaperones which causes enhanced basal levels of endoplasmic reticulum stress and autophagic flux. These biological events collectively result in the destabilization of viral receptors and viral proteins resulting in their degradation. The roles of different chaperones in the replicative potential of viruses as diverse as Measles, HIV, Junin and Ebola has been defined; OSU‐03012 prevents the replication of multiple protease inhibitor resistant strains of HIV. In a rabbit hemorrhagic fever model system, transient exposure to OSU‐03012 reduced animal death and liver damage by over 50%, arguing that the drug has significant anti‐viral activity in vivo.

  June 06, 2016   doi: 10.1002/jcp.25431   open full text
• Milk‐Derived Nanoparticle Fraction Promotes the Formation of Small Osteoclasts But Reduces Bone Resorption.
  Marina C. Oliveira, Irene Di Ceglie, Onno J. Arntz, Wim B. van den Berg, Frank H.J. van den Hoogen, Adaliene V.M. Ferreira, Peter L.E.M. van Lent, Fons A.J. van de Loo.
  Journal of Cellular Physiology. June 02, 2016
  The general consensus is that milk promotes bone growth and density because is a source of calcium and contains components that enhance intestinal calcium uptake or directly affect bone metabolism. In this study, we investigated the effect of bovine‐derived milk 100,000 g pellet (P100), which contains nanoparticles (<220 nm) including extracellular vesicles, on osteoclast differentiation and bone resorption. Bone marrow‐derived osteoclast precursor cells were differentiated into osteoclasts by M‐CSF and RANKL (control) and in the presence of milk P100. Milk P100 treatment until day 4 increased the number of TRAP‐positive mononuclear cells and small (≤5 nuclei) osteoclasts. The number of large (≥6 nuclei) osteoclasts remained the same. These alterations were associated with increased expression of TRAP, NFATc1, and c‐Fos. Cells seeded in a calcium‐phosphate coated plate or bone slices showed reduced resorption area when exposed to milk P100 during the differentiation phase and even after osteoclast formation. Interestingly, milk P100 treatment enhanced Cathepsin K expression but reduced Carbonic Anhydrase 2 gene expression. Moreover, intracellular acid production was also decreased by milk P100 treatment. Oral delivery of milk P100 to female DBA1/J mice for 7 weeks did not alter bone area; however, increased osteoclast number and area in tibia without changes in serum RANKL and CTX‐I levels. We showed for the first time the effect of milk P100 on osteoclast differentiation both in vitro and in vivo and found that milk P100 increased the formation of small osteoclasts but this does not lead to more bone resorption probably due to reduced acid secretion. J. Cell. Physiol. 232: 225–233, 2017. © 2016 Wiley Periodicals, Inc. Milk nanoparticle fraction treatment increases osteoclast differentiation into small cells. Osteoclast activity is impaired with milk nanoparticle fraction treatment. Milk nanocomponents influence bone remodeling.

  June 02, 2016   doi: 10.1002/jcp.25414   open full text
• How Chimeric Antigen Receptor Design Affects Adoptive T Cell Therapy.
  Albert T. Gacerez, Benjamine Arellano, Charles L. Sentman.
  Journal of Cellular Physiology. June 02, 2016
  Chimeric antigen receptor (CAR) T cells have been developed to treat tumors and have shown great success against B cell malignancies. Exploiting modular designs and swappable domains, CARs can target an array of cell surface antigens and, upon receptor‐ligand interactions, direct signaling cascades, thereby driving T cell effector functions. CARs have been designed using receptors, ligands, or scFv binding domains. Different regions of a CAR have each been found to play a role in determining the overall efficacy of CAR T cells. Therefore, this review provides an overview of CAR construction and common designs. Each CAR region is discussed in the context of its importance to a CAR's function. Additionally, the review explores how various engineering strategies have been applied to CAR T cells in order to regulate CAR T cell function and activity. J. Cell. Physiol. 231: 2590–2598, 2016. © 2016 Wiley Periodicals, Inc. Chimeric antigen receptor (CAR) T cells are able to eliminate late stage hematological cancers and have a great potential to treat many types of cancer. This review summarizes how the design of CARs alters their function and T cell responses.

  June 02, 2016   doi: 10.1002/jcp.25419   open full text
• Repression of Primitive Erythroid Program Is Critical for the Initiation of Multi‐Lineage Hematopoiesis in Mouse Development.
  Toshiyuki Yamane, Chie Ito, Aya Washino, Kana Isono, Hidetoshi Yamazaki.
  Journal of Cellular Physiology. June 02, 2016
  Formation of the hematopoietic cells occurs in multiple steps. The first hematopoietic cells observed during ontogeny are primitive erythrocytes, which are produced in the early yolk sac within a limited temporal window. Multi‐lineage hematopoiesis, which supplies almost the entire repertoire of blood cell lineages, lags behind primitive erythropoiesis in the tissue. However, molecular mechanisms regulating sequential generation of primitive erythrocytes and multipotent hematopoietic progenitors in the yolk sac are largely unknown. In this study, the transcription factors involved in the development of hematopoietic cells were examined in purified progenitor cell populations from pluripotent stem cell cultures and from the yolk sac of developing embryos. We found that the earliest committed hematopoietic progenitors highly expressed Gata1, Scl/tal1, and Klf1 genes. Expression of these transcription factors, which is known to form a core erythroid transcriptional network, explained the prompt generation of primitive erythrocytes from these earliest progenitors. Importantly, the multipotent hematopoietic cells, which lack the differentiation potential into primitive erythroid cells, down‐regulated these genes during a transition from the earliest committed progenitors. In addition, we showed that Pu.1 is involved in the multipotent cell differentiation through the suppression of erythroid transcription program. We propose that these molecular mechanisms governed by transcription factors form sequential waves of primitive erythropoiesis and multi‐lineage hematopoiesis in the early yolk sac of developing embryos. J. Cell. Physiol. 232: 323–330, 2017. © 2016 Wiley Periodicals, Inc. The first blood cell progenitors highly expressed red blood cell‐specific transcription factors. This expression of transcription factors was considered to enable the immediate production of primitive red blood cells. Importantly, the multipotent blood cell progenitors derived from the earliest progenitors shut‐off the expression of red blood cell‐specific transcription factors, which was partly mediated through the upregulation of Pu.1 expression.

  June 02, 2016   doi: 10.1002/jcp.25422   open full text
• Flavopiridol: An Old Drug With New Perspectives? Implication for Development of New Drugs.
  Annamaria Cimini, Michele d'Angelo, Elisabetta Benedetti, Barbara D'Angelo, Giulio Laurenti, Andrea Antonosante, Loredana Cristiano, Antonella Di Mambro, Marcella Barbarino, Vanessa Castelli, Benedetta Cinque, Maria Grazia Cifone, Rodolfo Ippoliti, Francesca Pentimalli, Antonio Giordano.
  Journal of Cellular Physiology. June 02, 2016
  Glioblastoma, the most common brain tumor, is characterized by high proliferation rate, invasion, angiogenesis, and chemo‐ and radio‐resistance. One of most remarkable feature of glioblastoma is the switch toward a glycolytic energetic metabolism that leads to high glucose uptake and consumption and a strong production of lactate. Activation of several oncogene pathways like Akt, c‐myc, and ras induces glycolysis and angiogenesis and acts to assure glycolysis prosecution, tumor proliferation, and resistance to therapy. Therefore, the high glycolytic flux depends on the overexpression of glycolysis‐related genes resulting in an overproduction of pyruvate and lactate. Metabolism of glioblastoma thus represents a key issue for cancer research. Flavopiridol is a synthetic flavonoid that inhibits a wide range of Cyclin‐dependent kinase, that has been demonstrate to inactivate glycogen phosphorylase, decreasing glucose availability for glycolysis. In this work the study of glucose metabolism upon flavopiridol treatment in the two different glioblastoma cell lines. The results obtained point towards an effect of flavopiridol in glycolytic cells, thus suggesting a possible new use of this compound or flavopiridol‐derived formulations in combination with anti‐proliferative agents in glioblastoma patients. J. Cell. Physiol. 232: 312–322, 2017. © 2016 Wiley Periodicals, Inc. FLAP treatment affects glucose utilization pathways in two glioblastoma cell lines, bearing different PTEN status. The treatment strongly decreases the glycolytic enzymes with concomitant increase of cell death.

  June 02, 2016   doi: 10.1002/jcp.25421   open full text
• Sex Differences in Estrogen Receptor α and β Levels and Activation Status in LPS‐Stimulated Human Macrophages.
  Ilaria Campesi, Maria Marino, Andrea Montella, Sara Pais, Flavia Franconi.
  Journal of Cellular Physiology. June 02, 2016
  Immune function, inflammation, and atherosclerosis display sex differences and are influenced by 17β‐estradiol through estrogen receptors subtypes ERα and ERβ. Male tissues express active ERs, but their possible involvement in inflammation in males has never been assessed. Macrophages express both ERα and ERβ and offer the opportunity to evaluate the role of ER levels and activation in inflammation. We assessed the ability of lipopolysaccharide (LPS) to modulate, in a sex‐specific way, the expression and the activation status of ERα and ERβ in blood monocytes‐derived macrophages (MDMs) from men and women. MDMs were incubated with 100 ng/ml LPS for 24 h and used to evaluate ERα, ERβ, P‐ERα, p38, and P‐p38 expression by Western Blotting. In basal conditions, ERα and ERβ were significantly higher in female MDMs than in male MDMs. LPS up‐regulated ERα and ERα phosphorylation in both sexes, with a significantly higher effect observed in male MDMs, and down‐regulated ERβ level only in female MDMs. p38 and P‐p38 proteins, indicative of ERβ activity, did not show sex differences both in basal conditions and after LPS treatment. Finally, ERα/ERβ and P‐ERα/ERα ratios were significantly higher in male MDMs than in female ones. Our data indicate, for the first time, that LPS affects ERα but not ERβ activation status. We identify a significant role of ERα in LPS‐mediated inflammatory responses in MDMs, which represents an initial step in understanding the influence of sex in the relationship between LPS and ERα. J. Cell. Physiol. 232: 340–345, 2017. © 2016 Wiley Periodicals, Inc.

  June 02, 2016   doi: 10.1002/jcp.25425   open full text
• Chrysophanol Induces Apoptosis of Choriocarcinoma Through Regulation of ROS and the AKT and ERK1/2 Pathways.
  Whasun Lim, Changwon Yang, Fuller W. Bazer, Gwonhwa Song.
  Journal of Cellular Physiology. June 02, 2016
  Chrysophanol is an anthraquinone compound, mainly isolated from rhubarb, with anti‐cancer effects on some types of cancer cells. However, effects of chrysophanol on human choriocarcinoma cells are not known. Therefore, the objective of this study was to determine effects of chrysophanol on choriocarcinoma cells (JAR and JEG‐3) and identify signal transduction cascades activated by chrysophanol. Results of present study, showed that chrysophanol decreased cell viability and induced apoptosis of JEG‐3, but not JAR cells, in a dose‐dependent manner. Chrysophanol also increased oxidative stress in JEG‐3 cells by inducing ROS generation followed by mitochondrial dysfunction including depolarization of mitochondrial inner membrane potential. Western blot analysis revealed that ERK1/2, P90RSK, AKT, and P70S6K were increased significantly in JEG‐3 cells by chrysophanol. Next, we investigated chrysophanol‐mediated effects on proliferation of JEG‐3 cells using pharmacological inhibitors of PI3K/AKT (LY294002) and ERK1/2 (U0126). Inhibition of AKT and ERK1/2 prevented chrysophanol‐induced stimulation of proliferation of JEG‐3 cells. In addition, the phosphorylation of AKT and ERK1/2 was suppressed by LY294002 and U0126 in JEG‐3 cells treated with chrysophanol, whereas, the AKT protein was activated by pre‐treatment of JEG‐3 cells with U0126. Furthermore, we compared therapeutic effects of chrysophanol with cisplatin and paclitaxel which are conventional salvage regimens for choriocarcinoma. Our results verified that chrysophanol has synergistic effects with traditional therapy to increase apoptosis of JEG‐3 cells. Collectively, these results indicate that chrysophanol is a potential effective chometherapeutic agent for treatment of choriocarcinoma therapy, and minimizing side effects of conventional treatment regimens. J. Cell. Physiol. 232: 331–339, 2017. © 2016 Wiley Periodicals, Inc. Results of the present study indicated that chrysophanol has anti‐cancer effects by reducing cell viability and inducing apoptosis by increasing ROS production and decreasing mitochondrial membrane potential through the ERK1/2 and AKT signaling cascades. Moreover, chrysophanol is a potential effective chometherapeutic agent for treatment of choriocarcinoma therapy when combined with cisplatin and paclitaxel and minimizing side effects of conventional treatment regimens.

  June 02, 2016   doi: 10.1002/jcp.25423   open full text
• Ginkgolic Acid Inhibits Invasion and Migration and TGF‐β‐Induced EMT of Lung Cancer Cells Through PI3K/Akt/mTOR Inactivation.
  Seung Ho Baek, Jeong‐Hyeon Ko, Jong Hyun Lee, Chulwon Kim, Hanwool Lee, Dongwoo Nam, Junhee Lee, Seok‐Geun Lee, Woong Mo Yang, Jae‐Young Um, Gautam Sethi, Kwang Seok Ahn.
  Journal of Cellular Physiology. June 02, 2016
  Epithelial‐to‐mesenchymal transition (EMT) is a critical cellular phenomenon regulating tumor metastases. In the present study, we investigated whether ginkgolic acid can affect EMT in lung cancer cells and the related underlying mechanism(s) of its actions. We found that ginkgolic acid C15:1 (GA C15:1) inhibited cell proliferation, invasion, and migration in both A549 and H1299 lung cancer cells. GA C15:1 also suppressed the expression of EMT related genes (Fibronectin, Vimentin, N‐cadherin, MMP‐9, MMP‐2, Twist and Snail) and suppressed TGF‐β‐induced EMT as assessed by reduced expression of mesenchymal markers (Fibronectin, Vimentin, N‐cadherin), MMP‐9, MMP‐2, Twist and Snail. However, GA C15:1 did not affect the expression of various epithelial marker proteins (Occludin and E‐cadherin) in both A549 and H1299 cells. TGF‐β‐induced morphologic changes from epithelial to mesenchymal cells and induction of invasion and migration were reversed by GA C15:1. Finally, GA C15:1 not only abrogated basal PI3K/Akt/mTOR signaling cascade, but also reduced TGF‐β‐induced phosphorylation of PI3K/Akt/mTOR pathway in lung cancer cells. Overall, these findings suggest that GA C15:1 suppresses lung cancer invasion and migration through the inhibition of PI3K/Akt/mTOR signaling pathway and provide a source of potential therapeutic compounds to control the metastatic dissemination of tumor cells. J. Cell. Physiol. 232: 346–354, 2017. © 2016 Wiley Periodicals, Inc. GA C15:1 also suppressed the expression of EMT related genes. GA C15:1 suppressed TGF‐beta‐induced mesenchymal markers, MMP‐9, MMP‐2, Twist and Snail. GA C15:1 suppresses metastatic lung cancer through inhibition of PI3K/Akt/mTOR activation.

  June 02, 2016   doi: 10.1002/jcp.25426   open full text
• Anti‐Diabetic Agent Sodium Tungstate Induces the Secretion of Pro‐ and Anti‐Inflammatory Cytokines by Human Kidney Cells.
  Romina Bertinat, Francisco Westermeier, Pamela Silva, Jie Shi, Francisco Nualart, Xuhang Li, Alejandro J. Yáñez.
  Journal of Cellular Physiology. June 02, 2016
  Diabetic kidney disease (DKD) is the major cause of end stage renal disease. Sodium tungstate (NaW) exerts anti‐diabetic and immunomodulatory activities in diabetic animal models. Here, we used primary cultures of renal proximal tubule epithelial cells derived from type‐2‐diabetic (D‐RPTEC) and non‐diabetic (N‐RPTEC) subjects as in vitro models to study the effects of NaW on cytokine secretion, as these factors participate in intercellular regulation of inflammation, cell growth and death, differentiation, angiogenesis, development, and repair, all processes that are dysregulated during DKD. In basal conditions, D‐RPTEC cells secreted higher levels of prototypical pro‐inflammatory IL‐6, IL‐8, and MCP‐1 than N‐RPTEC cells, in agreement with their diabetic phenotype. Unexpectedly, NaW further induced IL‐6, IL‐8, and MCP‐1 secretion in both N‐ and D‐RPTEC, together with lower levels of IL‐1 RA, IL‐4, IL‐10, and GM‐CSF, suggesting that it may contribute to the extent of renal damage/repair during DKD. Besides, NaW induced the accumulation of IκBα, the main inhibitor protein of one major pathway involved in cytokine production, suggesting further anti‐inflammatory effect in the long‐term. A better understanding of the mechanisms involved in the interplay between the anti‐diabetic and immunomodulatory properties of NaW will facilitate future studies about its clinical relevance. J. Cell. Physiol. 232: 355–362, 2017. © 2016 Wiley Periodicals, Inc. Sodium tungstate (NaW) exerts anti‐diabetic activities. We further show that NaW induces IL‐6, IL‐8, and MCP‐1 secretion, together with lower levels of IL1‐RA, IL‐4, IL‐10, GM‐CSF in renal proximal tubule epithelial cells from diabetic and non‐diabetic subjects, suggesting that it may contribute to the extent of renal damage/repair of diabetic kidney.

  June 02, 2016   doi: 10.1002/jcp.25429   open full text
• Neoadjuvant Sequential Docetaxel Followed by High‐Dose Epirubicin in Combination With Cyclophosphamide Administered Concurrently With Trastuzumab. The DECT Trial.
  Laura Pizzuti, Maddalena Barba, Diana Giannarelli, Domenico Sergi, Claudio Botti, Paolo Marchetti, Michele Anzà, Marcello Maugeri‐Saccà, Clara Natoli, Simona Di Filippo, Teresa Catenaro, Federica Tomao, Antonella Amodio, Silvia Carpano, Letizia Perracchio, Marcella Mottolese, Luigi Di Lauro, Giuseppe Sanguineti, Anna Di Benedetto, Antonio Giordano, Patrizia Vici.
  Journal of Cellular Physiology. June 02, 2016
  To report the results of the DECT trial, a phase II study of locally advanced or operable HER2‐positive breast cancer (BC) treated with taxanes and concurrent anthracyclines and trastuzumab. Eligible patients (stage IIA‐IIIB HER2‐positive BC, 18–75 years, normal organ functions, ECOG ≤1, and left ventricular ejection fraction (LVEF) ≥55%) received four cycles of neoadjuvant docetaxel, 100 mg/m2 intravenously, plus trastuzumab 6 mg/kg (loading dose 8 mg/kg) every 3 weeks, followed by four 3‐weekly cycles of epirubicin 120 mg/m2 and cyclophosphamide, 600 mg/m2, plus trastuzumab. Primary objective was pathologic complete response (pCR) rate, defined as ypT0/is ypN0 at definitive surgery. We enrolled 45 consecutive patients. All but six patients (13.3%) completed chemotherapy and all underwent surgery. pCR was observed in 28 patients (62.2%) overall and in 6 (66.7%) from the inflammatory subgroup. The classification and regression tree analysis showed a 100% pCR rate in patients with BMI ≥25 and with hormone negative disease. The median follow up was 46 months (8–78). Four‐year recurrence‐free survival was 74.7% (95%CI, 58.2–91.2). Seven patients (15.6%) recurred and one died. Treatment was well tolerated, with limiting toxicity being neutropenia. No clinical cardiotoxicity was observed. Six patients (13.4%) showed a transient LVEF decrease (<10%). In one patient we observed a ≥10% asymptomatic LVEF decrease persisting after surgery. Notwithstanding their limited applicability due to the current guidelines, our findings support the efficacy of the regimen of interest in the neoadjuvant setting along with a fairly acceptable toxicity profile, including cardiotoxicity. Results on BMI may invite further assessment in future studies. J. Cell. Physiol. 231: 2541–2547, 2016. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. The DECT trial supports the efficacy of the combined use of epirubicin and trastuzumab in the neoadjuvant setting along with a fairly acceptable toxicity profile, including cardiotoxicity.

  June 02, 2016   doi: 10.1002/jcp.25432   open full text
• Generation of Neural Crest‐Like Cells From Human Periodontal Ligament Cell‐Derived Induced Pluripotent Stem Cells.
  Atsushi Tomokiyo, Kim Hynes, Jia Ng, Danijela Menicanin, Esther Camp, Agnes Arthur, Stan Gronthos, Peter Mark Bartold.
  Journal of Cellular Physiology. June 02, 2016
  Neural crest cells (NCC) hold great promise for tissue engineering, however the inability to easily obtain large numbers of NCC is a major factor limiting their use in studies of regenerative medicine. Induced pluripotent stem cells (iPSC) are emerging as a novel candidate that could provide an unlimited source of NCC. In the present study, we examined the potential of neural crest tissue‐derived periodontal ligament (PDL) iPSC to differentiate into neural crest‐like cells (NCLC) relative to iPSC generated from a non‐neural crest derived tissue, foreskin fibroblasts (FF). We detected high HNK1 expression during the differentiation of PDL and FF iPSC into NCLC as a marker for enriching for a population of cells with NCC characteristics. We isolated PDL iPSC‐ and FF iPSC‐derived NCLC, which highly expressed HNK1. A high proportion of the HNK1‐positive cell populations generated, expressed the MSC markers, whilst very few cells expressed the pluripotency markers or the hematopoietic markers. The PDL and FF HNK1‐positive populations gave rise to smooth muscle, neural, glial, osteoblastic and adipocytic like cells and exhibited higher expression of smooth muscle, neural, and glial cell‐associated markers than the PDL and FF HNK1‐negative populations. Interestingly, the HNK1‐positive cells derived from the PDL‐iPSC exhibited a greater ability to differentiate into smooth muscle, neural, glial cells and adipocytes, than the HNK1‐positive cells derived from the FF‐iPSC. Our work suggests that HNK1‐enriched NCLC from neural crest tissue‐derived iPSC more closely resemble the phenotypic and functional hallmarks of NCC compared to the HNK1‐low population and non‐neural crest iPSC‐derived NCLC. J. Cell. Physiol. 232: 402–416, 2017. © 2016 Wiley Periodicals, Inc. The aim of this study was to generate the cells that closely resemble the phenotypic and functional hallmarks of neural crest cells using iPSC. We demonstrated that iPSC derived from neural crest lineage cells a greater capacity to differentiate into neural crest‐like cells than iPSC derived from non‐neural crest lineage cells. We suggested that cell sorting using the HNK1 antibody is an efficient method to enrich a population of cells with neural crest cell characteristics.

  June 02, 2016   doi: 10.1002/jcp.25437   open full text
• Epithelial‐to‐Mesenchymal Transition in Paclitaxel‐Resistant Ovarian Cancer Cells Is Downregulated by Luteolin.
  Vermont P Dia, Philipus Pangloli.
  Journal of Cellular Physiology. May 27, 2016
  Ovarian cancer (OVCA) is the deadliest of all gynecological cancers which is attributed to late presentation, persistence, and development of chemoresistance. The objectives were to evaluate the association between OVCA paclitaxel‐resistance and epithelial‐to‐mesenchymal transition (EMT) and to determine the capability of luteolin to chemosensitize OVCA cells. X10 and X22 cells were 11.8–25.3‐fold and 7.8–8.6‐fold resistant to paclitaxel than 1AP cells. X10 and X22 cells exhibited a mesenchymal phenotype, while 1AP has an epithelial characteristics. Furthermore, the expression of the epithelial marker E‐cadherin was downregulated, while mesenchymal markers Vimentin and N‐cadherin were upregulated in X10 and X22 cells when compared to 1AP cells. Transcription factors Snail, Slug, and Twist1 were upregulated in X10 cells, while Twist1 was highly expressed in X22 cells. Luteolin treatment caused cytotoxicity being most potent to X10 OVCA cells. Treatment of non‐cytotoxic dose of luteolin at 15.625 μM chemosensitized X10 and X22 OVCA cells to paclitaxel as evidenced by reduced ED50 values from 11.8 to 0.2 μM and 8.6 to 3.6 μM for X10 and X22 cells, respectively. Moreover, luteolin treatment led to a more epithelial phenotype of X10 and X22 cells and modification of EMT markers indicating reversal of EMT. The mechanism involved is through reduction of phosphorylation of FAK and ERK leading to reduced nuclear translocation of p65. Our results highlight the significance of EMT in OVCA resistance to paclitaxel and warrant the investigation of luteolin as a potential therapeutic agent in chemoresistant OVCA. J. Cell. Physiol. 232: 391–401, 2017. © 2016 Wiley Periodicals, Inc. Epithelial‐to‐mesenchymal transition (EMT) is upregulated in paclitaxel‐resistant ovarian cancer cells. Luteolin chemosensitized paclitaxel‐resistant ovarian cancer cells. This chemosensitization may be associated with modification of EMT markers upon luteolin treatment.

  May 27, 2016   doi: 10.1002/jcp.25436   open full text
• Prostaglandin E2‐Dependent Phosphorylation of RAS Inhibition 1 (RIN1) at Ser 291 and 292 Inhibits Transforming Growth Factor‐β‐Induced RAS Activation Pathway in Human Synovial Fibroblasts: Role in Cell Migration.
  Casimiro Gerarduzzi, QingWen He, Beibei Zhai, John Antoniou, John A. Di Battista.
  Journal of Cellular Physiology. May 26, 2016
  Prostaglandin E2 (PGE2)‐stimulated G‐protein–coupled receptor (GPCR) activation inhibits pro‐fibrotic TGFβ‐dependent stimulation of human fibroblast to myofibroblast transition (FMT), though the precise molecular mechanisms are not fully understood. In the present study, we describe the PGE2‐dependent suppression and reversal of TGFβ‐induced events such as α‐sma expression, stress fiber formation, and Ras/Raf/ERK/MAPK pathway‐dependent activation of myofibroblast migration. In order to elucidate post‐ligand‐receptor signaling pathways, we identified a predominant PKA phosphorylation motif profile in human primary fibroblasts after treatment with exogenous PGE2 (EC50 30 nM, Vmax 100 nM), mimicked by the adenyl cyclase activator forskolin (EC50 5 μM, Vmax 10 μM). We used a global phosphoproteomic approach to identify a 2.5‐fold difference in PGE2‐induced phosphorylation of proteins containing the PKA motif. Deducing the signaling pathway of our migration data, we identified Ras inhibitor 1 (RIN1) as a substrate, whereby PGE2 induced its phosphorylation at Ser291 and at Ser292 by a 5.4‐ and 4.8‐fold increase, respectively. In a series of transient and stable over expression studies in HEK293T and HeLa cells using wild‐type (wt) and mutant RIN1 (Ser291/292Ala) or Ras constructs and siRNA knock‐down experiments, we showed that PGE2‐dependent phosphorylation of RIN1 resulted in the abrogation of TGFβ‐induced Ras/Raf signaling activation and subsequent downstream blockade of cellular migration, emphasizing the importance of such phosphosites in PGE2 suppression of wound closure. Overexpression experiments in tandem with pull‐down assays indicated that specific Ser291/292 phosphorylation of RIN1 favored binding to activated Ras. In principal, understanding PGE2‐GPCR activated signaling pathways mitigating TGFβ‐induced fibrosis may lead to more evidence‐based treatments against the disease. J. Cell. Physiol. 232: 202–215, 2017. © 2016 Wiley Periodicals, Inc. Background: PGE2 is an antifibrotic agent with a limited molecular mechanism of action against myofibroblasts. Results: PGE2 induces the phosphorylation of RIN1 at Ser291/292 to inhibit the TGFβ‐activated Ras/Raf pathway of myofibroblast migration. Conclusion: An antifibrotic mechanism of PGE2 blocks myofibroblast migration through RIN1. Significance: Understanding the endogenous mechanisms of antifibrosis could be extrapolated for therapeutic means.

  May 26, 2016   doi: 10.1002/jcp.25412   open full text
• LPS Induces Hyper‐Permeability of Intestinal Epithelial Cells.
  Amir Bein, Alexander Zilbershtein, Michael Golosovsky, Dan Davidov, Betty Schwartz.
  Journal of Cellular Physiology. May 26, 2016
  Necrotizing Enterocolitis (NEC) is a severe inflammatory disorder leading to high morbidity and mortality rates. A growing body of evidence demonstrate the key role of the Toll like receptor 4 (TLR4) in NEC. This membranal receptor recognizes lipopolysaccharides (LPS) from the bacterial wall and triggers an inflammatory response. The aim of the present study was to elucidate the effect of LPS on paracellular permeability known to be severely affected in NEC. IEC‐18 cells were treated with LPS and the effects on morphology, paracellular permeability and their associated gene and protein expressions were measured. Our results show that LPS down regulated the expression of occludin and ZO‐1 mRNAs while up regulating Cdkn1a. In addition LPS caused a significant increase in paracellular permeability and epithelial barrier damage. Finally ZO‐1 protein was found to be spatially disarrayed in the intercellular junctions in response to LPS. We conclude that LPS adversely affected the functionality of the intestinal epithelial barrier suggesting a new mechanism by which bacterial infection may contribute to the development of NEC. J. Cell. Physiol. 232: 381–390, 2017. © 2016 Wiley Periodicals, Inc. This study explores the effect of LPS on paracellular permeability known to be severely affected in NEC. We show that LPS adversely affected the functionality of the intestinal epithelial barrier and in particular led to spatial disarrangement of ZO‐1 protein.

  May 26, 2016   doi: 10.1002/jcp.25435   open full text
• IDO‐Expressing Fibroblasts Protect Islet Beta Cells From Immunological Attack and Reverse Hyperglycemia in Non‐Obese Diabetic Mice.
  Yun Zhang, Reza B. Jalili, Ruhangiz T. Kilani, Sanam Salimi Elizei, Ali Farrokhi, Mohsen Khosravi‐Maharlooei, Garth L. Warnock, Ziliang Ao, Lucy Marzban, Aziz Ghahary.
  Journal of Cellular Physiology. May 08, 2016
  Indoleamine 2,3‐dioxygenase (IDO) induces immunological tolerance in physiological and pathological conditions. Therefore, we used dermal fibroblasts with stable IDO expression as a cell therapy to: (i) Investigate the factors determining the efficacy of this cell therapy for autoimmune diabetes in non‐obese diabetic (NOD) mice; (ii) Scrutinize the potential immunological mechanisms. Newly diabetic NOD mice were randomly injected with either 10 × 106 (10M) or 15 × 106 (15M) IDO‐expressing dermal fibroblasts. Blood glucose levels (BGLs), body weight, plasma kynurenine levels, insulitis severity, islet beta cell function, autoreactive CD8+ T cells, Th17 cells and regulatory T cells (Tregs) were then investigated in these mice. IL‐1β and cleaved caspase‐3 levels were assessed in islets co‐cultured with IDO‐expressing fibroblasts. BGLs in 83% mice treated with 15M IDO‐expressing fibroblasts recovered to normal up to 120 days. However, only 17% mice treated with 10M IDO‐expressing cells were reversed to normoglycemia. A 15M IDO‐expressing fibroblasts significantly reduced infiltrated immune cells in islets and recovered the functionality of remaining islet beta cells in NOD mice. Additionally, they successfully inhibited autoreactive CD8+ T cells and Th17 cells as well as increased Tregs in different organs of NOD mice. Islet beta cells co‐cultured with IDO‐expressing fibroblasts had reduced IL‐1β levels and cell apoptosis. Both cell number and IDO enzymatic activity contributes to the efficiency of IDO cell therapy. Optimized IDO‐expressing fibroblasts successfully reverse the progression of diabetes in NOD mice through induction of Tregs as well as inhibition of beta cell specific autoreactive CD8+ T cells and Th17 cells. J. Cell. Physiol. 231: 1964–1973, 2016. © 2016 Wiley Periodicals, Inc.

  May 08, 2016   doi: 10.1002/jcp.25301   open full text
• SHP‐2 Phosphatase Prevents Colonic Inflammation by Controlling Secretory Cell Differentiation and Maintaining Host‐Microbiota Homeostasis.
  Geneviève Coulombe, Ariane Langlois, Giada De Palma, Marie‐Josée Langlois, Justin L. McCarville, Jessica Gagné‐Sanfaçon, Nathalie Perreault, Gen‐Sheng Feng, Premysl Bercik, François Boudreau, Elena F. Verdu, Nathalie Rivard.
  Journal of Cellular Physiology. May 08, 2016
  Polymorphisms in the PTPN11 gene encoding for the tyrosine phosphatase SHP‐2 were described in patients with ulcerative colitis. We have recently demonstrated that mice with an intestinal epithelial cell‐specific deletion of SHP‐2 (SHP‐2IEC‐KO) develop severe colitis 1 month after birth. However, the mechanisms by which SHP‐2 deletion induces colonic inflammation remain to be elucidated. We generated SHP‐2IEC‐KO mice lacking Myd88 exclusively in the intestinal epithelium. The colonic phenotype was histologically analyzed and cell differentiation was determined by electron microscopy and lysozyme or Alcian blue staining. Microbiota composition was analyzed by 16S sequencing. Results show that innate defense genes including those specific to Paneth cells were strongly up‐regulated in SHP‐2‐deficient colons. Expansion of intermediate cells (common progenitors of the Goblet and Paneth cell lineages) was found in the colon of SHP‐2IEC‐KO mice whereas Goblet cell number was clearly diminished. These alterations in Goblet/intermediate cell ratio were noticed 2 weeks after birth, before the onset of inflammation and were associated with significant alterations in microbiota composition. Indeed, an increase in Enterobacteriaceae and a decrease in Firmicutes were observed in the colon of these mice, indicating that dysbiosis also occurred prior to inflammation. Importantly, loss of epithelial Myd88 expression inhibited colitis development in SHP‐2IEC‐KO mice, rescued Goblet/intermediate cell ratio, and prevented NFκB hyperactivation and inflammation. These data indicate that SHP‐2 is functionally important for the maintenance of appropriate barrier function and host‐microbiota homeostasis in the large intestine. J. Cell. Physiol. 231: 2529–2540, 2016. © 2016 The Authors. Journal of Cellular Physiology published by Wiley Periodicals, Inc. Variations within the gene locus encoding SHP‐2 (PTPN11) have been associated with increased susceptibility to develop ulcerative colitis. In this regard, mice with an intestinal epithelial cell‐specific deletion of SHP‐2 expression rapidly develop colitis. The results indicate that SHP‐2 prevents colonic inflammation by controlling secretory cell differentiation and maintaining host‐ microbiota homeostasis.

  May 08, 2016   doi: 10.1002/jcp.25407   open full text
• Research Needs for Understanding the Biology of Overdiagnosis in Cancer Screening.
  Sudhir Srivastava, Brian J. Reid, Sharmistha Ghosh, Barnett S. Kramer.
  Journal of Cellular Physiology. April 29, 2016
  Many cancers offer an extended window of opportunity for early detection and therapeutic intervention that could lead to a reduction in cause‐specific mortality. The pursuit of early detection in screening settings has resulted in decreased incidence and mortality for some cancers (e.g., colon and cervical cancers), and increased incidence with only modest or no effect on cause‐specific mortality in others (e.g., breast and prostate). Whereas highly sensitive screening technologies are better at detecting a number of suspected “cancers” that are indolent and likely to remain clinically unimportant in the lifetime of a patient, defined as overdiagnosis, they often miss cancers that are aggressive and tend to present clinically between screenings, known as interval cancers. Unrecognized overdiagnosis leads to overtreatment with its attendant (often long‐lasting) side effects, anxiety, and substantial financial harm. Existing methods often cannot differentiate indolent lesions from aggressive ones or understand the dynamics of neoplastic progression. To correctly identify the population that would benefit the most from screening and identify the lesions that would benefit most from treatment, the evolving genomic and molecular profiles of individual cancers during the clinical course of progression or indolence must be investigated, while taking into account an individual's genetic susceptibility, clinical and environmental risk factors, and the tumor microenvironment. Practical challenges lie not only in the lack of access to tissue specimens that are appropriate for the study of natural history, but also in the absence of targeted research strategies. This commentary summarizes the recommendations from a diverse group of scientists with expertise in basic biology, translational research, clinical research, statistics, and epidemiology and public health professionals convened to discuss research directions. J. Cell. Physiol. 231: 1870–1875, 2016. © 2015 Wiley Periodicals, Inc.

  April 29, 2016   doi: 10.1002/jcp.25227   open full text
• Antibodies Against Mimotopes of Simian Virus 40 Large T Antigen, the Oncoprotein, in Serum Samples From Elderly Healthy Subjects.
  Elisa Mazzoni, Giovanni Guerra, Maria Vittoria Casali, Silvia Pietrobon, Ilaria Bononi, Andrea Puozzo, Andrea Tagliapietra, Pier Francesco Nocini, Mauro Tognon, Fernanda Martini.
  Journal of Cellular Physiology. April 29, 2016
  Simian Virus 40 (SV40), a monkey polyomavirus, was administered to human populations by early anti‐poliomylitis vaccines contaminated by this small DNA tumor virus. Data on SV40 infection in humans remain controversial. Elderly subjects represent an interesting cohort to investigate, because they were not immunized with SV40‐contaminated vaccines. Taking advantage of the Italian population, the second oldest worldwide, elderly subjects (n = 237) up to 100 years old were enrolled in this study. Their sera were analyzed, by ELISA tests with synthetic peptides mimicking the viral epitopes, for IgG antibodies reacting with SV40 large Tumor antigen (Tag), the viral oncoprotein. An overall seroprevalence of 22% was revealed in subjects aged 66–100 years, ranging from 19% in individuals 66–74 years old, to 24% in subjects 82–100 years old, with a lower SV40 titer detected in the oldest group. Our data show that: (i) SV40 infection is not frequent in old individuals; (ii) the infection rate increases in elderly with the age; (iii) the antibody titer of SV40 Tag decreases with the age. In conclusion, SV40 infection seems to spread in old subjects independently from SV40‐contaminated vaccines. This study seems to confirm that SV40 is also a human virus. J. Cell. Physiol. 232: 176–181, 2017. © 2016 Wiley Periodicals, Inc. Specific antibodies reacting against Simian Virus 40 large T antigen mimotopes were detected in serum samples from elderly healthy subjects, up to 100 years old, with a prevalence of 22%. Immunological data were obtained with a novel indirect ELISA with synthetic peptides, which mimic the immunogenic epitopes of the viral oncoprotein of SV40. Our data suggests that the exposure to Simian Virus 40 infection is not frequent in old individuals. The immunological results showed that (i) the infection rate increases in elderly with the age; (ii) the proportion of elderly subjects with high Simian Virus 40 large T antigen antibody levels decreases with age; (iii) Simian Virus 40 infection spreads in old subjects independently from SV40‐contaminated vaccines; (iv) this monkey polyomavirus is also a viral agent in humans.

  April 29, 2016   doi: 10.1002/jcp.25405   open full text
• Modulation of Mammary Stromal Cell Lactate Dynamics by Ambient Glucose and Epithelial Factors.
  Nicolas Tobar, Omar Porras, Patricio C. Smith, L. Felipe Barros, Jorge Martínez.
  Journal of Cellular Physiology. April 28, 2016
  Hyperglycemia is a risk factor for a variety of human cancers. Increased access to glucose and that tumor metabolize glucose by a glycolytic process even in the presence of oxygen (Warburg effect), provide a framework to analyze a particular set of metabolic adaptation mechanisms that may explain this phenomenon. In the present work, using a mammary stromal cell line derived from healthy tissue that was subjected to a long‐term culture in low (5 mM) or high (25 mM) glucose, we analyzed kinetic parameters of lactate transport using a FRET biosensor. Our results indicate that the glucose pre‐culture and soluble epithelial factors constitute a stimulus for lactate stromal production, factors that also modify the kinetic parameters and the monocarboxylate transporters expression in stromal cells. We also observed a vectorial flux of lactate from stroma to epithelial cells in a co‐culture setting and found that the uptake of lactate by epithelial cells correlates with the degree of malignancy. Glucose preconditioning of the stromal cell stimulated epithelial motility. Our findings suggest that lactate generated by stromal cells in the high glucose condition stimulate epithelial migration. Overall, our results support the notion that glucose not only provides a substrate for tumor nutrition but also behaves as a signal promoting malignancy. J. Cell. Physiol. 232: 136–144, 2017. © 2016 Wiley Periodicals, Inc. In a human stromal mammary cell line, glucose pre‐culture and soluble epithelial factors constitute a stimulus for lactate production. In a co‐culture setting, a vectorial flux of lactate from stroma to epithelial cells were stablished. The uptake of lactate by epithelial cells correlates with the degree of malignancy.

  April 28, 2016   doi: 10.1002/jcp.25398   open full text
• Metabolic Adaptation of the Small Intestine to Short‐ and Medium‐Term High‐Fat Diet Exposure.
  Rosmarie Clara, Manuel Schumacher, Deepti Ramachandran, Shahana Fedele, Jean‐Philippe Krieger, Wolfgang Langhans, Abdelhak Mansouri.
  Journal of Cellular Physiology. April 28, 2016
  The small intestine is the main organ involved in the digestion and absorption of nutrients. It is in an ideal position to sense the availability of energy in the lumen in addition to its absorptive function. Consumption of a high‐fat diet (HFD) influences the metabolic characteristics of the small intestine. Therefore, to better understand the metabolic features of the small intestine and their changes in response to dietary fat, we characterized the metabolism of duodenal, jejunal, and hepatic cell lines and assessed the metabolic changes in the enterocytes and the liver after short‐term (3 days) or medium‐term (14 days) HFD feeding in mice. Experiments with immortalized enterocytes indicated a higher glycolytic capacity in the duodenal cell line compared to the other two cell lines, whereas the jejunal cell line exhibited a high oxidative metabolism. Short‐term HFD feeding induced changes in the expression of glucose and lipid metabolism‐related genes in the duodenum and the jejunum of mice, but not in the liver. When focusing on fatty acid oxidation both, short‐ and medium‐term HFD feeding induced an upregulation of 3‐hydroxy‐3‐methylglutaryl‐coenzyme A, the key enzyme of ketogenesis, at the protein level in the intestinal epithelial cells, but not in the liver. These results suggest that HFD feeding induces an early adaptation of the small intestine rather than the liver in response to a substantial fat load. This highlights the importance of the small intestine in the adaptation of the body to the metabolic changes induced by HFD exposure. J. Cell. Physiol. 232: 167–175, 2017. © 2016 Wiley Periodicals, Inc. Characterization of intestinal cell lines from the proximal and distal part of the small intestine, showed a differential metabolism in response to metabolic modulators. Short‐ and medium‐term access to high‐fat‐diet induced the expression of HMG‐CoAS2, the ketogenesis key‐enzyme, specifically in the jejunum but not in the duodenum or in the liver of mice.

  April 28, 2016   doi: 10.1002/jcp.25402   open full text
• Glucocorticoids Hijack Runx2 to Stimulate Wif1 for Suppression of Osteoblast Growth and Differentiation.
  Eri Morimoto, Meng Li, Aysha B. Khalid, Susan A. Krum, Nyam‐Osor Chimge, Baruch Frenkel.
  Journal of Cellular Physiology. April 26, 2016
  Inhibition of Runx2 is one of many mechanisms that suppress bone formation in glucocorticoid (GC)‐induced osteoporosis (GIO). We profiled mRNA expression in ST2/Rx2dox cells after treatment with doxycycline (dox; to induce Runx2) and/or the synthetic GC dexamethasone (dex). As expected, dex typically antagonized Runx2‐driven transcription. Select genes, however, were synergistic stimulated and this was confirmed by RT‐qPCR. Among the genes synergistically stimulated by GCs and Runx2 was Wnt inhibitory Factor 1 (Wif1), and Wif1 protein was readily detectable in medium conditioned by cultures co‐treated with dox and dex, but neither alone. Cooperation between Runx2 and GCs in stimulating Wif1 was also observed in primary preosteoblast cultures. GCs strongly inhibited dox‐driven alkaline phosphatase (ALP) activity in control ST2/Rx2dox cells, but not in cells in which Wif1 was silenced. Unlike its anti‐mitogenic activity in committed osteoblasts, induction of Runx2 transiently increased the percentage of cells in S‐phase and accelerated proliferation in the ST2 mesenchymal pluripotent cell culture model. Furthermore, like the inhibition of Runx2‐driven ALP activity, dex antagonized the transient mitogenic effect of Runx2 in ST2/Rx2dox cultures, and this inhibition eased upon Wif1 silencing. Plausibly, homeostatic feedback loops that rely on Runx2 activation to compensate for bone loss in GIO are thwarted, exacerbating disease progression through stimulation of Wif1. J. Cell. Physiol. 232: 145–153, 2017. © 2016 Wiley Periodicals, Inc. The work by Morimoto et al. demonstrates target gene‐dependent outcomes of the interaction between Runx2 and glucocorticoids (GCs) in preosteoblasts. While Runx2‐driven gene expression is generally antagonized by GCs, a few genes are synergistically stimulated by Runx2 and GCs. One of them, Wif1, potentially thwarts a homeostatic feedback loop and exacerbates GC‐induced osteoporosis.

  April 26, 2016   doi: 10.1002/jcp.25399   open full text
• Runx2/DICER/miRNA Pathway in Regulating Osteogenesis.
  Leilei Zheng, Qisheng Tu, Shu Meng, Lan Zhang, Liming Yu, Jinlin Song, Yun Hu, Lei Sui, Jin Zhang, Michel Dard, Jessica Cheng, Dana Murray, Yin Tang, Jane B. Lian, Gary S. Stein, Jake Chen.
  Journal of Cellular Physiology. April 26, 2016
  DICER is the central enzyme that cleaves precursor microRNAs (miRNAs) into 21–25 nucleotide duplex in cell lineage differentiation, identity, and survival. In the current study, we characterized the specific bone metabolism genes and corresponding miRNAs and found that DICER and Runt‐related transcription factor 2 (Runx2) expressions increased simultaneously during osteogenic differentiation. Luciferase assay showed that Runx2 significantly increased the expression levels of DICER luciferase promoter reporter. Our analysis also revealed weaker DICER expression in embryos of Runx2 knock out mice (Runx2 −/−) compared with that of Runx2 +/− and Runx2 +/+ mice. We further established the calvarial bone critical‐size defect (CSD) mouse model. The bone marrow stromal cells (BMSCs) transfected with siRNA targeting DICER were combined with silk scaffolds and transplanted into calvarial bone CSDs. Five weeks post‐surgery, micro‐CT analysis revealed impaired bone formation, and repairing in calvarial defects with the siRNA targeting DICER group. In conclusion, our results suggest that DICER is specifically regulated by osteogenic master gene Runx2 that binds to the DICER promoter. Consequently, DICER cleaves precursors of miR‐335‐5p and miR‐17‐92 cluster to form mature miRNAs, which target and decrease the Dickkopf‐related protein 1 (DKK1), and proapoptotic factor BIM levels, respectively, leading to an enhanced Wnt/β‐catenin signaling pathway. These intriguing results reveal a central mechanism underlying lineage‐specific regulation by a Runx2/DICER/miRNAs cascade during osteogenic differentiation and bone development. Our study, also suggests a potential application of modulating DICER expression for bone tissue repair and regeneration. J. Cell. Physiol. 232: 182–191, 2017. © 2016 Wiley Periodicals, Inc. These intriguing results reveal a central mechanism underlying lineage‐specific regulation by a Runx2/DICER/miRNAs cascade during osteogenic differentiation and bone development. This cell‐ and development‐specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. Furthermore, our study also suggests a potential application of specifically regulating DICER for bone tissue repair and regeneration.

  April 26, 2016   doi: 10.1002/jcp.25406   open full text
• Atp2c2 Is Transcribed From a Unique Transcriptional Start Site in Mouse Pancreatic Acinar Cells.
  Melissa A. Fenech, Caitlin M. Sullivan, Lucimar T. Ferreira, Rashid Mehmood, William A. MacDonald, Peter B. Stathopulos, Christopher L. Pin.
  Journal of Cellular Physiology. April 21, 2016
  Proper regulation of cytosolic Ca2+ is critical for pancreatic acinar cell function. Disruptions in normal Ca2+ concentrations affect numerous cellular functions and are associated with pancreatitis. Membrane pumps and channels regulate cytosolic Ca2+ homeostasis by promoting rapid Ca2+ movement. Determining how expression of Ca2+ modulators is regulated and the cellular alterations that occur upon changes in expression can provide insight into initiating events of pancreatitis. The goal of this study was to delineate the gene structure and regulation of a novel pancreas‐specific isoform for Secretory Pathway Ca2+ ATPase 2 (termed SPCA2C), which is encoded from the Atp2c2 gene. Using Next Generation Sequencing of RNA (RNA‐seq), chromatin immunoprecipitation for epigenetic modifications and promoter‐reporter assays, a novel transcriptional start site was identified that promotes expression of a transcript containing the last four exons of the Atp2c2 gene (Atp2c2c). This region was enriched for epigenetic marks and pancreatic transcription factors that promote gene activation. Promoter activity for regions upstream of the ATG codon in Atp2c2’s 24th exon was observed in vitro but not in in vivo. Translation from this ATG encodes a protein aligned with the carboxy terminal of SPCA2. Functional analysis in HEK 293A cells indicates a unique role for SPCA2C in increasing cytosolic Ca2+. RNA analysis indicates that the decreased Atp2c2c expression observed early in experimental pancreatitis reflects a global molecular response of acinar cells to reduce cytosolic Ca2+ levels. Combined, these results suggest SPCA2C affects Ca2+ homeostasis in pancreatic acinar cells in a unique fashion relative to other Ca2+ ATPases. J. Cell. Physiol. 231: 2768–2778, 2016. © 2016 Wiley Periodicals, Inc. A novel isoform for SPCA2 (termed SPAC2C) is generated from a unique transcriptional start site within pancreatic acinar cells. Unlike other Ca2+ ATPases, which lower cytosolic Ca2+, SPCA2C increases cytosolic Ca2+, suggesting a novel function for this protein. This is important as SPCA2C is down‐regulated during acinar cell stress and may reflect an effort by these cells to minimize damage associated with dysregulated Ca2+ levels.

  April 21, 2016   doi: 10.1002/jcp.25391   open full text
• SRC Family Kinase Inhibition in Ewing Sarcoma Cells Induces p38 MAP Kinase‐Mediated Cytotoxicity and Reduces Cell Migration.
  Paola Indovina, Nadia Casini, Iris Maria Forte, Tiziana Garofano, Daniele Cesari, Carmelina Antonella Iannuzzi, Leonardo Del Porro, Francesca Pentimalli, Luca Napoliello, Silvia Boffo, Silvia Schenone, Maurizio Botta, Antonio Giordano.
  Journal of Cellular Physiology. April 19, 2016
  Ewing sarcoma (ES) is a highly aggressive bone and soft tissue cancer, representing the second most common primary malignant bone tumor in children and adolescents. Although the development of a multimodal therapy, including both local control (surgery and/or radiation) and systemic multidrug chemotherapy, has determined a significant improvement in survival, patients with metastatic and recurrent disease still face a poor prognosis. Moreover, considering that ES primarily affects young patients, there are concerns about long‐term adverse effects of the therapy. Therefore, more rational strategies, targeting specific molecular alterations underlying ES, are required. Recent studies suggest that SRC family kinases (SFKs), which are aberrantly activated in most cancer types, could represent key therapeutic targets also for ES. Here, we challenged ES cell lines with a recently developed selective SFK inhibitor (a pyrazolo[3,4‐d]pyrimidine derivative, called SI221), which was previously shown to be a valuable proapoptotic agent in other tumor types while not affecting normal cells. We observed that SI221 significantly reduced ES cell viability and proved to be more effective than the well‐known SFK inhibitor PP2. SI221 was able to induce apoptosis in ES cells and also reduced ES cell clonogenic potential. Furthermore, SI221 was also able to reduce ES cell migration. At the molecular level, our data suggest that SFK inhibition through SI221 could reduce ES cell viability at least in part by hindering an SFK‐NOTCH1 receptor‐p38 mitogen‐activated protein kinase (MAPK) axis. Overall, our study suggests a potential application of specific SFK inhibition in ES therapy. J. Cell. Physiol. 232: 129–135, 2017. © 2016 Wiley Periodicals, Inc. In this study, we challenged Ewing sarcoma (ES) cell lines with SI221, a recently developed selective SRC family kinase (SFK) inhibitor. We observed that this compound significantly reduced ES cell viability, clonogenic potential and migration. At the molecular level, our data suggest that SI221 could reduce ES cell viability at least in part by hindering an SFK‐NOTCH1 receptor‐p38 mitogen‐activated protein kinase (MAPK) axis.

  April 19, 2016   doi: 10.1002/jcp.25397   open full text
• MG132 Induces Expression of Monocyte Chemotactic Protein‐Induced Protein 1 in Vascular Smooth Muscle Cells.
  Xi Tan, Jie Gao, Zhan Shi, Shi Tai, Leona Loretta Chan, Yang Yang, Dao‐Quan Peng, Duan‐Fang Liao, Zhi‐Sheng Jiang, Ying‐Zi Chang, Yu Gui, Xi‐Long Zheng.
  Journal of Cellular Physiology. April 14, 2016
  Monocyte chemoattractant protein‐1 (MCP‐1) has been reported to induce the expression of monocyte chemotactic protein‐induced protein 1 (MCPIP1), which undergoes ubiquitination degradation. Therefore, we predict that in vascular smooth muscle (VSMCs), MCPIP1 may be induced by MCP‐1 and undergo degradation, which can be inhibited by the proteasome inhibitor, MG132. Our results showed that treatment of human VSMCs with MCP‐1 did not increase the expression of MCPIP1. Treatment with MG132, however, elevated MCPIP1 protein levels through stimulation of the gene transcription, but not through increasing protein stability. MCPIP1 expression induced by MG132 was inhibited by α‐amanitin inhibition of gene transcription or cycloheximide inhibition of protein synthesis. Our further studies showed that MCPIP1 expression induced by MG132 was inhibited by the inhibitors of AKT and p38 kinase, suggesting a role of the AKT‐p38 pathway in MG132 effects. We also found that treatment with MG132 induces apoptosis, but overexpression of MCPIP1 inhibited bromodeoxyuridine (BrdU) incorporation of human VSMCs without induction of significant apoptosis. In summary, MCPIP1 expression is induced by MG132 likely through activation of the AKT‐p38 pathway. MCPIP1 inhibits SMC proliferation without induction of apoptosis. J. Cell. Physiol. 232: 122–128, 2017. © 2016 Wiley Periodicals, Inc. Monocyte chemoattractant protein‐1 (MCP‐1) has been reported to induce the expression of monocyte chemotactic protein‐induced protein 1 (MCPIP1), which undergoes ubiquitination degradation. In cultured human vascular smooth muscle, MCPIP1 expression is induced by MG132, a proteasome inhibitor, through signal transduction and gene transcription. Overexpression of MCPIP1 results in inhibition of smooth muscle cell proliferation.

  April 14, 2016   doi: 10.1002/jcp.25396   open full text
• l‐glutamine Improves Skeletal Muscle Cell Differentiation and Prevents Myotube Atrophy After Cytokine (TNF‐α) Stress Via Reduced p38 MAPK Signal Transduction.
  Matthew Girven, Hannah F. Dugdale, Daniel J. Owens, David C. Hughes, Claire E. Stewart, Adam P. Sharples.
  Journal of Cellular Physiology. April 14, 2016
  Tumour Necrosis Factor‐Alpha (TNF‐α) is chronically elevated in conditions where skeletal muscle loss occurs. As l‐glutamine can dampen the effects of inflamed environments, we investigated the role of l‐glutamine in both differentiating C2C12 myoblasts and existing myotubes in the absence/presence of TNF‐α (20 ng · ml−1) ± l‐glutamine (20 mM). TNF‐α reduced the proportion of cells in G1 phase, as well as biochemical (CK activity) and morphological differentiation (myotube number), with corresponding reductions in transcript expression of: Myogenin, Igf‐I, and Igfbp5. Furthermore, when administered to mature myotubes, TNF‐α induced myotube loss and atrophy underpinned by reductions in Myogenin, Igf‐I, Igfbp2, and glutamine synthetase and parallel increases in Fox03, Cfos, p53, and Bid gene expression. Investigation of signaling activity suggested that Akt and ERK1/2 were unchanged, JNK increased (non‐significantly) whereas P38 MAPK substantially and significantly increased in both myoblasts and myotubes in the presence of TNF‐α. Importantly, 20 mM l‐glutamine reduced p38 MAPK activity in TNF‐α conditions back to control levels, with a corresponding rescue of myoblast differentiation and a reversal of atrophy in myotubes. l‐glutamine resulted in upregulation of genes associated with growth and survival including; Myogenin, Igf‐Ir, Myhc2 & 7, Tnfsfr1b, Adra1d, and restored atrophic gene expression of Fox03 back to baseline in TNF‐α conditions. In conclusion, l‐glutamine supplementation rescued suppressed muscle cell differentiation and prevented myotube atrophy in an inflamed environment via regulation of p38 MAPK. l‐glutamine administration could represent an important therapeutic strategy for reducing muscle loss in catabolic diseases and inflamed ageing. J. Cell. Physiol. 9999: 231: 2720–2732, 2016. © 2016 Wiley Periodicals, Inc. l‐glutamine supplementation rescued suppressed muscle cell differentiation and prevented myotube atrophy in an inflamed cytokine environment (TNF‐α) via regulation of p38 MAPK singaling. l‐Glutamine supplementation evoked reductions in p38 MAPK following extensive increases after TNF‐α administration. This was associated with downstream regulation of gene transcription associated with growth and survival including increased; Myogenin, Igf‐Ir, Myhc2 & 7, Tnfsfr1b, Adra1d, and reduced atrophic gene expression of Fox03 back to baseline in TNF‐α conditions. l‐glutamine administration could represent an important therapeutic strategy for reducing muscle loss in catabolic diseases and inflamed ageing.

  April 14, 2016   doi: 10.1002/jcp.25380   open full text
• Effects of 1,25(OH)2D3 and 25(OH)D3 on C2C12 Myoblast Proliferation, Differentiation, and Myotube Hypertrophy.
  K. van der Meijden, N. Bravenboer, N.F. Dirks, A.C. Heijboer, M. den Heijer, G.M.J. de Wit, C. Offringa, P. Lips, R.T. Jaspers.
  Journal of Cellular Physiology. April 14, 2016
  An adequate vitamin D status is essential to optimize muscle strength. However, whether vitamin D directly reduces muscle fiber atrophy or stimulates muscle fiber hypertrophy remains subject of debate. A mechanism that may affect the role of vitamin D in the regulation of muscle fiber size is the local conversion of 25(OH)D to 1,25(OH)2D by 1α‐hydroxylase. Therefore, we investigated in a murine C2C12 myoblast culture whether both 1,25(OH)2D3 and 25(OH)D3 affect myoblast proliferation, differentiation, and myotube size and whether these cells are able to metabolize 25(OH)D3 and 1,25(OH)2D3. We show that myoblasts not only responded to 1,25(OH)2D3, but also to the precursor 25(OH)D3 by increasing their VDR mRNA expression and reducing their proliferation. In differentiating myoblasts and myotubes 1,25(OH)2D3 as well as 25(OH)D3 stimulated VDR mRNA expression and in myotubes 1,25(OH)2D3 also stimulated MHC mRNA expression. However, this occurred without notable effects on myotube size. Moreover, no effects on the Akt/mTOR signaling pathway as well as MyoD and myogenin mRNA levels were observed. Interestingly, both myoblasts and myotubes expressed CYP27B1 and CYP24 mRNA which are required for vitamin D3 metabolism. Although 1α‐hydroxylase activity could not be shown in myotubes, after treatment with 1,25(OH)2D3 or 25(OH)D3 myotubes showed strongly elevated CYP24 mRNA levels compared to untreated cells. Moreover, myotubes were able to convert 25(OH)D3 to 24R,25(OH)2D3 which may play a role in myoblast proliferation and differentiation. These data suggest that skeletal muscle is not only a direct target for vitamin D3 metabolites, but is also able to metabolize 25(OH)D3 and 1,25(OH)2D3. J. Cell. Physiol. 231: 2517–2528, 2016. © 2016 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. In differentiating C2C12 myoblasts and myotubes, 1,25(OH)2D3 as well as 25(OH)D3 stimulate VDR mRNA expression and in myotubes 1,25(OH)2D3 also stimulates MHC mRNA expression, however, this occurs without notable effects on expression of myogenic regulatory factors, the Akt/mTOR signaling pathway and myotube size. Interestingly, both differentiating myoblasts and myotubes express CYP27B1 and CYP24 mRNA and myotubes are able to synthesize 24R,25(OH)2D3 from 25(OH)D3. These data suggest that skeletal muscle is not only a direct target for vitamin D3 metabolites, but is also able to metabolize 25(OH)D3 and 1,25(OH)2D3.

  April 14, 2016   doi: 10.1002/jcp.25388   open full text
• XPO1 Inhibition Preferentially Disrupts the 3D Nuclear Organization of Telomeres in Tumor Cells.
  Cheryl Taylor‐Kashton, Daniel Lichtensztejn, Erkan Baloglu, William Senapedis, Sharon Shacham, Michael G. Kauffman, Rami Kotb, Sabine Mai.
  Journal of Cellular Physiology. April 08, 2016
  Previous work has shown that the three‐dimensional (3D) nuclear organization of telomeres is altered in cancer cells and the degree of alterations coincides with aggressiveness of disease. Nuclear pores are essential for spatial genome organization and gene regulation and XPO1 (exportin 1/CRM1) is the key nuclear export protein. The Selective Inhibitor of Nuclear Export (SINE) compounds developed by Karyopharm Therapeutics (KPT‐185, KPT‐330/selinexor, and KPT‐8602) inhibit XPO1 nuclear export function. In this study, we investigated whether XPO1 inhibition has downstream effects on the 3D nuclear organization of the genome. This was assessed by measuring the 3D telomeric architecture of normal and tumor cells in vitro and ex vivo. Our data demonstrate for the first time a rapid and preferential disruption of the 3D nuclear organization of telomeres in tumor cell lines and in primary cells ex vivo derived from treatment‐naïve newly diagnosed multiple myeloma patients. Normal primary cells in culture as well as healthy lymphocyte control cells from the same patients were minimally affected. Using both lymphoid and non‐lymphoid tumor cell lines, we found that the downstream effects on the 3D nuclear telomere structure are independent of tumor type. We conclude that the 3D nuclear organization of telomeres is a sensitive indicator of cellular response when treated with XPO1 inhibitors. J. Cell. Physiol. 231: 2711–2719, 2016. © 2016 Wiley Periodicals, Inc. This study shows that XPO1 inhibition directly disrupts the 3D nuclear telomere organization of tumor cell lines and of primary tumor cells (multiple myeloma) from treatment naive patients. Normal cells are only minimally afffected.

  April 08, 2016   doi: 10.1002/jcp.25378   open full text
• Quercetin Partially Preserves Development of Osteoblast Phenotype in Fetal Rat Calvaria Cells in an Oxidative Stress Environment.
  Jonathan G. Messer, Stephanie La, Robin G. Hopkins, Deborah E. Kipp.
  Journal of Cellular Physiology. April 08, 2016
  Studies are needed to improve understanding of the osteoblast antioxidant response, and the balance between oxidative homeostasis and osteoblast differentiation. The flavonol quercetin aglycone (QRC) up‐regulates the osteoblast antioxidant response in vitro without suppressing osteoblast phenotype, suggesting that QRC may preserve osteoblast phenotypic development in cells subsequently exposed to oxidative stress, which suppresses osteoblast differentiation. The aims of this study were to assess the extent that QRC pretreatment preserved development of the osteoblast phenotype in cells subsequently cultured with hydrogen peroxide, an oxidative stressor, and to characterize alterations in the osteoblast antioxidant response and in key antioxidant signaling pathways. We hypothesized that pretreatment with QRC would preserve phenotypic development after hydrogen peroxide treatment, suppress the hydrogen peroxide‐induced antioxidant response, and that the antioxidant response would involve alterations in Nrf2 and ERK1/2 signaling. Results showed that treating fetal rat calvarial osteoblasts for 4 days (D5‐9) with 300 μM hydrogen peroxide resulted in fewer alkaline phosphatase‐positive cells and mineralized nodules, altered cell morphology, and significantly lower osteoblast phenotypic gene expression (P < 0.05). This suppression was partially blocked when cells were pretreated 12 h with 20 μM QRC. Hydrogen peroxide also produced sustained up‐regulation of heme oxygenase‐1 (HO‐1) and γ‐glutamate cysteine ligase catalytic subunit (GCLC), which was partially blocked in hydrogen peroxide‐treated cells that first received QRC pretreatment. The alterations in the antioxidant stress response coincided with alterations in phosphorylated ERK1/2, but not Nrf2. These results suggest that QRC suppresses hydrogen peroxide‐induced activation of the antioxidant response, and partially preserves osteoblast phenotypic development. J. Cell. Physiol. 231: 2779–2788, 2016. © 2016 Wiley Periodicals, Inc. Oxidative stress induces the antioxidant response, and at the same time suppresses the development of osteoblasts in culture. However, briefly exposing cells to an antioxidant, quercetin aglycone, before exposure to oxidative stress appears to reverse these effects. This study provides evidence supporting the hypothesis that the antioxidant stress response and osteoblast phenotypic development pathways are reciprocally linked.

  April 08, 2016   doi: 10.1002/jcp.25392   open full text
• Cannabinoid Receptor 1 Mediates Homing of Bone Marrow‐Derived Mesenchymal Stem Cells Triggered by Chronic Liver Injury.
  Lin Wang, Le Yang, Lei Tian, Ping Mai, Shuangshuang Jia, Lin Yang, Liying Li.
  Journal of Cellular Physiology. April 08, 2016
  Cannabinoid receptors (CBs) have been implicated in the pathogenesis of various liver diseases, including liver fibrosis. Our previous studies have demonstrated that after liver injury, mouse bone marrow‐derived mesenchymal stem cells (BMSCs) can migrate to the injured liver and differentiate to myofibroblasts, contributing to hepatic fibrogenesis. However, the role of CBs in the homing of BMSCs in liver injury is yet unclear. In this study, we found that both CB1 and CB2 were expressed in BMSCs. Migration assays were performed by transwell chambers. CB1 agonist ACEA promoted the migration of BMSCs, but CB2 agonist JWH133 had no effect. Pharmacological or genetic ablation of CB1 reduced ACEA‐induced migration, whereas CB2 did not. Moreover, activation of CB1 increased active GTP‐bound Rac1, RhoA, and Cdc42 protein levels. The elevated GTP‐bound Rac1 and RhoA protein levels were decreased by CB1 antagonist AM281 treatment, but not Cdc42. In addition, ACEA‐induced migration was suppressed by NSC23766 (Rac1 inhibitor) or C3 transferase (RhoA inhibitor), whereas MLS‐573151 (Cdc42 inhibitor) had no effect. Consistent with these data, Rac1 or RhoA knock‐down significantly blocked CB1‐mediated migration. Meanwhile, CB1‐mediated migration was associated with cytoskeletal remodeling. In vivo, administration of CB1 antagonist AM281 markedly inhibited the recruitment of BMSCs to the injured liver using fluorescence‐activated cell sorting. Furthermore, blockade of CB1 significantly attenuated liver fibrosis. In conclusion, our results suggest that CB1 plays a crucial role in liver fibrosis through mediating the homing of BMSCs to damaged liver, which may provide new insight into the pathogenesis and treatment of liver fibrosis. J. Cell. Physiol. 232: 110–121, 2017. © 2016 Wiley Periodicals, Inc. The present study aims to evaluate the impact of Cannabinoid receptors (CBs) on the migration of BMSCs in liver injury and investigate the mechanisms involved. We found that BMSCs expressed CB1 and CB2. CB1 rather than CB2 mediated the migration of BMSCs via the Rac1 and RhoA pathways. And CB1‐mediated migration was associated with cytoskeletal remodeling. Furthermore, pharmacological blockade of CB1 markedly reduced the recruitment of BMSCs to injured liver and attenuated liver fibrosis. These results may contribute to the better understanding of the involvement of CB1 in liver injury and open new perspectives for development of potential therapeutics for liver disease.

  April 08, 2016   doi: 10.1002/jcp.25395   open full text
• Enhancement of Lytic Activity by Leptin Is Independent From Lipid Rafts in Murine Primary Splenocytes.
  Aurore Collin, Audrey Noacco, Jérémie Talvas, Florence Caldefie‐Chézet, Marie‐Paule Vasson, Marie‐Chantal Farges.
  Journal of Cellular Physiology. April 07, 2016
  Leptin, a pleiotropic adipokine, is known as a regulator of food intake, but it is also involved in inflammation, immunity, cell proliferation, and survival. Leptin receptor is integrated inside cholesterol‐rich microdomains called lipid rafts, which, if disrupted or destroyed, could lead to a perturbation of lytic mechanism. Previous studies also reported that leptin could induce membrane remodeling. In this context, we studied the effect of membrane remodeling in lytic activity modulation induced by leptin. Thus, primary mouse splenocytes were incubated with methyl‐β‐cyclodextrin (β‐MCD), a lipid rafts disrupting agent, cholesterol, a major component of cell membranes, or ursodeoxycholic acid (UDCA), a membrane stabilizer agent for 1 h. These treatments were followed by splenocyte incubation with leptin (absence, 10 and 100 ng/ml). Unlike β‐MCD or cholesterol, UDCA was able to block leptin lytic induction. This result suggests that leptin increased the lytic activity of primary spleen cells against syngenic EO771 mammary cancer cells independently from lipid rafts but may involve membrane fluidity. Furthermore, natural killer cells were shown to be involved in the splenocyte lytic activity. To our knowledge it is the first publication in primary culture that provides the link between leptin lytic modulation and membrane remodeling. J. Cell. Physiol. 232: 101–109, 2017. © 2016 Wiley Periodicals, Inc. This article suggests that leptin increased the lytic activity of primary spleen cells against syngenic EO771 mammary cancer cells independently from lipid rafts but may involve membrane fluidity. Furthermore, natural killer cells were shown to be involved in the splenocyte lytic activity. To our knowledge it is the first publication in primary culture that provides the link between leptin lytic modulation and membrane remodeling.

  April 07, 2016   doi: 10.1002/jcp.25394   open full text
• Serum From Advanced Heart Failure Patients Promotes Angiogenic Sprouting and Affects the Notch Pathway in Human Endothelial Cells.
  Micaela Pannella, Cristiana Caliceti, Francesca Fortini, Giorgio Aquila, Francesco Vieceli Dalla Sega, Antonio Pannuti, Cinzia Fortini, Marco Bruno Morelli, Alessandro Fucili, Gloria Francolini, Rebecca Voltan, Paola Secchiero, Giovanni Dinelli, Emanuela Leoncini, Manuela Ferracin, Silvana Hrelia, Lucio Miele, Paola Rizzo.
  Journal of Cellular Physiology. April 04, 2016
  It is unknown whether components present in heart failure (HF) patients’ serum provide an angiogenic stimulus. We sought to determine whether serum from HF patients affects angiogenesis and its major modulator, the Notch pathway, in human umbilical vein endothelial cells (HUVECs). In cells treated with serum from healthy subjects or from patients at different HF stage we determined: (1) Sprouting angiogenesis, by measuring cells network (closed tubes) in collagen gel. (2) Protein levels of Notch receptors 1, 2, 4, and ligands Jagged1, Delta‐like4. We found a higher number of closed tubes in HUVECs treated with advanced HF patients serum in comparison with cells treated with serum from mild HF patients or controls. Furthermore, as indicated by the reduction of the active form of Notch4 (N4IC) and of Jagged1, advanced HF patients serum inhibited Notch signalling in HUVECs in comparison with mild HF patients’ serum and controls. The circulating levels of NT‐proBNP (N‐terminal of the pro‐hormone brain natriuretic peptide), a marker for the detection and evalutation of HF, were positively correlated with the number of closed tubes (r = 0.485) and negatively with Notch4IC and Jagged1 levels in sera‐treated cells (r = −0.526 and r = −0.604, respectively). In conclusion, we found that sera from advanced HF patients promote sprouting angiogenesis and dysregulate Notch signaling in HUVECs. Our study provides in vitro evidence of an angiogenic stimulus arising during HF progression and suggests a role for the Notch pathway in it. J. Cell. Physiol. 231: 2700–2710, 2016. © 2016 Wiley Periodicals, Inc. It is unknown whether components present in heart failure (HF) patients’ serum provide an angiogenic stimulus. We show that sera from advanced HF patients promote sprouting angiogenesis and dysregulate Notch signaling in human umbilical vein endothelial cells. Our study provides in vitro evidence of an angiogenic stimulus arising during HF progression and suggests a role for the Notch pathway in it.

  April 04, 2016   doi: 10.1002/jcp.25373   open full text
• Kynurenine Modulates MMP‐1 and Type‐I Collagen Expression Via Aryl Hydrocarbon Receptor Activation in Dermal Fibroblasts.
  Malihe‐Sadat Poormasjedi‐Meibod, Sanam Salimi Elizei, Victor Leung, Reza Baradar Jalili, Frank Ko, Aziz Ghahary.
  Journal of Cellular Physiology. April 04, 2016
  Dermal fibrosis is characterized by a high deposition of extracellular matrix (ECM) and tissue cellularity. Unfortunately all means of treating this condition are unsatisfactory. We have previously reported the anti‐fibrotic effects of Kynurenine (Kyn), a tryptophan metabolite, in fibrotic rabbit ear model. Here, we report the mechanism by which Kyn modulates the expression of key ECM components in dermal fibroblasts. The results showed that Kyn activates aryl hydrocarbon receptor (AHR) nuclear translocation and up‐regulates cytochrome‐P450 (CYP1A‐1) expression, the AHR target gene. A specific AHR antagonist, 6,2′,4′‐trimethoxyflavone, inhibited the Kyn‐dependent modulation of CYP1A‐1, MMP‐1, and type‐I collagen expression. Establishing the anti‐fibrogenic effect of Kyn and its mechanism of action, we then developed nano‐fibrous Kyn slow‐releasing dressings and examined their anti‐fibrotic efficacy in vitro and in a rat model. Our results showed the feasibility of incorporating Kyn into PVA/PLGA nanofibers, prolonging the Kyn release up to 4 days tested. Application of medicated‐dressings significantly improved the dermal fibrosis indicated by MMP‐1 induction, alpha‐smooth muscle actin and type‐I collagen suppression, and reduced tissue cellularity, T‐cells and myofibroblasts. This study clarifies the mechanism by which Kyn modulates ECM expression and reports the development of a new slow‐releasing anti‐fibrogenic dressing. J. Cell. Physiol. 231: 2749–2760, 2016. © 2016 Wiley Periodicals, Inc. Kynurenine modulates the expression of collagen and collagenase (MMP‐1) via aryl hydrocarbon receptor signaling pathway. The medicated nanofibrous wound dressings, developed in this study, can be used as an effective slow releasing drug delivery system in order to prevent the post‐burn and pots‐surgical hypertrophic scar formation.

  April 04, 2016   doi: 10.1002/jcp.25383   open full text
• A Novel Biological Role of α‐Mangostin in Modulating Inflammatory Response Through the Activation of SIRT‐1 Signaling Pathway.
  Sara Franceschelli, Mirko Pesce, Alessio Ferrone, Antonia Patruno, Livia Pasqualone, Giuseppe Carlucci, Vincenzo Ferrone, Maura Carlucci, Maria Anna de Lutiis, Alfredo Grilli, Mario Felaco, Lorenza Speranza.
  Journal of Cellular Physiology. April 04, 2016
  Several studies have shown that xanthones obtained from Garcinia Mangostana (GM) have remarkable biological activities. α‐mangostin (α‐MG) is the main constituent of the fruit hull of the GM. Several findings have suggested that SIRT‐1, a nuclear histone deacetylase, could influence cellular function by the inhibition of NF‐kB signaling. ROS can inhibit SIRT‐1 activity by initiating oxidative modifications on its cysteine residues, and suppression of SIRT‐1 enhances the NF‐κB signaling resulting in inflammatory responses. The goals of the present study were to evaluate the quantity of α‐MG in the methanolic extract of GM (Vithagroup Spa) and to investigate the activity of this xanthone in U937 cell line and in human monocytes from responsive to inflammatory insult analyzing the possible changes on the activation of SIRT‐1 protein via NF‐Kb. Cells were treated with the methanolic extract of GM and/or LPS. The chromatographic separation of α‐MG was performed by an HPLC analysis. EX 527, a specific SIRT‐1 inhibitor, was used to determine if SIRT‐1/NfkB signaling pathway might be involved in α‐MG action on cells. Our results show that α‐MG inhibits p65 acetylation and down‐regulates the pro‐inflammatory gene products as COX‐2, iNOS via SIRT‐1 activation. Cells treated with EX 527 showed an up‐regulation of NFkB acetylation and an over expression of inducible enzymes and their product of catalysis (NO and PGE2). These results suggest that α‐MG may be useful for the development of alternative pharmacological strategies aimed at reducing the inflammatory process. J. Cell. Physiol. 231: 2439–2451, 2016. © 2016 Wiley Periodicals, Inc.

  April 04, 2016   doi: 10.1002/jcp.25348   open full text
• Phospholipidomic Profile Variation on THP‐1 Cells Exposed to Skin or Respiratory Sensitizers and Respiratory Irritant.
  João D. Martins, Elisabete A. Maciel, Ana Silva, Isabel Ferreira, Fernando Ricardo, Pedro Domingues, Bruno M. Neves, Maria Rosário M. Domingues, Maria Teresa Cruz.
  Journal of Cellular Physiology. April 04, 2016
  Occupational exposure to low molecular weight reactive chemicals often leads to development of allergic reactions such as allergic contact dermatitis and respiratory allergies. Further insights into the interaction of these chemicals with physiopathological relevant cellular models might provide the foundations for novel non‐animal approaches to safety assessment. In this work we used the human THP‐1 cell line to determine phospholipidome changes induced by the skin sensitizer 1‐fluoro‐2,4‐dinitrobenzene (DNFB), the respiratory allergen hexamethylene diisocyanate (HDI), and the irritant methyl salicylate (MESA). We detected that these chemicals differently induce lipid peroxidation and modulate THP‐1 IL‐1β, IL‐12B, IL‐8, CD86, and HMOX1 transcription. Decreased phosphatidylethanolamine content was detected in cells exposed to MESA, while profound alterations in the relative abundance of cardiolipin species were observed in cells exposed to DNFB. All chemicals tested induced a decrease in the relative abundance of plasmanyl phosphatidylcholine species PC (O‐16:0e/18:1) and phosphatidylinositol species PI (34:1), while increasing PI (38:4). An increased abundance of oleic acid was observed in the phospholipids of cells exposed to DNFB while a decreased abundance of palmitic acid was detected in cells treated with MESA or DNFB. We conclude that both specific and common alterations at phospholipidome levels are triggered by the different chemicals, while not allowing a complete distinction between them using a Canonical Analysis of Principal Coordinates (CAP). The common effects observed at phospholipids level with all the chemicals tested might be related to unspecific cell cytotoxic mechanisms that nevertheless may contribute to the elicitation of specific immune responses. J. Cell. Physiol. 231: 2639–2651, 2016. © 2016 Wiley Periodicals, Inc. The skin sensitizer 1‐fluoro‐2,4‐dinitrobenzene (DNFB), the respiratory allergen hexamethylene diisocyanate (HDI), and the irritant methyl salicylate (MESA) trigger both specific and common alterations at THP‐1 phospholipidome level. To some extent these alterations are dependent on the oxidative capacity of the chemicals and may be important in shaping THP‐1 cells’ maturation.

  April 04, 2016   doi: 10.1002/jcp.25365   open full text
• The Impact of Epigenetics on Mesenchymal Stem Cell Biology.
  Yusuf Ozkul, Umberto Galderisi.
  Journal of Cellular Physiology. April 04, 2016
  Changes in epigenetic marks are known to be important regulatory factors in stem cell fate determination and differentiation. In the past years, the investigation of the epigenetic regulation of stem cell biology has largely focused on embryonic stem cells (ESCs). Contrarily, less is known about the epigenetic control of gene expression during differentiation of adult stem cells (AdSCs). Among AdSCs, mesenchymal stem cells (MSCs) are the most investigated stem cell population because of their enormous potential for therapeutic applications in regenerative medicine and tissue engineering. In this review, we analyze the main studies addressing the epigenetic changes in MSC landscape during in vitro cultivation and replicative senescence, as well as follow osteocyte, chondrocyte, and adipocyte differentiation. In these studies, histone acetylation, DNA methylation, and miRNA expression are among the most investigated phenomena. We describe also epigenetic changes that are associated with in vitro MSC trans‐differentiation. Although at the at initial stage, the epigenetics of MSCs promise to have profound implications for stem cell basic and applied research. J. Cell. Physiol. 231: 2393–2401, 2016. © 2016 Wiley Periodicals, Inc. Epigenetic marks are known to be important regulatory factors in stem cell fate determination and differentiation. In this paper, we reviewed studies on epigenetic changes that govern mesenchymal stem cell biology.

  April 04, 2016   doi: 10.1002/jcp.25371   open full text
• E‐Cadherin Facilitates Protein Kinase D1 Activation and Subcellular Localization.
  Zhuo Li, Chuanyou Zhang, Li Chen, Guosheng Li, Ling Qu, K.C. Balaji, Cheng Du.
  Journal of Cellular Physiology. April 01, 2016
  Protein kinase D 1 (PKD1) is a serine/threonine kinase implicated in the regulation of diverse cellular functions including cell growth, differentiation, adhesion and motility. The current model for PKD1 activation involves diacylglycerol (DAG) binding to the C1 domain of PKD1 which results in the translocation of PKD1 to subcellular membranes where PKD1 is phosphorylated and activated by protein kinase C (PKC). In this study, we have identified a novel regulation of PKD1 activation. The epithelial cell membrane protein E‐cadherin physically binds to PKD1 which leads to a subcellular redistribution of PKD1. Furthermore, artificial targeting of PKD1 to the membrane leads to PKD1 activation in a PKC‐independent manner, indicating that membrane attachment is sufficient enough to activate PKD1. The presence of E‐cadherin dynamically regulates PKD1 activation by Bryostatin 1, a potent activator of PKD1, and its substrate phosphorylation specificity, implying a loss of E‐cadherin during cancer metastasis could cause the re‐distribution PKD1 and re‐wiring of PKD1 signaling for distinct functions. The knocking down of PKD1 in lung epithelial cell line A549 results in an epithelial to mesenchymal transition with changes in biomarker expression, cell migration and drug resistance. These results extend our previous understanding of PKD1 regulation and E‐cadherin signaling functions and may help to explain the diversified functions of PKD1 in various cells. J. Cell. Physiol. 231: 2741–2748, 2016. © 2016 Wiley Periodicals, Inc. E‐cadherin intracellular domain directly binds to PKD1 C1 domain. This interesction regulates PKD1 subcellular localization and activation.

  April 01, 2016   doi: 10.1002/jcp.25382   open full text
• CRISPR‐Cas‐Assisted Multiplexing (CAM): Simple Same‐Day Multi‐Locus Engineering in Yeast.
  Jessica M. Walter, Sunil S. Chandran, Andrew A. Horwitz.
  Journal of Cellular Physiology. March 31, 2016
  Demands on the industrial and academic yeast strain engineer have increased significantly in the era of synthetic biology. Installing complex biosynthetic pathways and combining point mutations are tedious and time‐consuming using traditional methods. With multiplex engineering tools, these tasks can be completed in a single step, typically achieving up to sixfold compression in strain engineering timelines. To capitalize on this potential, a variety of yeast CRISPR‐Cas methods have been developed, differing largely in how the guide RNA (gRNA) reagents that direct the Cas9 nuclease are delivered. However, in nearly all reported protocols, the time savings of multiplexing is offset by multiple days of cloning to prepare the required reagents. Here, we discuss the advantages and opportunities of CRISPR‐Cas‐assisted multiplexing (CAM), a same‐day, cloning‐free method for multi‐locus engineering in yeast. J. Cell. Physiol. 231: 2563–2569, 2016. © 2016 Wiley Periodicals, Inc. Installing complex biosynthetic pathways and combining point mutations in yeast are tedious and time‐consuming using traditional methods. Here, we discuss the advantages and opportunities of CRISPR‐Cas‐assisted multiplexing (CAM), a same‐day, cloning‐free method for multi‐locus engineering.

  March 31, 2016   doi: 10.1002/jcp.25375   open full text
• TNF‐α Induces Caspase‐1 Activation Independently of Simultaneously Induced NLRP3 in 3T3‐L1 Cells.
  Mana Furuoka, Kei‐Ichi Ozaki, Daichi Sadatomi, Sayaka Mamiya, Tomo Yonezawa, Susumu Tanimura, Kohsuke Takeda.
  Journal of Cellular Physiology. March 30, 2016
  The intracellular cysteine protease caspase‐1 is critically involved in obesity‐induced inflammation in adipose tissue. A substantial body of evidence from immune cells, such as macrophages, has shown that caspase‐1 activation depends largely on a protein complex, called the NLRP3 inflammasome, which consists of the NOD‐like receptor (NLR) family protein NLRP3, the adaptor protein ASC, and caspase‐1 itself. However, it is not fully understood how caspase‐1 activation is regulated within adipocytes upon inflammatory stimuli. In this study, we show that TNF‐α‐induced activation of caspase‐1 is accompanied by robust induction of NLRP3 in 3T3‐L1 adipocytes but that caspase‐1 activation may not depend on the NLRP3 inflammasome. Treatment of 3T3‐L1 cells with TNF‐α induced mRNA expression and activation of caspase‐1. Although the basal expression of NLRP3 and ASC was undetectable in unstimulated cells, TNF‐α strongly induced NLRP3 expression but did not induce ASC expression. Interestingly, inhibitors of the ERK MAP kinase pathway strongly suppressed NLRP3 expression but did not suppress the expression and activation of caspase‐1 induced by TNF‐α, suggesting that NLRP3 is dispensable for TNF‐α‐induced caspase‐1 activation. Moreover, we did not detect the basal and TNF‐α‐induced expression of other NLR proteins (NLRP1a, NLRP1b, and NLRC4), which do not necessarily require ASC for caspase‐1 activation. These results suggest that TNF‐α induces caspase‐1 activation in an inflammasome‐independent manner in 3T3‐L1 cells and that the ERK‐dependent expression of NLRP3 may play a role independently of its canonical role as a component of inflammasomes. J. Cell. Physiol. 231: 2761–2767, 2016. © 2016 Wiley Periodicals, Inc. In this study, we propose that TNF‐α induces caspase‐1 activation in an inflammasome‐independent manner in 3T3‐L1 cells and that the ERK‐dependent expression of NLRP3 may play a role independently of its canonical role as a component of inflammasomes.

  March 30, 2016   doi: 10.1002/jcp.25385   open full text
• Stimulatory Effects of Coumestrol on Embryonic and Fetal Development Through AKT and ERK1/2 MAPK Signal Transduction.
  Whasun Lim, Gwonhwa Song.
  Journal of Cellular Physiology. March 30, 2016
  Successful establishment of pregnancy is required for fetal‐maternal interactions regulating implantation, embryonic development and placentation. A uterine environment with insufficient growth factors and nutrients increases the incidence of intrauterine growth restriction (IUGR) leading to an impaired uterine environment. In the present study, we demonstrated the effects of the phytoestrogen coumestrol on conceptus development in the pig that is regarded as an excellent biomedical animal model for research on IUGR. Results of this study indicated that coumestrol induced migration of porcine trophectoderm (pTr) cells in a concentration‐dependent manner. In response to coumestrol, the phosphorylation of AKT, P70S6K, S6, ERK1/2 MAPK, and P90RSK proteins were activated in pTr cells and ERK1/2 MAPK and P90RSK phosphorylation was prolonged for a longer period than for the other proteins. To identify the signal transduction pathway induced by coumestrol, pharmacological inhibitors U0126 (an ERK1/2 inhibitor) and LY294002 (a PI3K inhibitor) were used to pretreat pTr cells. The results showed that coumestrol‐induced phosphorylation of ERK1/2 MAPK and P90RSK was blocked by U0126. In addition, the increased phosphorylation in response to coumestrol was completely inhibited following pre‐treatment incubation of pTr cells in the presence of LY294002 and U0126. Furthermore, these two inhibitors suppressed the ability of coumestrol to induce migration of pTr cells. Collectively, these findings suggest that coumestrol affects embryonic development through activation of the PI3K/AKT and ERK1/2 MAPK cell signal transduction pathways and improvement in the uterine environment through coumestrol supplementation may provide beneficial effects of enhancing embryonic and fetal survival and development. J. Cell. Physiol. 231: 2733–2740, 2016. © 2016 Wiley Periodicals, Inc. Coumestrol stimulates migration of trophectoderm cells which is required for elongation and development of porcine conceptuses. Coumestrol affects embryonic development through activation of the PI3K/AKT and ERK1/2 MAPK cell signal transduction pathways.

  March 30, 2016   doi: 10.1002/jcp.25381   open full text
• Priming Endothelial Cells With a Melanoma‐Derived Extracellular Matrix Triggers the Activation of αvβ3/VEGFR2 Axis.
  Edward Helal‐Neto, Renata M. Brandão‐Costa, Roberta Saldanha‐Gama, Cristiane Ribeiro‐Pereira, Victor Midlej, Marlene Benchimol, Verônica Morandi, Christina Barja‐Fidalgo.
  Journal of Cellular Physiology. March 30, 2016
  The unique composition of tumor‐produced extracellular matrix (ECM) can be a determining factor in changing the profile of endothelial cells in the tumor microenvironment. As the main receptor for ECM proteins, integrins can activate a series of signaling pathways related to cell adhesion, migration, and differentiation of endothelial cells that interact with ECM proteins. We studied the direct impact of the decellularized ECM produced by a highly metastatic human melanoma cell line (MV3) on the activation of endothelial cells and identified the intracellular signaling pathways associated with cell differentiation. Our data show that compared to the ECM derived from a human melanocyte cell line (NGM‐ECM), ECM produced by a melanoma cell line (MV3‐ECM) is considerably different in ultrastructural organization and composition and possesses a higher content of tenascin‐C and laminin and a lower expression of fibronectin. When cultured directly on MV3‐ECM, endothelial cells change morphology and show increased adhesion, migration, proliferation, and tubulogenesis. Interaction of endothelial cells with MV3‐ECM induces the activation of integrin signaling, increasing FAK phosphorylation and its association with Src, which activates VEGFR2, potentiating the receptor response to VEGF. The blockage of αvβ3 integrin inhibited the FAK‐Src association and VEGFR activation, thus reducing tubulogenesis. Together, our data suggest that the interaction of endothelial cells with the melanoma‐ECM triggers integrin‐dependent signaling, leading to Src pathway activation that may potentiate VEGFR2 activation and up‐regulate angiogenesis. J. Cell. Physiol. 231: 2464–2473, 2016. © 2016 Wiley Periodicals, Inc.

  March 30, 2016   doi: 10.1002/jcp.25358   open full text
• MMP‐8 Is Critical for Dexamethasone Therapy in Alkali‐Burned Corneas Under Dry Eye Conditions.
  Fang Bian, Changjun Wang, Johanna Tukler‐Henriksson, Stephen C. Pflugfelder, Caterina Camodeca, Elisa Nuti, Armando Rossello, De‐Quan Li, Cintia S. de Paiva.
  Journal of Cellular Physiology. March 28, 2016
  Our previous studies have shown that Dexamethasone (Dex) reduced the expression of matrix‐metalloproteinases (MMPs ‐1,‐3,‐9,‐13), IL‐1β and IL‐6, while it significantly increased MMP‐8 mRNA transcripts in a concomitant dry eye and corneal alkali burn murine model (CM). To investigate if MMP‐8 induction is responsible for some of the protective effects of Dex in CM, MMP‐8 knock out mice (MMP‐8KO) were subjected to the CM for 2 or 5 days and topically treated either with 2 μl of 0.1% Dexamethasone (Dex), or saline QID. A separate group of C57BL/6 mice were topically treated with Dex or BSS and received either 100 nM CAM12 (MMP‐8 inhibitor) or vehicle IP, QD. Here we demonstrate that topical Dex treated MMP‐8KO mice subjected to CM showed reduced corneal clarity, increased expression of inflammatory mediators (IL‐6, CXCL1, and MMP‐1 mRNA) and increased neutrophil infiltration at 2D and 5D compared to Dex treated WT mice. C57BL/6 mice topically treated with Dex and CAM12 IP recapitulated findings seen with MMP‐8KO mice. These results suggest that some of the anti‐inflammatory effects of Dex are mediated through increased MMP‐8 expression. J. Cell. Physiol. 231: 2506–2516, 2016. © 2016 Wiley Periodicals, Inc.

  March 28, 2016   doi: 10.1002/jcp.25364   open full text
• Role of PTHrP(1‐34) Pulse Frequency Versus Pulse Duration to Enhance Mesenchymal Stromal Cell Chondrogenesis.
  Jennifer Fischer, Marlen Ortel, Sebastien Hagmann, Andreas Hoeflich, Wiltrud Richter.
  Journal of Cellular Physiology. March 21, 2016
  Generation of phenotypically stable, articular chondrocytes from mesenchymal stromal cells (MSCs) is still an unaccomplished task, with formation of abundant, hyaline extracellular matrix, and avoidance of hypertrophy being prime challenges. We recently demonstrated that parathyroid hormone‐related protein (PTHrP) is a promising factor to direct chondrogenesis of MSCs towards an articular phenotype, since intermittent PTHrP application stimulated cartilage matrix production and reduced undesired hypertrophy. We here investigated the role of frequency, pulse duration, total exposure time, and underlying mechanisms in order to unlock the full potential of PTHrP actions. Human MSC subjected to in vitro chondrogenesis for six weeks were exposed to 2.5 nM PTHrP(1‐34) pulses from days 7 to 42. Application frequency was increased from three times weekly (3 × 6 h/week) to daily maintaining either the duration of individual pulses (6 h/day) or total exposure time (18 h/week; 2.6 h/day). Daily PTHrP treatment significantly increased extracellular matrix deposition regardless of pulse duration and suppressed alkaline‐phosphatase activity by 87%. High total exposure time significantly reduced cell proliferation at day 14. Pulse duration was critically important to significantly reduce IHH expression, but irrelevant for PTHrP‐induced suppression of the hypertrophic markers MEF2C and IBSP. COL10A1, RUNX2, and MMP13 expression remained unaltered. Decreased IGFBP‐2, ‐3, and ‐6 expression suggested modulated IGF‐I availability in PTHrP groups, while drop of SOX9 protein levels during the PTHrP‐pulse may delay chondroblast formation and hypertrophy. Overall, the significantly optimized timing of PTHrP‐pulses demonstrated a vast potential to enhance chondrogenesis of MSC and suppress hypertrophy possibly via superior balancing of IGF‐ and SOX9‐related mechanisms. J. Cell. Physiol. 231: 2673–2681, 2016. © 2016 Wiley Periodicals, Inc. Daily PTHrP treatment significantly increased extracellular matrix deposition regardless of pulse duration and suppressed alkaline‐phosphatase activity by 87%. Pulse duration was critically important to significantly reduce IHH expression, but irrelevant for PTHrP‐induced suppression of the hypertrophic markers MEF2C and IBSP. Decreased IGFBP‐2 and ‐6 expression suggested modulated IGF‐I availability in PTHrP groups while drop of SOX9 protein levels during the PTHrP‐pulse may delay chondroblast formation and hypertrophy. Overall, the significantly optimized timing of PTHrP‐pulses demonstrated a vast potential to enhance chondrogenesis of MSC and suppress hypertrophy possibly via superior balancing of IGF‐ and SOX9‐related mechanisms.

  March 21, 2016   doi: 10.1002/jcp.25369   open full text
• A New In Vitro Co‐Culture Model Using Magnetic Force‐Based Nanotechnology.
  Hiroki Takanari, Keiko Miwa, XianMing Fu, Junichi Nakai, Akira Ito, Kousuke Ino, Hiroyuki Honda, Wataru Tonomura, Satoshi Konishi, Tobias Opthof, Marcel AG van der Heyden, Itsuo Kodama, Jong‐Kook Lee.
  Journal of Cellular Physiology. March 16, 2016
  Skeletal myoblast (SkMB) transplantation has been conducted as a therapeutic strategy for severe heart failure. However, arrhythmogenicity following transplantation remains unsolved. We developed an in vitro model of myoblast transplantation with “patterned” or “randomly‐mixed” co‐culture of SkMBs and cardiomyocytes enabling subsequent electrophysiological, and arrhythmogenic evaluation. SkMBs were magnetically labeled with magnetite nanoparticles and co‐cultured with neonatal rat ventricular myocytes (NRVMs) on multi‐electrode arrays. SkMBs were patterned by a magnet beneath the arrays. Excitation synchronicity was evaluated by Ca2+ imaging using a gene‐encoded Ca2+ indicator, G‐CaMP2. In the monoculture of NRVMs (control), conduction was well‐organized. In the randomly‐mixed co‐culture of NRVMs and SkMBs (random group), there was inhomogeneous conduction from multiple origins. In the “patterned” co‐culture where an en bloc SKMB‐layer was inserted into the NRVM‐layer, excitation homogenously propagated although conduction was distorted by the SkMB‐area. The 4‐mm distance conduction time (CT) in the random group was significantly longer (197 ± 126 ms) than in control (17 ± 3 ms). In the patterned group, CT through NRVM‐area did not change (25 ± 3 ms), although CT through the SkMB‐area was significantly longer (132 ± 77 ms). The intervals between spontaneous excitation varied beat‐to‐beat in the random group, while regular beating was recorded in the control and patterned groups. Synchronized Ca2+ transients of NRVMs were observed in the patterned group, whereas those in the random group were asynchronous. Patterned alignment of SkMBs is feasible with magnetic nanoparticles. Using the novel in vitro model mimicking cell transplantation, it may become possible to predict arrhythmogenicity due to heterogenous cell transplantation. J. Cell. Physiol. 231: 2249–2256, 2016. © 2016 Wiley Periodicals, Inc.

  March 16, 2016   doi: 10.1002/jcp.25342   open full text
• Alteration in Phospholipidome Profile of Myoblast H9c2 Cell Line in a Model of Myocardium Starvation and Ischemia.
  Bebiana Sousa, Tânia Melo, Ana Campos, Ana S.P. Moreira, Elisabete Maciel, Pedro Domingues, Rita Pereira Carvalho, Teresa Ribeiro Rodrigues, Henrique Girão, Maria Rosario M. Domingues.
  Journal of Cellular Physiology. March 16, 2016
  Myocardium infarction is one of the most deathly cardiovascular diseases. It is characterized by myocardium ischemia as a result of nutrients depletion and hypoxia. The cell can respond to this injury by autophagy or apoptosis, which determines the evolution and possible recovery of the myocardium infarction. Lipids play an important role in cardiovascular disease. However reports stating lipidome variations in cardiovascular disease are scarce and the role that lipids play in this pathological condition is not completely understood. The aim of this work was to identify changes in lipid profile of a myoblast H9c2 cell line under starvation and ischemia, to better understand and recognize new biomarkers for myocardial infarction. Lipidomic profile was evaluated by HILIC‐LC‐MS and GC‐MS. Cardiac cells showed alterations in phosphatidylcholines PC (34:1) and PC (36:2), lysophosphatidylcholines lyso PC(16:0), lysoPC(18:1) and lysoPC(18:0), phosphatidylethanolamine PE (34:1), phosphatidylserine PS (36:1), phosphatidylinositol PI (36:2), PI (38:3) and PI (38:5), sphingomyelin SM (34:1) and cardiolipins CL(68:4), CL(72:5) and CL(74:7) in ischemia and/or starvation, in comparison with control. Specific differences observed only in starvation were decrease of SM (34:1) and FA (20:4), and increase of PS (36:1). Differences observed only in ischemia were decrease of PC (36:2), lyso PC (16:0) and FA (18:1) and simultaneous increase of FA (16:0), and FA (18:0). Interestingly, PC (34:1) increased in ischemia and decreased in starvation. In conclusion, our work suggests that lipids are potential markers for evaluation of cell fate, either cell death or recovery, which will be useful to improve diagnosis and prognostic of cardiovascular diseases. J. Cell. Physiol. 231: 2266–2274, 2016. © 2016 Wiley Periodicals, Inc. This work identified changes in lipid profile of a myoblast H9c2 cell line under starvation and ischemia by HILIC‐LC‐MS and by GC‐MS. Specific differences of starvation were observed in SM, FA e PS, while in isquemia diferences were observed in PC and LysoPC and FA. Lipids are potential markers for evaluating cell's either cell death or recovery, which will be useful to improve diagnosis and prognostic of cardiovascular diseases.

  March 16, 2016   doi: 10.1002/jcp.25344   open full text
• Programmable Site‐Specific Nucleases for Targeted Genome Engineering in Higher Eukaryotes.
  Ganesan Govindan, Sivaprakash Ramalingam.
  Journal of Cellular Physiology. March 15, 2016
  Recent advances in the targeted genome engineering enable molecular biologists to generate sequence specific modifications with greater efficiency and higher specificity in complex eukaryotic genomes. Programmable site‐specific DNA cleavage reagents and cellular DNA repair mechanisms have made this possible. These reagents have become powerful tools for delivering a site‐specific genomic double‐strand break (DSB) at the desired chromosomal locus, which produces sequence alterations through error‐prone non‐homologous end joining (NHEJ) resulting in gene inactivations/knockouts. Alternatively, the DSB can be repaired through homology‐directed repair (HDR) using a donor DNA template, which leads to the introduction of desired sequence modifications at the predetermined site. Here, we summarize the role of three classes of nucleases; zinc finger nucleases (ZFNs), transcription activator like effector nucleases (TALENs), and clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) system in achieving targeted genome modifications. Further, we discuss the progress towards the applications of programmable site‐specific nucleases (SSNs) in treating human diseases and other biological applications in economically important higher eukaryotic organisms such as plants and livestock. J. Cell. Physiol. 231: 2380–2392, 2016. © 2016 Wiley Periodicals, Inc. In this review article, we discuss the properties of programmable site‐specific nucleases and their biomedical and biological applications in higher eukaryotic organisms.

  March 15, 2016   doi: 10.1002/jcp.25367   open full text
• Deregulated E2F5/p38/SMAD3 Circuitry Reinforces the Pro‐Tumorigenic Switch of TGFβ Signaling in Prostate Cancer.
  Subhadipa Majumder, Ankur Bhowal, Sanmitra Basu, Pritha Mukherjee, Urmi Chatterji, Sanghamitra Sengupta.
  Journal of Cellular Physiology. March 14, 2016
  Transforming growth factor‐β signaling exerts divergent effects on normal and cancer cells, although mechanism underlying this differential behavior remains unclear. In this study, expression of 94 genes pertaining to the TGF‐β signaling pathway was compared between tumor and benign tissue samples from the human prostate gland to identify major discriminators driving prostate carcinogenesis. E2F5 was identified as one of the most deregulated genes in prostate cancer tissues, predominantly in samples with Gleason‐score 6. Expression of other deregulated components of TGF‐β signaling was examined by qRT‐PCR, Western blot, and immune‐staining. Function of E2F5 and p38 in prostate cancer was investigated using siRNA‐treatment of PC3 cell‐line followed by analyses of associated components and cell cycle. Observations revealed that E2F5 overexpression was accompanied by significantly higher phosphorylation of SMAD3 at Ser‐208 in the linker region (pSMAD3L) and p38 in tumor tissue. A striking difference in SMAD3 phosphorylation, marked by preponderance of pSMAD3L and pSMAD3C (Ser‐423 and 425) in tumor and benign tissues, respectively was noted. Co‐localization of E2F5 with pSMAD3L in the nuclei of tumor and PC3 cells indicated a functional interface between the proteins. Downregulation of E2F5 and p38 in PC3 cells resulted in marked reduction of phosphorylation of SMAD3 and perturbation of cell cycle with an arrest of cells in G1. Our findings unearthed that E2F5/p38 axis played a cardinal role in uncontrolled cellular proliferation in prostate cancer through pSMAD3L activation. It also underscores a strong potential for E2F5 to be incorporated as a tool in early detection of prostate cancer. J. Cell. Physiol. 231: 2482–2492, 2016. © 2016 Wiley Periodicals, Inc. Transforming growth factor‐β signaling exerts divergent effects on normal and cancer cells. Our study demonstrates that dysregulated E2F5/p38/pSMAD3L axis plays an important role in converting tumor‐suppressive TGFβ signaling into pro‐tumorigenic signaling.

  March 14, 2016   doi: 10.1002/jcp.25361   open full text
• Radiation‐Induced RhoGDIβ Cleavage Leads to Perturbation of Cell Polarity: A Possible Link to Cancer Spreading.
  Mamoru Fujiwara, Mayumi Okamoto, Masato Hori, Hiroshi Suga, Hiroshi Jikihara, Yuka Sugihara, Fumio Shimamoto, Toshio Mori, Koichi Nakaoji, Kazuhiko Hamada, Takahide Ota, Ralf Wiedemuth, Achim Temme, Masaaki Tatsuka.
  Journal of Cellular Physiology. March 14, 2016
  The equilibrium between proliferation and apoptosis is tightly balanced to maintain tissue homeostasis in normal tissues and even in tumors. Achieving and maintaining such a balance is important for cancer regrowth and spreading after cytotoxic treatments. Caspase‐3 activation and tumor cell death following anticancer therapy as well as accompanying cell death pathways are well characterized, but their association to homeostasis of cancerous tissue and tumor progression remains poorly understood. Here we proposed a novel mechanism of cancer spreading induced by caspase‐3. RhoGDIβ, known as a direct cleavage substrate of caspase‐3, is overexpressed in many epithelial cancers. The N‐terminal‐truncated RhoGDIβ (ΔN‐RhoGDIβ) is accumulated in caspase‐3‐activated cells. Stable expression of ΔN‐RhoGDIβ in HeLa cells did not induce apoptosis, but impaired directional cell migration in a wound‐healing assay accompanied by a perturbed direction of cell division at the wound edge. Subcellular protein fractionation experiments revealed that ΔN‐RhoGDIβ but not wild‐type RhoGDIβ was present in the detergent‐soluble cytoplasmic and nuclear fractions and preferentially associated with Cdc42. Furthermore, Cdc42 activity was constitutively inhibited by stable expression of ΔN‐RhoGDIβ, resulting in increased radiation‐induced compensatory proliferation linking to RhoA activation. Thus, ΔN‐RhoGDIβ dominant‐negatively regulates Cdc42 activity and contributes to loss of polarity‐related functions. The caspase‐3‐cleaved RhoGDIβ is a possible determinant to promote cancer spreading due to deregulation of directional organization of tumor cell population and inhibition of default equilibrium between proliferation and apoptosis after cytotoxic damage. J. Cell. Physiol. 231: 2493–2505, 2016. © 2016 Wiley Periodicals, Inc. The N‐terminal‐truncated RhoGDIβ (ΔN‐RhoGDIβ) is accumulated in caspase‐3‐activated cells. ΔN‐RhoGDIβ dominant‐negatively regulates Cdc42 activity and contributes to loss of polarity‐related functions. ΔN‐RhoGDIβ is a possible determinant to promote cancer spreading due to deregulation of directional organization of tumor cell population and inhibition of default equilibrium between proliferation and apoptosis after cytotoxic damage.

  March 14, 2016   doi: 10.1002/jcp.25362   open full text
• The Androgen Receptor Regulates PPARγ Expression and Activity in Human Prostate Cancer Cells.
  Emuejevoke Olokpa, Adrienne Bolden, LaMonica V. Stewart.
  Journal of Cellular Physiology. March 14, 2016
  The peroxisome proliferator activated receptor gamma (PPARγ) is a ligand‐activated transcription factor that regulates growth and differentiation within normal prostate and prostate cancers. However the factors that control PPARγ within the prostate cancers have not been characterized. The goal of this study was to examine whether the androgen receptor (AR) regulates PPARγ expression and function within human prostate cancer cells. qRT‐PCR and Western blot analyses revealed nanomolar concentrations of the AR agonist dihydrotestosterone (DHT) decrease PPARγ mRNA and protein within the castration‐resistant, AR‐positive C4‐2 and VCaP human prostate cancer cell lines. The AR antagonists bicalutamide and enzalutamide blocked the ability of DHT to reduce PPARγ levels. In addition, siRNA mediated knockdown of AR increased PPARγ protein levels and ligand‐induced PPARγ transcriptional activity within the C4‐2 cell line. Furthermore, proteasome inhibitors that interfere with AR function increased the level of basal PPARγ and prevented the DHT‐mediated suppression of PPARγ. These data suggest that AR normally functions to suppress PPARγ expression within AR‐positive prostate cancer cells. To determine whether increases in AR protein would influence PPARγ expression and activity, we used lipofectamine‐based transfections to overexpress AR within the AR‐null PC‐3 cells. The addition of AR to PC‐3 cells did not significantly alter PPARγ protein levels. However, the ability of the PPARγ ligand rosiglitazone to induce activation of a PPARγ‐driven luciferase reporter and induce expression of FABP4 was suppressed in AR‐positive PC‐3 cells. Together, these data indicate AR serves as a key modulator of PPARγ expression and function within prostate tumors. J. Cell. Physiol. 231: 2664–2672, 2016. © 2016 Wiley Periodicals, Inc. The goal of this study was to determine whether the androgen receptor (AR) signaling pathway influences the expression and function of PPAR gamma within human prostate cancer cells. Our results demonstrate that the presence of AR suppresses PPAR gamma protein levels, and inhibits the ability of PPAR gamma agonists to regulate transcription and suppress prostate cancer cell proliferation.

  March 14, 2016   doi: 10.1002/jcp.25368   open full text
• miR‐19a, ‐19b, and ‐26b Mediate CTGF Expression and Pulmonary Fibroblast Differentiation.
  Yen‐Chou Chen, Bing‐Chang Chen, Chung‐Chi Yu, Shin‐Hua Lin, Chien‐Huang Lin.
  Journal of Cellular Physiology. March 14, 2016
  Although microRNA (miRNA) dysregulation with intracellular signaling cascade disruption has been demonstrated in the pathophysiology of pulmonary fibrosis, the relationship between miRNAs and intracellular signaling cascades in pulmonary fibrosis remains unclear. Using the human embryonic lung fibroblast cell line WI‐38, we observed endothelin‐1 (ET‐1)‐ and thrombin‐induced expression of the differentiation markers α‐smooth muscle actin (α‐SMA) and vimentin along with increased connective tissue growth factor (CTGF) protein expression. Decreased CTGF protein expression by CTGF siRNA significantly blocked ET‐1‐ and thrombin‐induced α‐SMA and vimentin expression in WI‐38 cells. Activation of the mitogen‐activated protein kinases (MAPKs) extracellular signal‐regulated kinase ERK, c‐Jun N‐terminal kinase (JNK), and p38 contributed to ET‐1‐ and thrombin‐induced CTGF, α‐SMA, and vimentin expression in WI‐38 cells. TargetScan Human, miRanda, and PicTar prediction algorithms were used to predict miRNAs with binding sites in the 3′ untranslated region (UTR) of CTGF mRNA. miR‐19a, ‐19b, and ‐26b were candidate miRNAs of CTGF. Direct binding of the candidate miRNAs to the 3′‐UTR of CTGF mRNA was verified through luciferase assay by using SV40‐promoter‐IRES‐driven luciferase containing the 3′‐UTR of CTGF mRNA as a reporter plasmid. ET‐1 and thrombin reduced candidate miRNA levels. Candidate miRNA overexpression significantly suppressed ET‐1‐ and thrombin‐induced CTGF expression and reduced α‐SMA and vimentin expression in the WI‐38 cells. Furthermore, candidate miRNA levels were decreased in the lung tissues of mice with bleomycin‐induced pulmonary fibrosis, and intratracheal application of miR‐19a, ‐19b, and 26b reduced the pulmonary fibrotic severity induced by bleomycin. This study is the first to demonstrate crosstalk between MAPK activation and reduction in miR‐19a, ‐19b, and ‐26b expression leading to lung fibroblast differentiation. J. Cell. Physiol. 231: 2236–2248, 2016. © 2016 Wiley Periodicals, Inc.

  March 14, 2016   doi: 10.1002/jcp.25341   open full text
• Characterization of Notch Signaling During Osteogenic Differentiation in Human Osteosarcoma Cell Line MG63.
  Alessia Ongaro, Agnese Pellati, Leila Bagheri, Paola Rizzo, Cristiana Caliceti, Leo Massari, Monica De Mattei.
  Journal of Cellular Physiology. March 11, 2016
  Osteogenic differentiation is a multi‐step process controlled by a complex molecular framework. Notch is an evolutionarily conserved intercellular signaling pathway playing a prominent role in cell fate and differentiation, although the mechanisms by which this pathway regulates osteogenesis remain controversial. This study aimed to investigate, in vitro, the involvement of Notch pathway during all the developmental stages of osteogenic differentiation in human osteosarcoma cell line MG63. Cells were cultured in basal condition (control) and in osteoinductive medium (OM). Notch inhibitors were also added in OM to block Notch pathway. During osteogenic differentiation, early (alkaline phosphatase activity and collagen type I) and late osteogenic markers (osteocalcin levels and matrix mineralization), as well as the gene expression of the main osteogenic transcription factors (Runx2, Osterix, and Dlx5) increased. Time dependent changes in the expression of specific Notch receptors were identified in OM versus control with a significant reduction in the expression of Notch1 and Notch3 receptors in the early phase of differentiation, and an increase of Notch2 and Notch4 receptors in the late phase. Among Notch nuclear target genes, Hey1 expression was significantly higher in OM than control, while Hes5 expression decreased. Osteogenic markers were reduced and Hey1 was significantly inhibited by Notch inhibitors, suggesting a role for Notch through the canonical pathway. In conclusion, Notch pathway might be involved with a dual role in osteogenesis of MG63, through the activation of Notch2, Notch4, and Hey1, inducing osteoblast differentiation and the depression of Notch1, Notch3, and Hes5, maintaining an undifferentiated status. J. Cell. Physiol. 231: 2652–2663, 2016. © 2016 Wiley Periodicals, Inc. This study investigated in vitro the involvement of Notch pathway during all the developmental stages of osteogenic differentiation in human osteosarcoma cell line MG63. During osteogenic differentiation, early and late osteogenic markers, as well as the gene expression of the main osteogenic transcription factors increased. Time dependent changes in the expression of specific Notch receptors and nuclear target genes were also identified. Our results showed that Notch pathway might be involved with a dual role in osteogenesis of MG63, through the activation of Notch2, Notch4, and Hey1, inducing osteoblast differentiation and the depression of Notch1, Notch3, and Hes5, maintaining an undifferentiated status.

  March 11, 2016   doi: 10.1002/jcp.25366   open full text
• Oncofetal Epigenetic Bivalency in Breast Cancer Cells: H3K4 and H3K27 Tri‐Methylation as a Biomarker for Phenotypic Plasticity.
  Terri L. Messier, Joseph R. Boyd, Jonathan A. R. Gordon, Janet L. Stein, Jane B. Lian, Gary S. Stein.
  Journal of Cellular Physiology. March 10, 2016
  Alterations in the epigenetic landscape are fundamental drivers of aberrant gene expression that contribute to cancer progression and pathology. Understanding specific modes of epigenetic regulation can be used to identify novel biomarkers or targets for therapeutic intervention to clinically treat solid tumors and leukemias. The bivalent marking of gene promoters by H3K4me3 and H3K27me3 is a primary mechanism to poise genes for expression in pluripotent embryonic stem cells (ESC). In this study we interrogated three well‐established mammary cell lines to model epigenetic programming observed among breast cancer subtypes. Evidence is provided for a distinct bivalent signature, activating and repressive histone marks co‐residing at the same gene promoter, in the MCF7 (ESR/PGR+) luminal breast cancer cell line. We identified a subset of genes, enriched for developmental pathways that regulate cellular phenotype and signaling, and partially recapitulate the bivalent character observed in ESC. We validated the biological relevance of this “oncofetal epigenetic” signature using data from ESR/PGR+ tumor samples from breast cancer patients. This signature of oncofetal epigenetic control is an informative biomarker and may provide novel therapeutic targets, selective for both recurring and treatment‐resistant cancers. J. Cell. Physiol. 231: 2474–2481, 2016. © 2016 Wiley Periodicals, Inc.

  March 10, 2016   doi: 10.1002/jcp.25359   open full text
• Current Controversies in the Management of Myeloma Bone Disease.
  Rebecca Silbermann, Garson David Roodman.
  Journal of Cellular Physiology. March 10, 2016
  Recent significant advances in the treatment of multiple myeloma have resulted in an improvement in median overall survival from 4.6 years, for patients diagnosed between 2001 and 2005, to 6.1 years, for those diagnosed between 2006 and 2010 (Kumar et al., 2014). However, myeloma bone lesions persist in the absence of active disease and continue to be frequent and significant causes of patient morbidity and contribute to mortality. While bisphosphonate therapy in combination with anti‐myeloma therapy remains the cornerstone of skeletal disease management in myeloma, open questions regarding the optimal management of patients with myeloma bone disease remain. This article will address when to initiate and stop bone‐targeted therapy in patients with monoclonal gammopathies, duration of bisphosphonate treatment in the era of more effective anti‐myeloma treatment, the role of bone resorption markers in determining the dosing schedule for anti‐resorptive therapy, risks and benefits of long term anti‐resorptive therapy, and whether anti‐resorptive therapies should be stopped to enhance the potential anabolic effects of proteasome antagonists and other anabolic agents. J. Cell. Physiol. 231: 2374–2379, 2016. © 2016 Wiley Periodicals, Inc.

  March 10, 2016   doi: 10.1002/jcp.25351   open full text
• Retracted: Identification of Novel Biomarkers for Pancreatic Cancer Using Integrated Transcriptomics With Functional Pathways Analysis.
  Xuan Zhang, Pan Tong, Jinyun Chen, Zenglin Pei, Xiaoyan Zhang, Weiping Chen, Jianqing Xu, Jin Wang.
  Journal of Cellular Physiology. March 10, 2016
  Retraction: ‘Identification of Novel Biomarkers for Pancreatic Cancer Using Integrated Transcriptomics With Functional Pathways Analysis’ by Zhang, X., Tong, P., Chen, J., Pei, Z., Zhang, X., Chen, W., Xu, J. and Wang, J. The above article from the Journal of Cellular Physiology, published online on 10 March 2016 in Wiley Online Library as Early View (http://onlinelibrary.wiley.com/enhanced/doi/10.1002/jcp.25353/), has been retracted by agreement between Gary Stein, the journal's Editor‐in‐Chief, and Wiley Periodicals, Inc. The retraction has been agreed following an investigation at the University of Texas, MD Anderson Cancer Center, which confirmed that the article was submitted and approved for publication by Dr. Jin Wang without acknowledgement of NIH funding received or the consent and authorship of Dr. Ann Killary and Dr. Subrata Sen, with whom the manuscript was originally drafted.

  March 10, 2016   doi: 10.1002/jcp.25353   open full text
• Phosphorylation Regulates Functions of ZEB1 Transcription Factor.
  M. Candelaria Llorens, Guadalupe Lorenzatti, Natalia L. Cavallo, Maria V. Vaglienti, Ana P. Perrone, Anne L. Carenbauer, Douglas S. Darling, Ana M. Cabanillas.
  Journal of Cellular Physiology. March 10, 2016
  ZEB1 transcription factor is important in both development and disease, including many TGFβ‐induced responses, and the epithelial‐to‐mesenchymal transition (EMT) by which many tumors undergo metastasis. ZEB1 is differentially phosphorylated in different cell types; however the role of phosphorylation in ZEB1 activity is unknown. Luciferase reporter studies and electrophoresis mobility shift assays (EMSA) show that a decrease in phosphorylation of ZEB1 increases both DNA‐binding and transcriptional repression of ZEB1 target genes. Functional analysis of ZEB1 phosphorylation site mutants near the second zinc finger domain (termed ZD2) show that increased phosphorylation (due to either PMA plus ionomycin, or IGF‐1) can inhibit transcriptional repression by either a ZEB1‐ZD2 domain clone, or full‐length ZEB1. This approach identifies phosphosites that have a substantial effect regulating the transcriptional and DNA‐binding activity of ZEB1. Immunoprecipitation with anti‐ZEB1 antibodies followed by western analysis with a phospho‐Threonine‐Proline‐specific antibody indicates that the ERK consensus site at Thr‐867 is phosphorylated in ZEB1. In addition to disrupting in vitro DNA‐binding measured by EMSA, IGF‐1‐induced MEK/ERK phosphorylation is sufficient to disrupt nuclear localization of GFP‐ZEB1 fusion clones. These data suggest that phosphorylation of ZEB1 integrates TGFβ signaling with other signaling pathways such as IGF‐1. J. Cell. Physiol. 231: 2205–2217, 2016. © 2016 Wiley Periodicals, Inc.

  March 10, 2016   doi: 10.1002/jcp.25338   open full text
• Lack of Dystrophin Affects Bronchial Epithelium in mdx Mice.
  Giuseppe Morici, Francesca Rappa, Francesco Cappello, Elisabetta Pace, Andrea Pace, Giuseppa Mudò, Grazia Crescimanno, Natale Belluardo, Maria R. Bonsignore.
  Journal of Cellular Physiology. March 10, 2016
  Mild exercise training may positively affect the course of Duchenne Muscular Dystrophy (DMD). Training causes mild bronchial epithelial injury in both humans and mice, but no study assessed the effects of exercise in mdx mice, a well known model of DMD. The airway epithelium was examined in mdx (C57BL/10ScSn‐Dmdmdx) mice, and in wild type (WT, C57BL/10ScSc) mice either under sedentary conditions (mdx‐SD, WT‐SD) or during mild exercise training (mdx‐EX, WT‐EX). At baseline, and after 30 and 45 days of training (5 d/wk for 6 weeks), epithelial morphology and markers of regeneration, apoptosis, and cellular stress were assessed. The number of goblet cells in bronchial epithelium was much lower in mdx than in WT mice under all conditions. At 30 days, epithelial regeneration (PCNA positive cells) was higher in EX than SD animals in both groups; however, at 45 days, epithelial regeneration decreased in mdx mice irrespective of training, and the percentage of apoptotic (TUNEL positive) cells was higher in mdx‐EX than in WT‐EX mice. Epithelial expression of HSP60 (marker of stress) progressively decreased, and inversely correlated with epithelial apoptosis (r = −0.66, P = 0.01) only in mdx mice. Lack of dystrophin in mdx mice appears associated with defective epithelial differentiation, and transient epithelial regeneration during mild exercise training. Hence, lack of dystrophin might impair repair in bronchial epithelium, with potential clinical consequences in DMD patients. J. Cell. Physiol. 231: 2218–2223, 2016. © 2016 Wiley Periodicals, Inc. Airway epithelium was studied in mdx and wild‐type mice at rest and during exercise training. The major finding of the study was that mdx mice showed very few goblet cells at all times, irrespective of training. In addition, epithelial regeneration sharply decreased, and apoptosis increased, late in the course of the study only in mdx mice, together with decreased expression of HSP60.

  March 10, 2016   doi: 10.1002/jcp.25339   open full text
• Skin Metabolite, Farnesyl Pyrophosphate, Regulates Epidermal Response to Inflammation, Oxidative Stress, and Migration.
  Irena Pastar, Olivera Stojadinovic, Andrew P. Sawaya, Rivka C. Stone, Linsey E. Lindley, Nkemcho Ojeh, Sasa Vukelic, Herbert H. Samuels, Marjana Tomic‐Canic.
  Journal of Cellular Physiology. March 09, 2016
  Skin produces cholesterol and a wide array of sterols and non‐sterol mevalonate metabolites, including isoprenoid derivative farnesyl pyrophosphate (FPP). To characterize FPP action in epidermis, we generated transcriptional profiles of primary human keratinocytes treated with zaragozic acid (ZGA), a squalene synthase inhibitor that blocks conversion of FPP to squalene resulting in endogenous accumulation of FPP. The elevated levels of intracellular FPP resulted in regulation of epidermal differentiation and adherens junction signaling, insulin growth factor (IGF) signaling, oxidative stress response and interferon (IFN) signaling. Immunosuppressive properties of FPP were evidenced by STAT‐1 downregulation and prominent suppression of its nuclear translocation by IFNγ. Furthermore, FPP profoundly downregulated genes involved in epidermal differentiation of keratinocytes in vitro and in human skin ex vivo. Elevated levels of FPP resulted in induction of cytoprotective transcriptional factor Nrf2 and its target genes. We have previously shown that FPP functions as ligand for the glucocorticoid receptor (GR), one of the major regulator of epidermal homeostasis. Comparative microarray analyses show significant but not complete overlap between FPP and glucocorticoid regulated genes, suggesting that FPP may have wider transcriptional impact. This was further supported by co‐transfection and chromatin immunoprecipitation experiments where we show that upon binding to GR, FPP recruits β‐catenin and, unlike glucocorticoids, recruits co‐repressor GRIP1 to suppress keratin 6 gene. These findings have many clinical implications related to epidermal lipid metabolism, response to glucocorticoid therapy as well as pleiotropic effects of cholesterol lowering therapeutics, statins. J. Cell. Physiol. 231: 2452–2463, 2016. © 2016 Wiley Periodicals, Inc.

  March 09, 2016   doi: 10.1002/jcp.25357   open full text
• The Cholinergic Signaling Responsible for the Expression of a Memory‐Related Protein in Primary Rat Cortical Neurons.
  Tsan‐Ju Chen, Shun‐Sheng Chen, Dean‐Chuan Wang, Hui‐Shan Hung.
  Journal of Cellular Physiology. March 08, 2016
  Cholinergic dysfunction in the brain is closely related to cognitive impairment including memory loss. In addition to the degeneration of basal forebrain cholinergic neurons, deficits in the cholinergic receptor signaling may also play an important role. In the present study, to examine the cholinergic signaling pathways responsible for the induction of a memory‐related postsynaptic protein, a cholinergic agonist carbachol was used to induce the expression of activity‐regulated cytoskeleton associated protein (Arc) in primary rat cortical neurons. After pretreating neurons with various antagonists or inhibitors, the levels of carbachol‐induced Arc protein expression were detected by Western blot analysis. The results show that carbachol induces Arc protein expression mainly through activating M1 acetylcholine receptors and the downstream phospholipase C pathway, which may lead to the activation of the MAPK/ERK signaling pathway. Importantly, carbachol‐mediated M2 receptor activation exerts negative effects on Arc protein expression and thus counteracts the enhanced effects of M1 activation. Furthermore, it is suggested for the first time that M1‐mediated enhancement of N‐methyl‐D‐aspartate receptor (NMDAR) responses, leading to Ca2+ entry through NMDARs, contributes to carbachol‐induced Arc protein expression. These findings reveal a more complete cholinergic signaling that is responsible for carbachol‐induced Arc protein expression, and thus provide more information for developing treatments that can modulate cholinergic signaling and consequently alleviate cognitive impairment. J. Cell. Physiol. 231: 2428–2438, 2016. © 2016 Wiley Periodicals, Inc. Cholinergic agonist carbachol induces the expression of a memory‐related protein, Arc, mainly through activating M1 acetylcholine receptors and the downstream phospholipase C pathway, leading to the activation of the MAPK/ERK signaling pathway. In addition, M1‐mediated enhancement of N‐methyl‐D‐aspartate receptor (NMDAR) responses, leading to Ca2+ entry through NMDARs, contributes to carbachol‐induced Arc protein expression.

  March 08, 2016   doi: 10.1002/jcp.25347   open full text
• Annexin A1 Is a Physiological Modulator of Neutrophil Maturation and Recirculation Acting on the CXCR4/CXCL12 Pathway.
  Isabel Daufenback Machado, Marina Spatti, Araceli Hastreiter, José Roberto Santin, Ricardo Ambrósio Fock, Cristiane Damas Gil, Sonia Maria Oliani, Mauro Perretti, Sandra Helena Poliselli Farsky.
  Journal of Cellular Physiology. March 08, 2016
  Neutrophil production and traffic in the body compartments is finely controlled, and the strong evidences support the role of CXCL12/CXCR4 pathway on neutrophil trafficking to and from the bone marrow (BM). We recently showed that the glucocorticoid‐regulated protein, Annexin A1 (AnxA1) modulates neutrophil homeostasis and here we address the effects of AnxA1 on steady‐state neutrophil maturation and trafficking. For this purpose, AnxA1−/− and Balb/C wild‐type mice (WT) were donors of BM granulocytes and mesenchymal stem cells and blood neutrophils. In vivo treatments with the pharmacological AnxA1 mimetic peptide (Ac2‐26) or the formyl peptide receptor (FPR) antagonist (Boc‐2) were used to elucidate the pathway of AnxA1 action, and with the cytosolic glucocorticoid antagonist receptor RU 38486. Accelerated maturation of BM granulocytes was detected in AnxA1−/− and Boc2‐treated WT mice, and was reversed by treatment with Ac2‐26 in AnxA1−/− mice. AnxA1 and FPR2 were constitutively expressed in bone marrow granulocytes, and their expressions were reduced by treatment with RU38486. Higher numbers of CXCR4+ neutrophils were detected in the circulation of AnxA1−/− or Boc2‐treated WT mice, and values were rescued in Ac2‐26‐treated AnxA1−/− mice. Although circulating neutrophils of AnxA1−/− animals were CXCR4+, they presented reduced CXCL12‐induced chemotaxis. Moreover, levels of CXCL12 were reduced in the bone marrow perfusate and in the mesenchymal stem cell supernatant from AnxA1−/− mice, and in vivo and in vitro CXCL12 expression was re‐established after Ac2‐26 treatment. Collectively, these data highlight AnxA1 as a novel determinant of neutrophil maturation and the mechanisms behind blood neutrophil homing to BM via the CXCL12/CXCR4 pathway. J. Cell. Physiol. 231: 2418–2427, 2016. © 2016 Wiley Periodicals, Inc.

  March 08, 2016   doi: 10.1002/jcp.25346   open full text
• Involvement of Transient Receptor Potential Cation Channel Vanilloid 1 (TRPV1) in Myoblast Fusion.
  Mitsutoshi Kurosaka, Yuji Ogura, Toshiya Funabashi, Tatsuo Akema.
  Journal of Cellular Physiology. March 08, 2016
  The mechanisms that underlie the complex process of muscle regeneration after injury remain unknown. Transient receptor potential cation channel vanilloid 1 (TRPV1) is expressed in several cell types, including skeletal muscle, and is activated by high temperature and by certain molecules secreted during tissue inflammation. Severe inflammation and local temperature perturbations are induced during muscle regeneration, which suggests that TRPV1 might be activated and involved in the process. The aim of this study, was to clarify the role of TRPV1 in the myogenic potential of satellite cells responsible for muscle regeneration. We found that mRNA and protein levels of TRPV1 increased during regeneration after cardiotoxin (CTX)‐induced muscle injury in mice. Using isolated mouse satellite cells (i.e., myoblasts), we observed that activation of TRPV1 by its agonist capsaicin (CAP) augmented myogenin protein levels. Whereas CAP did not alter myoblast proliferation, it facilitated myoblast fusion (evaluated using myonucleii number per myotube and fusion index). In contrast, suppression of TRPV1 by siRNA impaired myoblast fusion. Using mice, we also demonstrated that intramuscular injection of CAP facilitated muscle repair after CTX‐induced muscle injury. Moreover, we showed that these roles of TRPV1 might be mediated by interleukin‐4 and calcium signaling during myoblast fusion. Collectively, these results suggest that TRPV1 underlies normal myogenesis through promotion of myoblast fusion. J. Cell. Physiol. 231: 2275–2285, 2016. © 2016 Wiley Periodicals, Inc.

  March 08, 2016   doi: 10.1002/jcp.25345   open full text
• Differential Response of Human Hepatocyte Chromatin to HDAC Inhibitors as a Function of Microenvironmental Glucose Level.
  Marina Barreto Felisbino, Thiago Alves da Costa, Maria Silvia Viccari Gatti, Maria Luiza Silveira Mello.
  Journal of Cellular Physiology. March 06, 2016
  Diabetes is a complex multifactorial disorder characterized by chronic hyperglycemia due to impaired insulin secretion. Recent observations suggest that the complexity of the disease cannot be entirely accounted for genetic predisposition and a compelling argument for an epigenetic component is rapidly emerging. The use of histone deacetylase inhibitor (HDACi) in clinical setting is an emerging area of investigation. In this study, we have aimed to understand and compare the response of hepatocyte chromatin to valproic acid (VPA) and trichostatin A (TSA) treatments under normoglycemic or hyperglycemic conditions to expand our knowledge about the consequences of HDACi treatment in a diabetes cell model. Under normoglycemic conditions, these treatments promoted chromatin remodeling, as assessed by image analysis and H3K9ac and H3K9me2 abundance. Simultaneously, H3K9ac marks shifted to the nuclear periphery accompanied by HP1 dissociation from the heterochromatin and a G1 cell cycle arrest. More striking changes in the cell cycle progression and mitotic ratios required drastic treatment. Under hyperglycemic conditions, high glucose per se promoted chromatin changes similar to those promoted by VPA and TSA. Nonetheless, these results were not intensified in cells treated with HDACis under hyperglycemic conditions. Despite the absence of morphological changes being promoted, HDACi treatment seems to confer a physiological meaning, ameliorating the cellular hyperglycemic state through reduction of glucose production. These observations allow us to conclude that the glucose level to which the hepatocytes are subjected affects how chromatin responds to HDACi and their action under high‐glucose environment might not reflect on chromatin remodeling. J. Cell. Physiol. 231: 2257–2265, 2016. © 2016 Wiley Periodicals, Inc. In this study, we have aimed to understand and compare the response of hepatocyte chromatin to valproic acid (VPA) and trichostatin A (TSA) treatments under normoglycemic or hyperglycemic conditions to expand our knowledge about the consequences of histone deacetylase inhibitors (HDACi) treatment in a diabetes cell model. Under normoglycemic conditions, these treatments promoted chromatin remodeling, as assessed by image analysis and H3K9ac and H3K9me2 abundance. Simultaneously, H3K9ac marks shifted to the nuclear periphery accompanied by HP1 dissociation from the heterochromatin and a G1 cell cycle arrest. Under hyperglycemic conditions, high glucose per se promoted chromatin changes similar to those promoted by VPA and TSA. Nonetheless, these results were not intensified in cells treated with HDACis under hyperglycemic conditions. Despite absence of morphological changes being promoted, HDACi treatment seems to confer a physiological meaning, ameliorating the cellular hyperglycemic state through reduction of glucose production.

  March 06, 2016   doi: 10.1002/jcp.25343   open full text
• Absence of K‐Ras Reduces Proliferation and Migration But Increases Extracellular Matrix Synthesis in Fibroblasts.
  José M. Muñoz‐Félix, Isabel Fuentes‐Calvo, Cristina Cuesta, Nélida Eleno, Piero Crespo, José M. López‐Novoa, Carlos Martínez‐Salgado.
  Journal of Cellular Physiology. March 06, 2016
  The involvement of Ras‐GTPases in the development of renal fibrosis has been addressed in the last decade. We have previously shown that H‐ and N‐Ras isoforms participate in the regulation of fibrosis. Herein, we assessed the role of K‐Ras in cellular processes involved in the development of fibrosis: proliferation, migration, and extracellular matrix (ECM) proteins synthesis. K‐Ras knockout (KO) mouse embryonic fibroblasts (K‐ras−/−) stimulated with transforming growth factor‐β1 (TGF‐β1) exhibited reduced proliferation and impaired mobility than wild‐type fibroblasts. Moreover, an increase on ECM production was observed in K‐Ras KO fibroblasts in basal conditions. The absence of K‐Ras was accompanied by reduced Ras activation and ERK phosphorylation, and increased AKT phosphorylation, but no differences were observed in TGF‐β1‐induced Smad signaling. The MEK inhibitor U0126 decreased cell proliferation independently of the presence of K‐ras but reduced migration and ECM proteins expression only in wild‐type fibroblasts, while the PI3K‐AKT inhibitor LY294002 decreased cell proliferation, migration, and ECM synthesis in both types of fibroblasts. Thus, our data unveil that K‐Ras and its downstream effector pathways distinctively regulate key biological processes in the development of fibrosis. Moreover, we show that K‐Ras may be a crucial mediator in TGF‐β1‐mediated effects in this cell type. J. Cell. Physiol. 231: 2224–2235, 2016. © 2016 Wiley Periodicals, Inc.

  March 06, 2016   doi: 10.1002/jcp.25340   open full text
• Dynamic Histone Acetylation of H3K4me3 Nucleosome Regulates MCL1 Pre‐mRNA Splicing.
  Dilshad H. Khan, Carolina Gonzalez, Nikesh Tailor, Mohammad K. Hamedani, Etienne Leygue, James R. Davie.
  Journal of Cellular Physiology. March 06, 2016
  Pre‐mRNA splicing is a cotranscriptional process affected by the chromatin architecture along the body of coding genes. Recruited to the pre‐mRNA by splicing factors, histone deacetylases (HDACs) and K‐acetyltransferases (KATs) catalyze dynamic histone acetylation along the gene. In colon carcinoma HCT 116 cells, HDAC inhibition specifically increased KAT2B occupancy as well as H3 and H4 acetylation of the H3K4 trimethylated (H3K4me3) nucleosome positioned over alternative exon 2 of the MCL1 gene, an event paralleled with the exclusion of exon 2. These results were reproduced in MDA‐MB‐231, but not in MCF7 breast adenocarcinoma cells. These later cells have much higher levels of demethylase KDM5B than either HCT 116 or MDA‐MB‐231 cells. We show that H3K4me3 steady‐state levels and H3K4me3 occupancy at the end of exon 1 and over exon 2 of the MCL1 gene were lower in MCF7 than in MDA‐MB‐231 cells. Furthermore, in MCF7 cells, there was minimal effect of HDAC inhibition on H3/H4 acetylation and H3K4me3 levels along the MCL1 gene and no change in pre‐mRNA splicing choice. These results show that, upon HDAC inhibition, the H3K4me3 mark plays a critical role in the exclusion of exon 2 from the MCL1 pre‐mRNA. J. Cell. Physiol. 231: 2196–2204, 2016. © 2016 Wiley Periodicals, Inc.

  March 06, 2016   doi: 10.1002/jcp.25337   open full text
• Nitric Oxide Chemical Donor Affects the Early Phases of In Vitro Wound Healing Process.
  Cristina La Torre, Benedetta Cinque, Francesca Lombardi, Gianfranca Miconi, Paola Palumbo, Zoran Evtoski, Giuseppe Placidi, Donatella Fanini, Anna Maria Cimini, Elisabetta Benedetti, Maurizio Giuliani, Maria Grazia Cifone.
  Journal of Cellular Physiology. March 06, 2016
  An artificial wound in a confluent monolayer of human keratinocyte HaCaT cells or mouse embryo fibroblast Swiss NIH 3T3 cells was used to analyze the effects of the nitric oxide (NO) chemical donor, S‐nitroso‐N‐acetylpenicillamine (SNAP). SNAP exposure promoted an enhanced rate of wound closure and accelerated motility of both keratinocytes and fibroblasts compared to control cells. The wounded monolayer cultures of HaCaT and NIH 3T3 cells, treated with or without SNAP, were monitored under a phase contrast microscope. Structural and ultrastructural modifications were analyzed by scanning electron microscopy (SEM). The images were captured by a digital camera at different time points (0–28 h) and the wound area was analyzed through software included in Matlab®. As early as 15 min, SNAP induced significant cytoskeletal remodeling, as shown by immunostaining (phalloidin‐labelling), which in turn was associated with increased filopodium number and length rise. NO donor treatment also induced overexpression of Ki‐67 protein, a typical marker of cell proliferation, as shown by immunostaining. Both SNAP‐induced migration and proliferation were antagonized by the NO‐sensitive GC inhibitor 1H‐[1,2,4]oxadiazolo[‐4,3‐a]quinoxalin‐1‐one (ODQ), which suggests activation of the NO/cGMP signalling cascade in the observed SNAP‐induced effects in the early stages of the healing process. Moreover, we provide evidence that PPAR‐β antagonist (GSK0660) may interfere with NO‐mediated wound healing process. J. Cell. Physiol. 231: 2185–2195, 2016. © 2016 Wiley Periodicals, Inc.

  March 06, 2016   doi: 10.1002/jcp.25331   open full text
• First Evidence of DAAM1 Localization During the Post‐Natal Development of Rat Testis and in Mammalian Sperm.
  Paolo Pariante, Raffaele Dotolo, Massimo Venditti, Diana Ferrara, Aldo Donizetti, Francesco Aniello, Sergio Minucci.
  Journal of Cellular Physiology. March 06, 2016
  Dishevelled‐associated activator of morphogenesis 1 (DAAM1) is a formin‐family protein involved in nucleation of unbranched actin filaments and in cytoskeletal organization through Wnt‐Dishevelled PCP pathway, which participates in essential biological processes, such as cell polarity, movement, and adhesion during morphogenesis and organogenesis. While its role has been investigated during development and in somatic cells, its potential association with the germinal compartment and reproduction is still unexplored. In this work, we assessed the possible association of DAAM1 with the morphogenesis of rat testis. We studied its expression and profiled its localization versus actin and tubulin, during the first wave of spermatogenesis and in the adult gonad (from 7 to 60 dpp). We show that, in mitotic phases, DAAM1 shares its localization with actin in Sertoli cells, gonocytes, and spermatogonia. Later, during meiosis, both proteins are found in spermatocytes, while only actin is detectable at the forming blood‐testis barrier. DAAM1, then, follows the development of the acrosome system throughout spermiogenesis, and it is finally retained inside the cytoplasmic droplet in mature gametes, as corroborated by additional immunolocalization data on both rat and human sperm. Unlike the DAAM1, actin keeps its localization in Sertoli cells, and tubulin is associated with their protruding cytoplasm during the process. Our data support, for the first time, the hypothesis of a role for DAAM1 in cytoskeletal organization during Mammalian testis morphogenesis and gamete progression, while also hinting at its possible investigation as a morphological marker of germ cell and sperm physiology. J. Cell. Physiol. 231: 2172–2184, 2016. © 2016 Wiley Periodicals, Inc.

  March 06, 2016   doi: 10.1002/jcp.25330   open full text
• Emerging From the Unknown: Structural and Functional Features of Agnoprotein of Polyomaviruses.
  A. Sami Saribas, Pascale Coric, Anahit Hamazaspyan, William Davis, Rachael Axman, Martyn K. White, Magid Abou‐Gharbia, Wayne Childers, Jon H. Condra, Serge Bouaziz, Mahmut Safak.
  Journal of Cellular Physiology. February 24, 2016
  Agnoprotein is an important regulatory protein of polyomaviruses, including JCV, BKV, and SV40. In the absence of its expression, these viruses are unable to sustain their productive life cycle. It is a highly basic phosphoprotein that localizes mostly to the perinuclear area of infected cells, although a small amount of the protein is also found in nucleus. Much has been learned about the structure and function of this important regulatory protein in recent years. It forms highly stable dimers/oligomers in vitro and in vivo through its Leu/Ile/Phe‐rich domain. Structural NMR studies revealed that this domain adopts an alpha‐helix conformation and plays a critical role in the stability of the protein. It associates with cellular proteins, including YB‐1, p53, Ku70, FEZ1, HP1α, PP2A, AP‐3, PCNA, and α‐SNAP; and viral proteins, including small t antigen, large T antigen, HIV‐1 Tat, and JCV VP1; and significantly contributes the viral transcription and replication. This review summarizes the recent advances in the structural and functional properties of this important regulatory protein. J. Cell. Physiol. 231: 2115–2127, 2016. © 2016 Wiley Periodicals, Inc.

  February 24, 2016   doi: 10.1002/jcp.25329   open full text
• Corticosteroid‐Induced MKP‐1 Represses Pro‐Inflammatory Cytokine Secretion by Enhancing Activity of Tristetraprolin (TTP) in ASM Cells.
  Pavan Prabhala, Kristin Bunge, Qi Ge, Alaina J. Ammit.
  Journal of Cellular Physiology. February 16, 2016
  Exaggerated cytokine secretion drives pathogenesis of a number of chronic inflammatory diseases, including asthma. Anti‐inflammatory pharmacotherapies, including corticosteroids, are front‐line therapies and although they have proven clinical utility, the molecular mechanisms responsible for their actions are not fully understood. The corticosteroid‐inducible gene, mitogen‐activated protein kinase (MAPK) phosphatase 1 (MKP‐1, DUSP1) has emerged as a key molecule responsible for the repressive effects of steroids. MKP‐1 is known to deactivate p38 MAPK phosphorylation and can control the expression and activity of the mRNA destabilizing protein—tristetraprolin (TTP). But whether corticosteroid‐induced MKP‐1 acts via p38 MAPK‐mediated modulation of TTP function in a pivotal airway cell type, airway smooth muscle (ASM), was unknown. While pretreatment of ASM cells with the corticosteroid dexamethasone (preventative protocol) is known to reduce ASM synthetic function in vitro, the impact of adding dexamethasone after stimulation (therapeutic protocol) had not been explored. Whether dexamethasone modulates TTP in a p38 MAPK‐dependent manner in this cell type was also unknown. We address this herein and utilize an in vitro model of asthmatic inflammation where ASM cells were stimulated with the pro‐asthmatic cytokine tumor necrosis factor (TNF) and the impact of adding dexamethasone 1 h after stimulation assessed. IL‐6 mRNA expression and protein secretion was significantly repressed by dexamethasone acting in a temporally distinct manner to increase MKP‐1, deactivate p38 MAPK, and modulate TTP phosphorylation status. In this way, dexamethasone‐induced MKP‐1 acts via p38 MAPK to switch on the mRNA destabilizing function of TTP to repress pro‐inflammatory cytokine secretion from ASM cells. J. Cell. Physiol. 231: 2153–2158, 2016. © 2016 Wiley Periodicals, Inc.

  February 16, 2016   doi: 10.1002/jcp.25327   open full text
• Fatty Acids and Breast Cancer: Make Them on Site or Have Them Delivered.
  William B. Kinlaw, Paul W. Baures, Leslie E. Lupien, Wilson L. Davis, Nancy B. Kuemmerle.
  Journal of Cellular Physiology. February 16, 2016
  Brisk fatty acid (FA) production by cancer cells is accommodated by the Warburg effect. Most breast and other cancer cell types are addicted to fatty acids (FA), which they require for membrane phospholipid synthesis, signaling purposes, and energy production. Expression of the enzymes required for FA synthesis is closely linked to each of the major classes of signaling molecules that stimulate BC cell proliferation. This review focuses on the regulation of FA synthesis in BC cells, and the impact of FA, or the lack thereof, on the tumor cell phenotype. Given growing awareness of the impact of dietary fat and obesity on BC biology, we will also examine the less‐frequently considered notion that, in addition to de novo FA synthesis, the lipolytic uptake of preformed FA may also be an important mechanism of lipid acquisition. Indeed, it appears that cancer cells may exist at different points along a “lipogenic‐lipolytic axis,” and FA uptake could thwart attempts to exploit the strict requirement for FA focused solely on inhibition of de novo FA synthesis. Strategies for clinically targeting FA metabolism will be discussed, and the current status of the medicinal chemistry in this area will be assessed. J. Cell. Physiol. 231: 2128–2141, 2016. © 2016 Wiley Periodicals, Inc.

  February 16, 2016   doi: 10.1002/jcp.25332   open full text
• Zebrafish as a Model for the Study of Chaperonopathies.
  Gianfranco Bellipanni, Francesco Cappello, Federica Scalia, Everly Conway de Macario, Alberto J.L. Macario, Antonio Giordano.
  Journal of Cellular Physiology. February 16, 2016
  There is considerable information on the clinical manifestations and mode of inheritance for many genetic chaperonopathies but little is known on the molecular mechanisms underlying the cell and tissue abnormalities that characterize them. This scarcity of knowledge is mostly due to the lack of appropriate animal models that mimic closely the human molecular, cellular, and histological characteristics. In this article we introduce zebrafish as a suitable model to study molecular and cellular mechanisms pertaining to human chaperonopathies. Genetic chaperonopathies manifest themselves from very early in life so it is necessary to examine the impact of mutant chaperone genes during development, starting with fertilization and proceeding throughout the entire ontogenetic process. Zebrafish is amenable to such developmental analysis as well as studies during adulthood. In addition, the zebrafish genome contains a wide range of genes encoding proteins similar to those that form the chaperoning system of humans. This, together with the availability of techniques for genetic manipulations and for examination of all stages of development, makes zebrafish the organism of choice for the analysis of the molecular features and pathogenic mechanisms pertaining to human chaperonopathies. J. Cell. Physiol. 231: 2107–2114, 2016. © 2016 Wiley Periodicals, Inc.

  February 16, 2016   doi: 10.1002/jcp.25319   open full text
• Bee Venom Accelerates Wound Healing in Diabetic Mice by Suppressing Activating Transcription Factor‐3 (ATF‐3) and Inducible Nitric Oxide Synthase (iNOS)‐Mediated Oxidative Stress and Recruiting Bone Marrow‐Derived Endothelial Progenitor Cells.
  Gamal Badr, Wael N. Hozzein, Badr M. Badr, Ahmad Al Ghamdi, Heba M. Saad Eldien, Olivier Garraud.
  Journal of Cellular Physiology. February 15, 2016
  Multiple mechanisms contribute to impaired diabetic wound healing including impaired neovascularization and deficient endothelial progenitor cell (EPC) recruitment. Bee venom (BV) has been used as an anti‐inflammatory agent for the treatment of several diseases. Nevertheless, the effect of BV on the healing of diabetic wounds has not been studied. Therefore, in this study, we investigated the impact of BV on diabetic wound closure in a type I diabetic mouse model. Three experimental groups were used: group 1, non‐diabetic control mice; group 2, diabetic mice; and group 3, diabetic mice treated with BV. We found that the diabetic mice exhibited delayed wound closure characterized by a significant decrease in collagen production and prolonged elevation of inflammatory cytokines levels in wounded tissue compared to control non‐diabetic mice. Additionally, wounded tissue in diabetic mice revealed aberrantly up‐regulated expression of ATF‐3 and iNOS followed by a marked elevation in free radical levels. Impaired diabetic wound healing was also characterized by a significant elevation in caspase‐3, ‐8, and ‐9 activity and a marked reduction in the expression of TGF‐β and VEGF, which led to decreased neovascularization and angiogenesis of the injured tissue by impairing EPC mobilization. Interestingly, BV treatment significantly enhanced wound closure in diabetic mice by increasing collagen production and restoring the levels of inflammatory cytokines, free radical, TGF‐β, and VEGF. Most importantly, BV‐treated diabetic mice exhibited mobilized long‐lived EPCs by inhibiting caspase activity in the wounded tissue. Our findings reveal the molecular mechanisms underlying improved diabetic wound healing and closure following BV treatment. J. Cell. Physiol. 231: 2159–2171, 2016. © 2016 Wiley Periodicals, Inc.

  February 15, 2016   doi: 10.1002/jcp.25328   open full text
• Polyglucosan Molecules Induce Mitochondrial Impairment and Apoptosis in Germ Cells Without Affecting the Integrity and Functionality of Sertoli Cells.
  Franz Villarroel‐Espíndola, Cynthia Tapia, Roxana González‐Stegmaier, Ilona I. Concha, Juan Carlos Slebe.
  Journal of Cellular Physiology. February 11, 2016
  Glycogen is the main storage form of glucose; however, the accumulation of glycogen‐like glucose polymers can lead to degeneration and cellular death. Previously, we reported that the accumulation of glycogen in testis of transgenic animals overexpressing a constitutively active form of glycogen synthase enhances the apoptosis of pre‐meiotic male germ cells and a complete disorganization of the seminiferous tubules. Here we sought to further identify the effects of glycogen storage in cells from the seminiferous tubules and the mechanism behind the pro‐apoptotic activity induced by its accumulation. Using an in vitro culture of Sertoli cells (line 42GPA9) and spermatocyte‐like cells (line GC‐1) expressing a superactive form of glycogen synthase or the Protein Targeting to Glycogen (PTG), we found that glycogen synthesized in both cell lines is poorly branched. In addition, the immunodetection of key molecules of apoptotic events suggests that cellular death induced by polyglucosan molecules affects GC‐1 cells, but not 42GPA9 cells by mitochondrial impairment and activation of an intrinsic apoptotic pathway. Furthermore, we analyzed the effects of glycogen deposition during the establishment of an in vitro blood‐testis barrier. The results using a non‐permeable fluorescent molecule showed that, in conditions of over‐synthesis of glycogen, 42GPA9 cells do not lose their capacity to generate an impermeable barrier and the levels of connexin43, occludin, and ZO1 proteins were not affected. These results suggest that the accumulation of polyglucosan molecules has a selective effect—triggered by the intrinsic activation of the apoptotic pathway—in germ cells without directly affecting Sertoli cells. J. Cell. Physiol. 231: 2142–2152, 2016. © 2016 Wiley Periodicals, Inc.

  February 11, 2016   doi: 10.1002/jcp.25315   open full text
• Extracellular Matrix Stiffness Controls VEGF Signaling and Processing in Endothelial Cells.
  Kelsey D. Sack, Madelane Teran, Matthew A. Nugent.
  Journal of Cellular Physiology. February 08, 2016
  Vascular endothelial growth factor A (VEGF) drives endothelial cell maintenance and angiogenesis. Endothelial cell behavior is altered by the stiffness of the substrate the cells are attached to suggesting that VEGF activity might be influenced by the mechanical cellular environment. We hypothesized that extracellular matrix (ECM) stiffness modifies VEGF‐cell‐matrix tethering leading to altered VEGF processing and signaling. We analyzed VEGF binding, internalization, and signaling as a function of substrate stiffness in endothelial cells cultured on fibronectin (Fn) linked polyacrylamide gels. Cell produced extracellular matrices on the softest substrates were least capable of binding VEGF, but the cells exhibited enhanced VEGF internalization and signaling compared to cells on all other substrates. Inhibiting VEGF‐matrix binding with sucrose octasulfate decreased cell‐internalization of VEGF and, inversely, heparin pre‐treatment to enhance Fn‐matrix binding of VEGF increased cell‐internalization of VEGF regardless of matrix stiffness. β1 integrins, which connect cells to Fn, modulated VEGF uptake in a stiffness dependent fashion. Cells on hard surfaces showed decreased levels of activated β1 and inhibition of β1 integrin resulted in a greater proportional decrease in VEGF internalization than in cells on softer matrices. Extracellular matrix binding is necessary for VEGF internalization. Stiffness modifies the coordinated actions of VEGF‐matrix binding and β1 integrin binding/activation, which together are critical for VEGF internalization. This study provides insight into how the microenvironment may influence tissue regeneration and response to injury and disease. J. Cell. Physiol. 231: 2026–2039, 2016. © 2016 Wiley Periodicals, Inc.

  February 08, 2016   doi: 10.1002/jcp.25312   open full text
• The Outflow Pathway: A Tissue With Morphological and Functional Unity.
  Sergio Claudio Saccà, Stefano Gandolfi, Alessandro Bagnis, Gianluca Manni, Gianluca Damonte, Carlo Enrico Traverso, Alberto Izzotti.
  Journal of Cellular Physiology. February 08, 2016
  The trabecular meshwork (TM) plays an important role in high‐tension glaucomas. Indeed, the TM is a true organ, through which the aqueous humor flows from the anterior chamber to Schlemm's canal (SC). Until recently, the TM, which is constituted by endothelial‐like cells, was described as a kind of passive filter. In reality, it is much more. The cells delineating the structures of the collagen framework of the TM are endowed with a cytoskeleton, and are thus able to change their shape. These cells also have the ability to secrete the extracellular matrix, which expresses proteins and cytokines, and are capable of phagocytosis and autophagy. The cytoskeleton is attached to the nuclear membrane and can, in millionths of a second, send signals to the nucleus in order to alter the expression of genes in an attempt to adapt to biomechanical insult. Oxidative stress, as happens in aging, has a deleterious effect on the TM, leading eventually to cell decay, tissue malfunction, subclinical inflammation, changes in the extracellular matrix and cytoskeleton, altered motility, reduced outflow facility, and (ultimately) increased IOP. TM failure is the most relevant factor in the cascade of events triggering apoptosis in the inner retinal layers, including ganglion cells. J. Cell. Physiol. 231: 1876–1893, 2016. © 2016 Wiley Periodicals, Inc.

  February 08, 2016   doi: 10.1002/jcp.25305   open full text
• Expression of Ribosomal RNA and Protein Genes in Human Embryonic Stem Cells Is Associated With the Activating H3K4me3 Histone Mark.
  Sayyed K. Zaidi, Joseph R. Boyd, Rodrigo A. Grandy, Ricardo Medina, Jane B. Lian, Gary S. Stein, Janet L. Stein.
  Journal of Cellular Physiology. February 05, 2016
  Embryonic stem cells (ESCs) exhibit unrestricted and indefinite, but stringently controlled, proliferation, and can differentiate into any lineage in the body. In the current study, we test the hypothesis that expression of ribosomal RNA (rRNA) and ribosomal protein genes (RPGs) contribute to the ability of hESCs to proliferate indefinitely. Consistent with the accelerated growth rate of hESCs, we find that hESC lines H1 and H9 both exhibit significantly higher levels of rRNA when compared to a panel of normal and cancer human cell lines. Although many RPGs are expressed at levels that comparable to other human cell lines, a few RPGs also exhibit higher expression levels. In situ nuclear run‐on assays reveal that both nucleoli in hESCs actively transcribe nascent rRNA. Employing genome‐wide chromatin immunoprecipitation‐deep sequencing and bioinformatics approaches, we discovered that, RPGs are dominantly marked by the activating H3K4me3 histone mark in the G1, M, and G2 phases of the cell cycle. Interestingly, the rDNA repeats are marked by the activating H3K4me3 only in the M phase, and repressive H3K27me3 histone mark in all three cell cycle phases. Bioinformatics analyses also reveal that Myc, a known regulator of cell growth and proliferation, occupies both the rRNA genes and RPGs. Functionally, down‐regulation of Myc expression by siRNA results in a concomitant decrease in rRNA levels. Together, our results show that expression of rRNA, which is regulated by the Myc pluripotency transcription factor, and of RPGs in hESCs is associated with the activating H3K4me3 modification. J. Cell. Physiol. 231: 2007–2013, 2016. © 2016 Wiley Periodicals, Inc.

  February 05, 2016   doi: 10.1002/jcp.25309   open full text
• MicroRNA Regulation of Acute Lung Injury and Acute Respiratory Distress Syndrome.
  Subbiah Rajasekaran, Dhamotharan Pattarayan, P. Rajaguru, P. S. Sudhakar Gandhi, Rajesh K. Thimmulappa.
  Journal of Cellular Physiology. February 04, 2016
  The acute respiratory distress syndrome (ARDS), a severe form of acute lung injury (ALI), is a very common condition associated with critically ill patients, which causes substantial morbidity and mortality worldwide. Despite decades of research, effective therapeutic strategies for clinical ALI/ARDS are not available. In recent years, microRNAs (miRNAs), small non‐coding molecules have emerged as a major area of biomedical research as they post‐transcriptionally regulate gene expression in diverse biological and pathological processes, including ALI/ARDS. In this context, this present review summarizes a large body of evidence implicating miRNAs and their target molecules in ALI/ARDS originating largely from studies using animal and cell culture model systems of ALI/ARDS. We have also focused on the involvement of miRNAs in macrophage polarization, which play a critical role in regulating the pathogenesis of ALI/ARDS. Finally, the possible future directions that might lead to novel therapeutic strategies for the treatment of ALI/ARDS are also reviewed. J. Cell. Physiol. 231: 2097–2106, 2016. © 2016 Wiley Periodicals, Inc.

  February 04, 2016   doi: 10.1002/jcp.25316   open full text
• Differential Neuronal Plasticity of Dental Pulp Stem Cells From Exfoliated Deciduous and Permanent Teeth Towards Dopaminergic Neurons.
  Debanjana Majumdar, Mohammad Kanafi, Ramesh Bhonde, Pawan Gupta, Indrani Datta.
  Journal of Cellular Physiology. February 04, 2016
  Based on early occurrence in chronological age, stem‐cells from human exfoliated deciduous teeth (SHED) has been reported to possess better differentiation‐potential toward certain cell‐lineage in comparison to stem‐cells from adult teeth (DPSCs). Whether this same property between them extends for the yield of functional central nervous system neurons is still not evaluated. Hence, we aim to assess the neuronal plasticity of SHED in comparison to DPSCs toward dopaminergic‐neurons and further, if the difference is reflected in a differential expression of sonic‐hedgehog (SHH)‐receptors and basal‐expressions of tyrosine‐hydroxylase [TH; through cAMP levels]. Human SHED and DPSCs were exposed to midbrain‐cues [SHH, fibroblast growth‐factor8, and basic fibroblast growth‐factor], and their molecular, immunophenotypical, and functional characterization was performed at different time‐points of induction. Though SHED and DPSCs spontaneously expressed early‐neuronal and neural‐crest marker in their naïve state, only SHED expressed a high basal‐expression of TH. The upregulation of dopaminergic transcription‐factors Nurr1, Engrailed1, and Pitx3 was more pronounced in DPSCs. The yield of TH‐expressing cells decreased from 49.8% to 32.16% in SHED while it increased from 8.09% to 77.47% in DPSCs. Dopamine release and intracellular‐Ca2+ influx upon stimulation (KCl and ATP) was higher in induced DPSCs. Significantly lower‐expression of SHH‐receptors was noted in naïve SHED than DPSCs, which may explain the differential neuronal plasticity. In addition, unlike DPSCs, SHED showed a down‐regulation of cyclic adenosine‐monophosphate (cAMP) upon exposure to SHH; possibly another contributor to the lesser differentiation‐potential. Our data clearly demonstrates for the first time that DPSCs possess superior neuronal plasticity toward dopaminergic‐neurons than SHED; influenced by higher SHH‐receptor and lower basal TH expression. J. Cell. Physiol. 231: 2048–2063, 2016. © 2016 Wiley Periodicals, Inc.

  February 04, 2016   doi: 10.1002/jcp.25314   open full text
• Conversion of Prostate Adenocarcinoma to Small Cell Carcinoma‐Like by Reprogramming.
  Gisely T. Borges, Eneida F. Vêncio, Sue‐Ing Quek, Adeline Chen, Diego M. Salvanha, Ricardo Z.N. Vêncio, Holly M. Nguyen, Robert L. Vessella, Christopher Cavanaugh, Carol B. Ware, Pamela Troisch, Alvin Y. Liu.
  Journal of Cellular Physiology. February 04, 2016
  The lineage relationship between prostate adenocarcinoma and small cell carcinoma was studied by using the LuCaP family of xenografts established from primary neoplasm to metastasis. Expression of four stem cell transcription factor (TF) genes, LIN28A, NANOG, POU5F1, SOX2, were analyzed in the LuCaP lines. These genes, when force expressed in differentiated cells, can reprogram the recipients into stem‐like induced pluripotent stem (iPS) cells. Most LuCaP lines expressed POU5F1, while LuCaP 145.1, representative of small cell carcinoma, expressed all four. Through transcriptome database query, many small cell carcinoma genes were also found in stem cells. To test the hypothesis that prostate cancer progression from “differentiated” adenocarcinoma to “undifferentiated” small cell carcinoma could involve re‐expression of stem cell genes, the four TF genes were transduced via lentiviral vectors into five adenocarcinoma LuCaP lines—70CR, 73CR, 86.2, 92, 105CR—as done in iPS cell reprogramming. The resultant cells from these five transductions displayed a morphology of small size and dark appearing unlike the parentals. Transcriptome analysis of LuCaP 70CR* (“*” to denote transfected progeny) revealed a unique gene expression close to that of LuCaP 145.1. In a prostate principal components analysis space based on cell‐type transcriptomes, the different LuCaP transcriptome datapoints were aligned to suggest a possible ordered sequence of expression changes from the differentiated luminal‐like adenocarcinoma cell types to the less differentiated, more stem‐like small cell carcinoma types, and LuCaP 70CR*. Prostate cancer progression can thus be molecularly characterized by loss of differentiation with re‐expression of stem cell genes. J. Cell. Physiol. 231: 2040–2047, 2016. © 2016 Wiley Periodicals, Inc. Stem cell genes are re‐expressed in prostate cancer progression. Differentiated prostate adenocarcinoma could be converted to undifferentiated small cell carcinoma‐like by reprogramming.

  February 04, 2016   doi: 10.1002/jcp.25313   open full text
• Eriodictyol Inhibits RANKL‐Induced Osteoclast Formation and Function Via Inhibition of NFATc1 Activity.
  Fangming Song, Lin Zhou, Jinmin Zhao, Qian Liu, Mingli Yang, Renxiang Tan, Jun Xu, Ge Zhang, Julian M.W. Quinn, Jennifer Tickner, Yuanjiao Huang, Jiake Xu.
  Journal of Cellular Physiology. February 02, 2016
  Receptor activator of nuclear factor kappa‐B ligand (RANKL) induces differentiation and function of osteoclasts through triggering multiple signaling cascades, including NF‐κB, MAPK, and Ca2+‐dependent signals, which induce and activate critical transcription factor NFATc1. Targeting these signaling cascades may serve as an effective therapy against osteoclast‐related diseases. Here, by screening a panel of natural plant extracts with known anti‐inflammatory, anti‐tumor, or anti‐oxidant properties for possible anti‐osteoclastogenic activities we identified Eriodictyol. This flavanone potently suppressed RANKL‐induced osteoclastogenesis and bone resorption in a dose‐dependent manner without detectable cytotoxicity, suppressing RANKL‐induced NF‐κB, MAPK, and Ca2+ signaling pathways. Eriodictyol also strongly inhibited RANKL‐induction of c‐Fos levels (a critical component of AP‐1 transcription factor required by osteoclasts) and subsequent activation of NFATc1, concomitant with reduced expression of osteoclast specific genes including cathepsin K (Ctsk), V‐ATPase‐d2 subunit, and tartrate resistant acid phosphatase (TRAcP/Acp5). Taken together, these data provide evidence that Eriodictyol could be useful for the prevention and treatment of osteolytic disorders associated with abnormally increased osteoclast formation and function. J. Cell. Physiol. 231: 1983–1993, 2016. © 2016 Wiley Periodicals, Inc.

  February 02, 2016   doi: 10.1002/jcp.25304   open full text
• Prolonged Survival of Transplanted Osteoblastic Cells Does Not Directly Accelerate the Healing of Calvarial Bone Defects.
  Megumi Kitami, Masaru Kaku, Juan Marcelo Rosales Rocabado, Takako Ida, Nami Akiba, Katsumi Uoshima.
  Journal of Cellular Physiology. February 02, 2016
  Considering the increased interest in cell‐based bone regeneration, it is necessary to reveal the fate of transplanted cells and their substantive roles in bone regeneration. The aim of this study was to analyze the fate of transplanted cells and the effect of osteogenic cell transplantation on calvarial bone defect healing. An anti‐apoptotic protein, heat shock protein (HSP) 27, was overexpressed in osteoblasts. Then, the treated osteoblasts were transplanted to calvarial bone defect and their fate was analyzed to evaluate the significance of transplanted cell survival. Transient overexpression of Hsp27 rescued MC3T3‐E1 osteoblastic cells from H2O2‐induced apoptosis without affecting osteoblastic differentiation in culture. Transplantation of Hsp27‐overexpressing cells, encapsulated in collagen gel, showed higher proliferative activity, and fewer apoptotic cells in comparison with control cells. After 4‐week of transplantation, both control cell‐ and Hsp27 overexpressed cell‐transplanted groups showed significantly higher new bone formation in comparison with cell‐free gel‐transplantation group. Interestingly, the prolonged survival of transplanted osteoblastic cells by Hsp27 did not provide additional effect on bone healing. The transplanted cells in collagen gel survived for up to 4‐week but did not differentiate into bone‐forming osteoblasts. In conclusion, cell‐containing collagen gel accelerated calvarial bone defect healing in comparison with cell‐free collagen gel. However, prolonged survival of transplanted cells by Hsp27 overexpression did not provide additional effect. These results strongly indicate that cell transplantation‐based bone regeneration cannot be explained only by the increment of osteogenic cells. Further studies are needed to elucidate the practical roles of transplanted cells that will potentiate successful bone regeneration. J. Cell. Physiol. 231: 1974–1982, 2016. © 2016 Wiley Periodicals, Inc.

  February 02, 2016   doi: 10.1002/jcp.25302   open full text
• Improved Mitochondrial and Methylglyoxal‐Related Metabolisms Support Hyperproliferation Induced by 50 Hz Magnetic Field in Neuroblastoma Cells.
  Stefano Falone, Silvano Santini, Silvia di Loreto, Valeria Cordone, Marta Grannonico, Patrizia Cesare, Marisa Cacchio, Fernanda Amicarelli.
  Journal of Cellular Physiology. January 28, 2016
  Extremely low frequency magnetic fields (ELF‐MF) are common environmental agents that are suspected to promote later stages of tumorigenesis, especially in brain‐derived malignancies. Even though ELF magnetic fields have been previously linked to increased proliferation in neuroblastoma cells, no previous work has studied whether ELF‐MF exposure may change key biomolecular features, such as anti‐glycative defence and energy re‐programming, both of which are currently considered as crucial factors involved in the phenotype and progression of many malignancies. Our study investigated whether the hyperproliferation that is induced in SH‐SY5Y human neuroblastoma cells by a 50 Hz, 1 mT ELF magnetic field is supported by an improved defense towards methylglyoxal (MG), which is an endogenous cancer‐static and glycating α‐oxoaldehyde, and by rewiring of energy metabolism. Our findings show that not only the ELF magnetic field interfered with the biology of neuron‐derived malignant cells, by de‐differentiating further the cellular phenotype and by increasing the proliferative activity, but also triggered cytoprotective mechanisms through the enhancement of the defense against MG, along with a more efficient management of metabolic energy, presumably to support the rapid cell outgrowth. Intriguingly, we also revealed that the MF‐induced bioeffects took place after an initial imbalance of the cellular homeostasis, which most likely created a transient unstable milieu. The biochemical pathways and molecular targets revealed in this research could be exploited for future approaches aimed at limiting or suppressing the deleterious effects of ELF magnetic fields. J. Cell. Physiol. 231: 2014–2025, 2016. © 2016 Wiley Periodicals, Inc.

  January 28, 2016   doi: 10.1002/jcp.25310   open full text
• Signaling and Gene Regulatory Networks Governing Definitive Endoderm Derivation From Pluripotent Stem Cells.
  Abdulshakour Mohammadnia, Moein Yaqubi, Farzaneh Pourasgari, Eric Neely, Hossein Fallahi, Mohammad Massumi.
  Journal of Cellular Physiology. January 28, 2016
  The generation of definitive endoderm (DE) from pluripotent stem cells (PSCs) is a fundamental stage in the formation of highly organized visceral organs, such as the liver and pancreas. Currently, there is a need for a comprehensive study that illustrates the involvement of different signaling pathways and their interactions in the derivation of DE cells from PSCs. This study aimed to identify signaling pathways that have the greatest influence on DE formation using analyses of transcriptional profiles, protein–protein interactions, protein–DNA interactions, and protein localization data. Using this approach, signaling networks involved in DE formation were constructed using systems biology and data mining tools, and the validity of the predicted networks was confirmed experimentally by measuring the mRNA levels of hub genes in several PSCs‐derived DE cell lines. Based on our analyses, seven signaling pathways, including the BMP, ERK1‐ERK2, FGF, TGF‐beta, MAPK, Wnt, and PIP signaling pathways and their interactions, were found to play a role in the derivation of DE cells from PSCs. Lastly, the core gene regulatory network governing this differentiation process was constructed. The results of this study could improve our understanding surrounding the efficient generation of DE cells for the regeneration of visceral organs. J. Cell. Physiol. 231: 1994–2006, 2016. © 2016 Wiley Periodicals, Inc.

  January 28, 2016   doi: 10.1002/jcp.25308   open full text
• Effects of a Sublethal and Transient Stress of the Endoplasmic Reticulum on the Mitochondrial Population.
  Kayleen Vannuvel, Martine Van Steenbrugge, Catherine Demazy, Noëlle Ninane, Antoine Fattaccioli, Maude Fransolet, Patricia Renard, Martine Raes, Thierry Arnould.
  Journal of Cellular Physiology. January 15, 2016
  Endoplasmic reticulum (ER) and mitochondria are not discrete intracellular organelles but establish close physical and functional interactions involved in several biological processes including mitochondrial bioenergetics, calcium homeostasis, lipid synthesis, and the regulation of apoptotic cell death pathways. As many cell types might face a transient and sublethal ER stress during their lifetime, it is thus likely that the adaptive UPR response might affect the mitochondrial population. The aim of this work was to study the putative effects of a non‐lethal and transient endoplasmic reticulum stress on the mitochondrial population in HepG2 cells. The results show that thapsigargin and brefeldin A, used to induce a transient and sublethal ER stress, rapidly lead to the fragmentation of the mitochondrial network associated with a decrease in mitochondrial membrane potential, O2•− production and less efficient respiration. These changes in mitochondrial function are transient and preceded by the phosphorylation of JNK. Inhibition of JNK activation by SP600125 prevents the decrease in O2•− production and the mitochondrial network fragmentation observed in cells exposed to the ER stress but has no impact on the reduction of the mitochondrial membrane potential. In conclusion, our data show that a non‐lethal and transient ER stress triggers a rapid activation of JNK without inducing apoptosis, leading to the fragmentation of the mitochondrial network and a reduction of O2•− production. J. Cell. Physiol. 231: 1913–1931, 2016. © 2015 Wiley Periodicals, Inc.

  January 15, 2016   doi: 10.1002/jcp.25292   open full text
• Light/Dark Environmental Cycle Imposes a Daily Profile in the Expression of microRNAs in Rat CD133+ Cells.
  Marina Marçola, Camila M. Lopes‐Ramos, Eliana P. Pereira, Erika Cecon, Pedro A. Fernandes, Eduardo K. Tamura, Anamaria A. Camargo, Raphael B. Parmigiani, Regina P. Markus.
  Journal of Cellular Physiology. January 14, 2016
  The phenotype of primary cells in culture varies according to the donor environmental condition. We recently showed that the time of the day imposes a molecular program linked to the inflammatory response that is heritable in culture. Here we investigated whether microRNAs (miRNAs) would show differential expression according to the time when cells were obtained, namely daytime or nighttime. Cells obtained from explants of cremaster muscle and cultivated until confluence (∼20 days) presented high CD133 expression. Global miRNA expression analysis was performed through deep sequencing in order to compare both cultured cells. A total of 504 mature miRNAs were identified, with a specific miRNA signature being associated to the light versus dark phase of a circadian cycle. miR‐1249 and miR‐129‐2‐3p were highly expressed in daytime cells, while miR‐182, miR‐96‐5p, miR‐146a‐3p, miR‐146a‐5p, and miR‐223‐3p were highly expressed in nighttime cells. Nighttime cells are regulated for programs involved in cell processes and development, as well as in the inflammation, cell differentiation and maturation; while daytime cells express miRNAs that control stemness and cytoskeleton remodeling. In summary, the time of the day imposes a differential profile regarding to miRNA signature on CD133+ cells in culture. Understanding this daily profile in the phenotype of cultured cells is highly relevant for clinical outputs, including cellular therapy approaches. J. Cell. Physiol. 231: 1953–1963, 2016. © 2016 Wiley Periodicals, Inc.

  January 14, 2016   doi: 10.1002/jcp.25300   open full text
• Cell‐Autonomous Brown‐Like Adipogenesis of Preadipocytes From Retinoblastoma Haploinsufficient Mice.
  Petar D. Petrov, Andreu Palou, M. Luisa Bonet, Joan Ribot.
  Journal of Cellular Physiology. January 14, 2016
  Mechanisms behind the emergence of brown adipocyte‐like (brite or beige) adipocytes within white adipose tissue (WAT) are of interest. Retinoblastoma protein gene (Rb) haploinsufficiency associates in mice with improved metabolic regulation linked to a greater capacity for fatty acid oxidation and thermogenesis in WAT. We aimed to explain a feasible mechanism of WAT‐to‐BAT remodeling in this model. Differentiated primary adipocytes and Sca1‐positive preadipocytes derived from adipose depots of Rb+/− mice and wild‐type siblings were compared. Primary white Rb+/− adipocytes displayed under basal conditions increased glucose uptake and an enhanced expression of brown adipocyte‐related genes (Pparg, Ppargc1a, Ppargc1b, Prdm16, Cpt1b) but not of purported beige/brite transcriptional markers (Cd137, Tmem26, Tbx1, Slc27a1, Hoxc9, Shox2). Lack of induction of beige markers phenocopied results in WAT of adult Rb+/− mice. Flow cytometry analysis evidenced an increased number of preadipocytes in WAT depots of Rb+/− mice. Sca1+ preadipocytes from WAT of Rb+/− mice displayed increased gene expression of several transcription factors common to the brown and beige adipogenic programs (Prdm16, Pparg, Ppargc1a) and of receptors of bone morphogenetic proteins (BMPs); however, among the recently proposed beige markers, only Tbx1 was upregulated. Adult Rb+/− mice had increased circulating levels of BMP7. These results indicate that preadipose cells resident in WAT depots of Rb+/− mice retain an increased capacity for brown‐like adipogenesis that appears to be different from beige adipogenesis, and suggest that the contribution of these precursors to the Rb+/− adipose phenotype is driven, at least in part, by interaction with BMP7 pathways. J. Cell. Physiol. 231: 1941–1952, 2016. © 2016 Wiley Periodicals, Inc.

  January 14, 2016   doi: 10.1002/jcp.25299   open full text
• A NOXA/MCL‐1 Imbalance Underlies Chemoresistance of Malignant Rhabdoid Tumor Cells.
  Kazutaka Ouchi, Yasumichi Kuwahara, Tomoko Iehara, Mitsuru Miyachi, Yoshiki Katsumi, Kunihiko Tsuchiya, Eiichi Konishi, Akio Yanagisawa, Hajime Hosoi.
  Journal of Cellular Physiology. January 14, 2016
  Malignant rhabdoid tumor (MRT) is a rare aggressive pediatric cancer characterized by inactivation of SNF5, a core subunit of SWI/SNF complexes. Previously, we showed that SNF5 contributes to transcriptional activation of NOXA, a pro‐apoptotic protein that binds and inhibits the anti‐apoptotic protein MCL‐1. In this study, we found that NOXA expression was downregulated in MRT cell lines as well as in clinical MRT samples and that ectopically expressed NOXA bound MCL‐1 and increased the sensitivity of MRT cell lines to doxorubicin (DOX) by promoting apoptosis. Consistent with this finding, knockdown of MCL‐1 in MRT cell lines induced apoptosis and increased DOX sensitivity in MRT cells, and the MCL‐1 inhibitor TW‐37 synergized with DOX to induce MRT cell death. Our results suggest that modulation of the NOXA/MCL‐1 pathway may be a potential strategy for the treatment of patients with MRT. J. Cell. Physiol. 231: 1932–1940, 2016. © 2015 Wiley Periodicals, Inc. We show that NOXA expression is downregulated in malignant rhabdoid tumor (MRT) and that ectopic NOXA expression or knockdown of MCL‐1 induces apoptosis and increases DOX sensitivity in MRT cells. Moreover, the MCL‐1 inhibitor TW‐37 synergizes with DOX to induce MRT cell death. Our results suggest that modulation of the NOXA/MCL‐1 pathway may be a potential strategy for treatment of patients with MRT.

  January 14, 2016   doi: 10.1002/jcp.25293   open full text
• Vascular endothelial growth factor A regulates the secretion of different angiogenic factors in lung cancer cells.
  Daniela Frezzetti, Marianna Gallo, Cristin Roma, Amelia D'Alessio, Monica R. Maiello, Simona Bevilacqua, Nicola Normanno, Antonella De Luca.
  Journal of Cellular Physiology. November 06, 2015
  Vascular endothelial growth factor A (VEGFA) is one of the main mediators of angiogenesis in non‐small cell lung cancer (NSCLC). Recently, it has been described an autocrine feed‐forward loop in NSCLC cells in which tumor‐derived VEGFA promoted the secretion of VEGFA itself, amplifying the proangiogenic signal. In order to investigate the role of VEGFA in lung cancer progression, we assessed the effects of recombinant VEGFA on proliferation, migration and secretion of other angiogenic factors in A549, H1975 and HCC827 NSCLC cell lines. We found that VEGFA did not affect NSCLC cell proliferation and migration. On the other hand, we demonstrated that VEGFA not only produced a strong and persistent increase of VEGFA itself but also significantly induced the secretion of a variety of angiogenic factors, including follistatin (FST), hepatocyte growth factor (HGF), angiopoietin‐2 (ANGPT2), granulocyte‐colony stimulating factor (G‐CSF), interleukin (IL)‐8, leptin (LEP), platelet/endothelial cell adhesion molecule 1 (PECAM‐1) and platelet‐derived growth factor bb (PDGF‐BB). PI3K/AKT, RAS/ERK and STAT3 signalling pathways were found to mediate the effects of VEGFA in NSCLC cell lines. We also observed that VEGFA regulation mainly occurred at post‐transcriptional level and that NSCLC cells expressed different isoforms of VEGFA. Collectively, our data suggested that VEGFA contributes to lung cancer progression by inducing a network of angiogenic factors, which might offer potential for therapeutic intervention. This article is protected by copyright. All rights reserved

  November 06, 2015   doi: 10.1002/jcp.25243   open full text
• Complex‐I alteration and enhanced mitochondrial fusion are associated with prostate cancer progression.
  Julie V Philley, Anbarasu Kannan, Wenyi Qin, Edward R Sauter, Mitsuo Ikebe, Kate L Hertweck, Dean A Troyer, Oliver J Semmes, Santanu Dasgupta.
  Journal of Cellular Physiology. November 04, 2015
  Mitochondria (mt) encoded respiratory complex‐I (RCI) mutations and their pathogenicity remain largely unknown in prostate cancer (PCa). Little is known about the role of mtDNA loss on mt integrity in PCa. We determined mtDNA mutation in human and mice PCa and assessed the impact of mtDNA depletion on mt integrity. We also examined whether the circulating exosomes from PCa patients are transported to mt and carry mtDNA or mt proteins. We have employed next generation sequencing of the whole mt genome in human and Hi‐myc PCa. The impact of mtDNA depletion on mt integrity, presence of mtDNA and protein in sera exosomes was determined. A co‐culture of human PCa cells and the circulating exosomes followed by confocal imaging determined co‐localization of exosomes and mt. We observed frequent RCI mutations in human and Hi‐myc PCa which disrupted corresponding complex protein expression. Depletion of mtDNA in PCa cells influenced mt integrity, increased expression of MFN1, MFN2, PINK1 and decreased expression of MT‐TFA. Increased mt fusion and expression of PINK1 and DNM1L were also evident in the Hi‐myc tumors. RCI‐mtDNA, MFN2 and IMMT proteins were detected in the circulating exosomes of men with benign prostate hyperplasia (BPH) and progressive PCa. Circulating exosomes and mt co‐localized in PCa cells. Our study identified new pathogenic RCI mutations in PCa and defined the impact of mtDNA loss on mt integrity. Presence of mtDNA and mt proteins in the circulating exosomes implicated their usefulness for biomarker development. This article is protected by copyright. All rights reserved

  November 04, 2015   doi: 10.1002/jcp.25240   open full text
• Therapeutic Approaches in Mitochondrial Dysfunction, Proteolysis, and Structural Alterations of Diaphragm and Gastrocnemius in Rats With Chronic Heart Failure.
  Esther Barreiro, Ester Puig‐Vilanova, Judith Marin‐Corral, Alba Chacón‐Cabrera, Anna Salazar‐Degracia, Xavier Mateu, Luis Puente‐Maestu, Elena García‐Arumí, Antoni L. Andreu, Luis Molina.
  Journal of Cellular Physiology. November 04, 2015
  Patients with chronic heart failure (CHF) experience exercise intolerance, fatigue and muscle wasting, which negatively influence their survival. We hypothesized that treatment with either the antioxidant N‐acetyl cysteine (NAC) or the proteasome inhibitor bortezomib of rats with monocrotaline‐induced CHF may restore inspiratory and limb muscle mass, function, and structure through several molecular mechanisms involved in protein breakdown and metabolism in the diaphragm and gastrocnemius. In these muscles of CHF‐cachectic rats with and without treatment with NAC or bortezomib (n = 10/group) and non‐cachectic controls, proteolysis (tyrosine release, proteasome activities, ubiquitin‐proteasome markers), oxidative stress, inflammation, mitochondrial function, myosin, NF‐κB transcriptional activity, muscle structural abnormalities and fiber morphometry were analyzed together with muscle and cardiac functions. In diaphragm and gastrocnemius of CHF‐cachectic rats, tyrosine release, proteasome activity, protein ubiquitination, atrogin‐1, MURF‐1, NF‐κB activity, oxidative stress, inflammation and structural abnormalities were increased, while muscle and cardiac functions, myosin content, slow‐ and fast‐twitch fiber sizes, and mitochondrial activity were decreased. Concomitant treatment of CHF‐cachectic rats with NAC or bortezomib improved protein catabolism, oxidative stress, inflammation, muscle fiber sizes, function and damage, superoxide dismutase and myosin levels, mitochondrial function (complex I, gastrocnemius), cardiac function and decreased NF‐κB transcriptional activity in both muscles. Treatment of CHF‐cachectic animals with NAC or bortezomib attenuated the functional (heart, muscles), biological, and structural alterations in muscles. Nonetheless, future studies conducted in actual clinical settings are warranted in order to assess the potential beneficial effects and safety concerns of these pharmacological agents on muscle mass loss and wasting in CHF‐cachectic patients. This article is protected by copyright. All rights reserved

  November 04, 2015   doi: 10.1002/jcp.25241   open full text
• TNF‐α/TNFR2 Regulatory Axis Stimulates EphB2‐Mediated Neuroregeneration Via Activation of NF‐κB.
  Paul D. Pozniak, Armine Darbinyan, Kamel Khalili.
  Journal of Cellular Physiology. November 04, 2015
  HIV‐1 infected individuals are at high risk of developing HIV‐associated neurocognitive disorders (HAND) as HIV infection leads to neuronal injury and synaptic loss in the central nervous system (CNS). The neurotoxic effects of HIV‐1 are primarily a result of viral replication leading to the production of inflammatory chemokines and cytokines, including TNF‐α. Given an important role of TNF‐α in regulating synaptic plasticity, we investigated the effects of TNF‐α on the development of neuronal processes after mechanical injury, and we showed that TNF‐α treatment stimulates the regrowth of neuronal processes. To investigate transcriptional effects of TNF‐α on synaptic plasticity, we analyzed both human neurosphere and isolated neuronal cultures for the regulation of genes central to synaptic alterations during learning and memory. TNF‐α treatment upregulated Ephrin receptor B2 (EphB2), which is strongly involved in dendritic arborization and synaptic integrity. TNF‐α strongly activates the NF‐κB pathway, therefore, we propose that TNF‐α‐induced neurite regrowth occurs primarily through EphB2 signaling via stimulation of NF‐κB. EphB2 promoter activity increased with TNF‐α treatment and overexpression of NF‐κB. Direct binding of NF‐κB to the EphB2 promoter occurred in the ChIP assay, and site‐directed mutagenesis identified binding sites involved in TNF‐α‐induced EphB2 activation. TNF‐α induction of EphB2 was determined to occur specifically through TNF‐α receptor 2 (TNFR2) activation in human primary fetal neurons. Our observations provide a new avenue for the investigation on the impact of TNF‐α in the context of HIV‐1 neuronal cell damage as well as providing a potential therapeutic target in TNFR2 activation of EphB2. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  November 04, 2015   doi: 10.1002/jcp.25219   open full text
• Inhibition of polyamine uptake potentiates the anti‐proliferative effect of polyamine synthesis inhibition and preserves the contractile phenotype of vascular smooth muscle cells.
  Mario Grossi, Otto Phanstiel, Catarina Rippe, Karl Swärd, Azra Alajbegovic, Sebastian Albinsson, Amalia Forte, Lo Persson, Per Hellstrand, Bengt‐Olof Nilsson.
  Journal of Cellular Physiology. November 03, 2015
  Increased vascular smooth muscle cell (VSMC) proliferation is a factor in atherosclerosis and injury‐induced arterial (re)stenosis. Inhibition of polyamine synthesis by α‐difluoro‐methylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, attenuates VSMC proliferation with high sensitivity and specificity. However, cells can escape polyamine synthesis blockade by importing polyamines from the environment. To address this issue, polyamine transport inhibitors (PTIs) have been developed. We investigated the effects of the novel trimer44NMe (PTI‐1) alone and in combination with DFMO on VSMC polyamine uptake, proliferation and phenotype regulation. PTI‐1 efficiently inhibited polyamine uptake in primary mouse aortic and human coronary VSMCs in the absence as well as in the presence of DFMO. Interestingly, culture with DFMO for 2 days substantially (>95%) reduced putrescine (Put) and spermidine (Spd) contents without any effect on proliferation. Culture with PTI‐1 alone had no effect on either polyamine levels or proliferation rate, but the combination of both treatments reduced Put and Spd levels below the detection limit and inhibited proliferation. Treatment with DFMO for a longer time period (4 days) reduced Put and Spd below their detection limits and reduced proliferation, showing that only a small pool of polyamines is needed to sustain VSMC proliferation. Inhibited proliferation by polyamine depletion was associated with maintained expression of contractile smooth marker genes. In cultured intact mouse aorta, PTI‐1 potentiated the DFMO‐induced inhibition of cell proliferation. The combination of endogenous polyamine synthesis inhibition with uptake blockade is thus a viable approach for targeting unwanted vascular cell proliferation in vivo, including vascular restenosis. This article is protected by copyright. All rights reserved

  November 03, 2015   doi: 10.1002/jcp.25236   open full text
• Identification of a hematopoietic cell dedifferentiation‐inducing factor.
  Yunyuan Li, Hans Adomat, Emma Tomlinson Guns, Payman Hojabrpour, Vincent Duronio, Terry‐Ann Curran, Reza Baradar Jalili, William Jia, Zahid Delwar, Yun Zhang, Sanam Salimi Elizei, Aziz Ghahary.
  Journal of Cellular Physiology. November 03, 2015
  It has long been realized that hematopoietic cells may have the capacity to trans‐differentiate into non‐lymphohematopoietic cells under specific conditions. However, the mechanisms and the factors for hematopoietic cell trans‐differentiation remain unknown. In an in vitro culture system, we found that using a conditioned medium from proliferating fibroblasts can induce a subset of hematopoietic cells to become adherent fibroblast‐like cells (FLCs). FLCs are not fibroblasts nor other mesenchymal stromal cells, based on their expression of type‐1 collagen and other stromal cell marker genes. To identify the active factors in the conditioned medium, we cultured fibroblasts in a serum‐free medium and collected it for further purification. Using the fractions from filter devices of different molecular weight cut‐offs, and ammonium sulfate precipitation collected from the medium, we found the active fraction is a protein. We then purified this fraction by using fast protein liquid chromatography (FPLC) and identified it by mass spectrometer as macrophage colony‐stimulating factor (M‐CSF). The mechanisms of M‐CSF‐inducing trans‐differentiation of hematopoietic cells seem to involve a tyrosine kinase signalling pathway and its known receptor. The FLCs express a number of stem cell markers including SSEA‐1 and ‐3, OCT3/4, NANOG and SOX2. Spontaneous and induced differentiation experiments confirmed that FLCs can be further differentiated into cell types of three germ layers. These data indicate that hematopoietic cells can be induced by M‐CSF to dedifferentiate to multipotent stem cells. This study also provides a simple method to generate multipotent stem cells for clinical applications. This article is protected by copyright. All rights reserved

  November 03, 2015   doi: 10.1002/jcp.25239   open full text
• Interaction between Human Polyomavirus BK and Hypoxia inducible factor‐1 alpha.
  Lucia Signorini, Mattia Croci, Renzo Boldorini, Rafael Brandao Varella, Francesca Elia, Silvia Carluccio, Sonia Villani, Ramona Bella, Pasquale Ferrante, Serena Delbue.
  Journal of Cellular Physiology. November 03, 2015
  BK polyomavirus (BKV) has a worldwide seroprevalence of approximately 90%. After primary infection, BKV establishes a life‐long latency within the urogenital tract. The severe immunological impairment occurring in renal transplant recipients leads to BKV reactivation, which may result in Polyomavirus Associated Nephropathy (PVAN). While the transplanted kidney is transiently unperfused, Hypoxia Inducible Factors (HIFs) mediate the cellular response to hypoxia. The α‐subunit of HIF isoform 1 (HIF‐1α) may interact with several viruses, but until now, there has been no information regarding the interaction between BKV and HIF‐1α. The aim of this study is to investigate the possible interaction between HIF‐1α and BKV and its potential effect on the pathogenesis of PVAN. Screening of 17 kidney tissue samples revealed that HIF‐1α expression was 13.6‐fold higher in PVAN tissues compared to control tissues. A luminometric assay in co‐transfected African green monkey kidney cells (VERO) demonstrated BKV promoter activation ranging from 2‐ to 6‐fold (p < 0.05) when HIF‐1α was over‐expressed. A Chromatin ImmunoPrecipitation (ChIP) assay showed structural binding between the BKV promoter and HIF‐1α. The amount of BKV DNA increased by 3‐fold in VERO infected cells that were exposed to simulated hypoxia, compared to the cells not subjected to hypoxia. Both ex vivo and in vitro interactions between HIF‐1α and BKV were observed, suggesting that HIF‐1α, stabilized during transplantation, may be able to bind the BKV promoter and enhance BKV replication. Thus, hypoxia should be considered a risk factor for the development of PVAN in kidney transplant recipients. This article is protected by copyright. All rights reserved

  November 03, 2015   doi: 10.1002/jcp.25238   open full text
• Regulation and Functional Implications of Opioid Receptor Splicing in Opioid Pharmacology and HIV Pathogenesis.
  Patrick M. Regan, T. Dianne Langford, Kamel Khalili.
  Journal of Cellular Physiology. November 03, 2015
  Despite the identification and characterization of four opioid receptor subtypes and the genes from which they are encoded, pharmacological data does not conform to the predications of a four opioid receptor model. Instead, current studies of opioid pharmacology suggest the existence of additional receptor subtypes; however, no additional opioid receptor subtype has been identified to date. It is now understood that this discrepancy is due to the generation of multiple isoforms of opioid receptor subtypes. While several mechanisms are utilized to generate these isoforms, the primary mechanism involves alternative splicing of the pre‐mRNA transcript. Extensive alternative splicing patterns for opioid receptors have since been identified and discrepancies in opioid pharmacology are now partially attributed to variable expression of these isoforms. Recent studies have been successful in characterizing the localization of these isoforms as well as their specificity in ligand binding; however, the regulation of opioid receptor splicing specificity is poorly characterized. Furthermore, the functional significance of individual receptor isoforms and the extent to which opioid‐ and/or HIV‐mediated changes in the opioid receptor isoform profile contributes to altered opioid pharmacology or the well‐known physiological role of opioids in the exacerbation of HIV neurocognitive dysfunction is unknown. As such, the current review details constitutive splicing mechanisms as well as the specific architecture of opioid receptor genes, transcripts, and receptors in order to highlight the current understanding of opioid receptor isoforms, potential mechanisms of their regulation and signaling, and their functional significance in both opioid pharmacology and HIV‐associated neuropathology. This article is protected by copyright. All rights reserved

  November 03, 2015   doi: 10.1002/jcp.25237   open full text
• The Transcription Factor EB (TFEB) Regulates Osteoblast Differentiation through ATF4/CHOP‐Dependent Pathway.
  Erika Yoneshima, Kuniaki Okamoto, Eiko Sakai, Kazuhisa Nishishita, Noriaki Yoshida, Takayuki Tsukuba.
  Journal of Cellular Physiology. October 31, 2015
  Osteoblasts are bone‐forming cells that produce large amounts of collagen type I and various bone matrix proteins. Although osteoblast differentiation is highly regulated by various factors, it remains unknown whether lysosomes are directly involved in osteoblast differentiation. Here we demonstrate the transcription factor EB (TFEB), a master regulator of lysosomal biogenesis, modulates osteoblast differentiation. The expression levels of TFEB as well as those of endosomal/lysosomal proteins were up‐regulated during osteoblast differentiation using mouse osteoblastic MC3T3‐E1 cells. By gene knockdown (KD) experiments with small interfering RNA (siRNA), TFEB depletion caused markedly reduced osteoblast differentiation as compared with the control cells. Conversely, overexpression (OE) of TFEB resulted in strikingly enhanced osteoblastogenesis compared to the control cells. By analysis of down‐stream effector molecules, TFEB KD was found to cause marked up‐regulation of activating transcription factor4 (ATF4) and CCAAT/enhancer‐binding protein homologous protein (CHOP), both of which are essential factors for osteoblastogenesis. In contrast, TFEB OE promoted osteoblast differentiation through reduced expression of ATF4 and CHOP without differentiation agents. Given the importance of ATF4 and CHOP in osteoblastogenesis, it is clear that the TFEB‐regulated signaling pathway for osteoblast differentiation is involved in ATF4/CHOP‐dependent signaling pathway. This article is protected by copyright. All rights reserved

  October 31, 2015   doi: 10.1002/jcp.25235   open full text
• Ex Vivo Assay of Electrical Stimulation to Rat Sciatic Nerves: Cell Behaviors and Growth Factor Expression.
  Zhiyong Du, Olexandr Bondarenko, Dingkun Wang, Mahmoud Rouabhia, Ze Zhang.
  Journal of Cellular Physiology. October 30, 2015
  Neurite outgrowth and axon regeneration are known to benefit from electrical stimulation. However, how neuritis and their surroundings react to electrical field is difficult to replicate by monolayer cell culture. In this work freshly harvested rat sciatic nerves were cultured and exposed to two types of electrical field, after which time the nerve tissues were immunohistologically stained and the expression of neurotrophic factors and cytokines were evaluated. ELISA assay was used to confirm the production of specific proteins. All cell populations survived the 48h culture with little necrosis. Electrical stimulation was found to accelerate Wallerian degeneration and help Schwann cells to switch into migratory phenotype. Inductive electrical stimulation was shown to upregulate the secretion of multiple neurotrophic factors. Cellular distribution in nerve tissue was altered upon the application of an electrical field. This work thus presents an ex vivo model to study denervated axon in well controlled electrical field, bridging monolayer cell culture and animal experiment. It also demonstrated the critical role of electrical field distribution in regulating cellular activities. This article is protected by copyright. All rights reserved

  October 30, 2015   doi: 10.1002/jcp.25230   open full text
• Effects of Sleep Deprivation on Mice Bone Marrow and Spleen B Lymphopoiesis.
  Lisandro Lungato, Amanda Nogueira‐Pedro, Carolina Carvalho Dias, Edgar Julian Paredes‐Gamero, Sergio Tufik, Vânia D'Almeida.
  Journal of Cellular Physiology. October 30, 2015
  B lymphocytes are immune cells crucial for the maintenance and viability of the humoral response. Sleep is an essential event for the maintenance and integrity of all systems, including the immune system (IS). Thus, sleep deprivation (SD) causes problems in metabolism and homeostasis in many cell systems, including the IS. In this study, our goal was to determine changes in B lymphocytes from the bone marrow (BM) and spleen after SD. Three‐month‐old male Swiss mice were used. These mice were sleep deprived through the modified multiple platform method for different periods (24, 48, and 72 h), whereas another group was allowed to sleep for 24 h after 72 h of SD (rebound group) and a third group was allowed to sleep normally during the entire experiment. After this, the spleen and BM were collected, and cell analyses were performed. The numbers of B lymphocytes in the BM and spleen were reduced by SD. Additionally, reductions in the percentage of lymphocyte progenitors and their ability to form colonies were observed. Moreover, an increase in the death of B lymphocytes from the BM and spleen was associated with an increase in oxidative stress indicators, such as DCFH‐DA, CAT and mitochondrial SOD. Rebound was not able to reverse most of the alterations elicited by SD. The reduction in B lymphocytes and their progenitors by cell death, with a concomitant increase in oxidative stress, showed that SD promoted a failure in B lymphopoiesis. This article is protected by copyright. All rights reserved

  October 30, 2015   doi: 10.1002/jcp.25231   open full text
• cAMP and cGMP Play an Essential Role in Galvanotaxis of Cell Fragments.
  Kan Zhu, Yaohui Sun, Anh Miu, Michael Yen, Bowei Liu, Qunli Zeng, Alex Mogilner, Min Zhao.
  Journal of Cellular Physiology. October 30, 2015
  Cell fragments devoid of the nucleus and major organelles are found in physiology and pathology, for example platelets derived from megakaryocytes, and cell fragments from white blood cells and glioma cells. Platelets exhibit active chemotaxis. Fragments from white blood cells display chemotaxis, phagocytosis and bactericidal functions. Signaling mechanisms underlying migration of cell fragments are poorly understood. Here we used fish keratocyte fragments and demonstrated striking differences in signal transduction in migration of cell fragments and parental cells in a weak electric field. cAMP or cGMP agonists completely abolished directional migration of fragments, but had no effect on parental cells. The inhibition effects were prevented by pre‐incubating with cAMP and cGMP antagonists. Blocking cAMP and cGMP downstream signaling by inhibition of PKA and PKG also recovered fragment galvanotaxis. Both perturbations confirmed that the inhibitory effect was mediated by cAMP or cGMP signaling. Inhibition of cathode signaling with PI3K inhibitor LY294002 also prevented the effects of cAMP or cGMP agonists. Our results suggest that cAMP and cGMP are essential for galvanotaxis of cell fragments, in contrast to the signaling mechanisms in parental cells. This article is protected by copyright. All rights reserved

  October 30, 2015   doi: 10.1002/jcp.25229   open full text
• Mechanical Stimulation and IGF‐1 Enhance mRNA Translation Rate in Osteoblasts via Activation of the AKT‐mTOR Pathway.
  Astrid D. Bakker, Tom Gakes, Jolanda M.A. Hogervorst, Gerard M.J. de Wit, Jenneke Klein‐Nulend, Richard T. Jaspers.
  Journal of Cellular Physiology. October 27, 2015
  Insulin‐like growth factor‐1 (IGF‐1) is anabolic for muscle by enhancing the rate of mRNA translation via activation of AKT and subsequent activation of the mammalian target of rapamycin complex 1 (mTOR), thereby increasing cellular protein production. IGF‐1 is also anabolic for bone, but whether the mTOR pathway plays a role in the rate of bone matrix protein production by osteoblasts is unknown. We hypothesized that anabolic stimuli such as mechanical loading and IGF‐1 stimulate protein synthesis in osteoblasts via activation of the AKT‐mTOR pathway. MC3T3‐E1 osteoblasts were either or not subjected for 1 h to mechanical loading by pulsating fluid flow (PFF) or treated with or without human recombinant IGF‐1 (1‐100 ng/ml) for 0.5‐6 h, to determine phosphorylation of AKT and p70S6K (downstream of mTOR) by western blot. After 4 days of culture with or without the mTOR inhibitor rapamycin, total protein, DNA, and gene expression were quantified. IGF‐1 (100 ng/ml) reduced IGF‐1 gene expression, while PFF enhanced IGF‐1 expression. IGF‐1 did not affect collagen‐I gene expression. IGF‐1 dose‐dependently enhanced AKT and p70S6K phosphorylation at 2 and 6 h. PFF enhanced phosphorylation of AKT and p70S6K already within 1 h. Both IGF‐1 and PFF enhanced total protein per cell by ∼30%, but not in the presence of rapamycin. Our results show that IGF‐1 and PFF activate mTOR, thereby stimulating the rate of mRNA translation in osteoblasts. The known anabolic effect of mechanical loading and IGF‐1 on bone may thus be partly explained by mTOR‐mediated enhanced protein synthesis in osteoblasts. This article is protected by copyright. All rights reserved

  October 27, 2015   doi: 10.1002/jcp.25228   open full text
• Differential Regulation of Telomerase Reverse Transcriptase Promoter Activation and Protein Degradation by Histone Deacetylase Inhibition.
  Hua Qing, Jun Aono, Hannes M. Findeisen, Karrie L. Jones, Elizabeth B. Heywood, Dennis Bruemmer.
  Journal of Cellular Physiology. October 27, 2015
  Telomerase reverse transcriptase (TERT) maintains telomeres and is rate limiting for replicative life span. While most somatic tissues silence TERT transcription resulting in telomere shortening, cells derived from cancer or cardiovascular diseases express TERT and activate telomerase. In the present study, we demonstrate that histone deacetylase (HDAC) inhibition induces TERT transcription and promoter activation. At the protein level in contrast, HDAC inhibition decreases TERT protein abundance through enhanced degradation, which decreases telomerase activity and induces senescence. Finally, we demonstrate that HDAC inhibition decreases TERT expression during vascular remodeling in vivo. These data illustrate a differential regulation of TERT transcription and protein stability by HDAC inhibition and suggest that TERT may constitute an important target for the anti‐proliferative efficacy of HDAC inhibitors. This article is protected by copyright. All rights reserved

  October 27, 2015   doi: 10.1002/jcp.25226   open full text
• Effect of Docosahexaenoic Acid on Cell Cycle Pathways in Breast Cell Lines With Different Transformation Degree.
  Tania Rescigno, Anna Capasso, Mario Felice Tecce.
  Journal of Cellular Physiology. October 26, 2015
  n‐3 polyunsaturated fatty acids (PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), abundant in fish, have been shown to affect development and progression of some types of cancer, including breast cancer. The aim of our study was to further analyze and clarify the effects of these nutrients on the molecular mechanisms underlying breast cancer. Following treatments with DHA we examined cell viability, death, cell cycle, and some molecular effects in breast cell lines with different transformation, phenotypic, and biochemical characteristics (MCF‐10A, MCF‐7, SK‐BR‐3, ZR‐75‐1). These investigations showed that DHA is able to affect cell viability, proliferation, and cell cycle progression in a different way in each assayed breast cell line. The activation of ERK1/2 and STAT3 pathways and the expression and/or activation of molecules involved in cell cycle regulation such as p21Waf1/Cip1 and p53, are very differently regulated by DHA treatments in each cell model. DHA selectively: (i) arrests non tumoral MCF‐10A breast cells in G0/G1 cycle phase, activating p21Waf1/Cip1, and p53, (ii) induces to death highly transformed breast cells SK‐BR‐3, reducing ERK1/2 and STAT3 phosphorylation and (iii) only slightly affects each analyzed process in MCF‐7 breast cell line with transformation degree lower than SK‐BR‐3 cells. These findings suggest a more relevant inhibitory role of DHA within early development and late progression of breast cancer cell transformation and a variable effect in the other phases, depending on individual molecular properties and degree of malignancy of each clinical case. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  October 26, 2015   doi: 10.1002/jcp.25217   open full text
• Coordinated Dynamics of RNA Splicing Speckles in the Nucleus.
  Qiao Zhang, Krishna Kota Prasad, Samer G. Alam, Jeffrey A. Nickerson, Richard B. Dickinson, Tanmay P. Lele.
  Journal of Cellular Physiology. October 23, 2015
  Despite being densely packed with chromatin, nuclear bodies and a nucleoskeletal network, the nucleus is a remarkably dynamic organelle(Hübner et al., 2013). Chromatin loops form and relax, RNA transcripts and transcription factors move diffusively, and nuclear bodies move. We show here that RNA splicing speckled domains (splicing speckles) fluctuate in constrained nuclear volumes and remodel their shapes. Small speckles move in a directed way toward larger speckles with which they fuse. The movement is reduced upon decreasing cellular ATP levels, moderately reduced after inhibition of SWI/SNF chromatin remodeling and modestly increased upon inhibiting RNA polymerase II activity. To define the paths through which speckles can translocate in the nucleus, we generated a pressure gradient to create flows in the nucleus. In response to the pressure gradient, speckles moved in a coordinated way along curvilinear paths in the nucleus. Collectively, our results demonstrate a new type of ATP‐dependent motion in the nucleus. We present a model where recycling splicing factors return as part of small sub‐speckles from distal sites of RNA processing to larger splicing speckles a directed ATP‐driven mechanism through interchromatin spaces. This article is protected by copyright. All rights reserved

  October 23, 2015   doi: 10.1002/jcp.25224   open full text
• Common and Rare Variants Associated with Alzheimer's Disease.
  Hany Marei, Asmaa Althani, Mohamed El Zowalaty, Mohammad A. Albanna, Carlo Cenciarelli, Tengfei Wang, Thomas Caceci.
  Journal of Cellular Physiology. October 23, 2015
  Alzheimer's disease (AD) is one of the most devastating disorder. Despite the continuing increase of its incidence among aging population, no effective cure has been developed mainly due to difficulties in early diagnosis of the disease before damaging of the brain, and the failure to explore its complex underlying molecular mechanisms. Recent technological advances in genome‐wide association studies (GWAS) and high throughput next generation whole genome, and exome sequencing had deciphered many of AD‐related loci, and discovered single nucleotide polymorphisms (SNPs) that are associated with altered AD molecular pathways. Highlighting altered molecular pathways linked to AD pathogenesis is crucial to identify novel diagnostic and therapeutic AD targets. This article is protected by copyright. All rights reserved

  October 23, 2015   doi: 10.1002/jcp.25225   open full text
• Acetylcholine Attenuates Hypoxia/Reoxygenation Injury by Inducing Mitophagy Through PINK1/Parkin Signal Pathway in H9c2 Cells.
  Lei Sun, Mei Zhao, Yang Yang, Run‐Qing Xue, Xiao‐Jiang Yu, Jian‐Kang Liu, Wei‐Jin Zang.
  Journal of Cellular Physiology. October 23, 2015
  Acetylcholine (ACh) protected against cardiac injury via promoting autophagy and mitochondrial biogenesis, however, the involvement of mitophagy in ACh‐elicited cardioprotection remains unknown. In the present study, H9c2 cardiomyocytes were subjected to hypoxia/reoxygenation (H/R) and ACh treatment during reoxygenation. Mitophagy markers PTEN‐induced kinase 1 (PINK1) and Parkin translocation were examined using western blot and confocal fluorescence microscopy. Mitochondrial membrane potential and reactive oxygen species (ROS) were detected with fluorescence staining. We found that H/R‐treated cells exhibited reduced levels of PINK1 and Parkin in mitochondria, accompanied with decreased autophagy flux (reduced LC3‐II/LC3‐I and increased p62). Conversely, ACh increased PINK1 and Parkin translocation to mitochondria and enhanced autophagy proteins. Confocal imaging of Parkin and MitoTracker Green‐labeled mitochondria further confirmed ACh‐induced mitochondrial translocation of Parkin, which was reversed by M2 receptor antagonist methoctramine and M2 receptor siRNA, suggesting ACh could induce mitophagy by M2 receptor after H/R. Mitophagy inhibitor 3‐methaladenine abolished ACh‐induced mitoprotection, manifesting as aggravated mitochondrial morphology disruption, ATP and membrane potential depletion, increased ROS overproduction, and apoptosis. Furthermore, PINK1/Parkin siRNA attenuated the protective effects of ACh against ATP loss and oxidative stress due to mitochondrial‐dependent injury. Taken together, ACh promoted mitochondrial translocation of PINK1/Parkin to stimulate cytoprotective mitophagy via M2 receptor, which may provide beneficial targets in the preservation of cardiac homeostasis against H/R injury. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  October 23, 2015   doi: 10.1002/jcp.25215   open full text
• Mitochondrial Dysfunction and Ca2+ Overload Contributes to Hesperidin Induced Paraptosis in Hepatoblastoma Cells, HepG2.
  Silvia Yumnam, Gyeong Eun Hong, Suchismita Raha, Venu Venkatarame Gowda Saralamma, Ho Jeong Lee, Won‐Sup Lee, Eun‐Hee Kim, Gon Sup Kim.
  Journal of Cellular Physiology. October 22, 2015
  Paraptosis is a programmed cell death which is morphologically and biochemically different from apoptosis. In this study, we have investigated the role of Ca2+ in hesperidin‐induced paraptotic cell death in HepG2 cells. Increase in mitochondrial Ca2+ level was observed in hesperidin treated HepG2 cells but not in normal liver cancer cells. Inhibition of inositol‐1,4,5‐triphosphate receptor (IP3) and ryanodine receptor also block the mitochondrial Ca2+ accumulation suggesting that the release of Ca2+ from the endoplasmic reticulum (ER) may probably lead to the increase in mitochondrial Ca2+ level. Pretreatment with ruthenium red (RuRed), a Ca2+ uniporter inhibitor inhibited the hesperidin‐induced mitochondrial Ca2+ overload, swelling of mitochondria and cell death in HepG2 cells. It has also been demonstrated that mitochondrial Ca2+ influxes act upstream of ROS and mitochondrial superoxide production. The increased ROS production further leads to mitochondrial membrane loss in hesperidin treated HepG2 cells. Taken together our results show that IP3R and ryanodine receptor mediated release of Ca2+ from the ER and its subsequent influx through the uniporter into mitochondria contributes to hesperidin‐induced paraptosis in HepG2 cells. This article is protected by copyright. All rights reserved

  October 22, 2015   doi: 10.1002/jcp.25222   open full text
• Different Blood‐Borne Human Osteoclast Precursors Respond in Distinct Ways to IL‐17A.
  Sara Sprangers, Ton Schoenmaker, Yixuan Cao, Vincent Everts, Teun J. de Vries.
  Journal of Cellular Physiology. October 22, 2015
  Osteoclasts are bone‐degrading cells that are formed through fusion of their monocytic precursors. Three distinct subsets of monocytes have been identified in human peripheral blood: classical, intermediate and non‐classical monocytes. They are known to play different roles in physiology and pathology, but their capacity to differentiate into osteoclasts and whether inflammatory cytokines influence this differentiation is unknown. We hypothesized that classical, intermediate and non‐classical monocytes generate functionally different osteoclasts and that they respond in different ways to the inflammatory cytokine interleukin‐17A (IL‐17A). To investigate this, the different monocyte subsets were isolated from human peripheral blood and osteoclastogenesis was induced with the cytokines M‐CSF and RANKL, with or without IL‐17A. We found that all subsets are able to differentiate into osteoclasts in vitro, and that both osteoclastogenesis and subsequent bone resorption was distinctly affected by IL‐17A. Osteoclastogenesis and bone resorption by osteoclasts derived from classical monocytes remained unaffected by IL‐17A, while osteoclast formation from intermediate monocytes was inhibited by the cytokine. Surprisingly, bone resorption by osteoclasts derived from intermediate monocytes remained at similar levels as control cultures, indicating an increased bone resorbing activity by these osteoclasts. Limited numbers of osteoclasts were formed from non‐classical monocytes on bone and no bone resorption was detected, which suggest that these cells belong to a cell lineage different from the osteoclast. By providing more insight into osteoclast formation from human blood monocytes, this study contributes to the possible targeting of specific osteoclast precursors as a therapeutic approach for diseases associated with inflammatory bone loss. This article is protected by copyright. All rights reserved

  October 22, 2015   doi: 10.1002/jcp.25220   open full text
• Selective Life‐Long Skeletal Myofiber—Targeted VEGF Gene Ablation Impairs Exercise Capacity in Adult Mice.
  Kechun Tang, Yusu Gu, Nancy D. Dalton, Harrieth Wagner, Kirk L. Peterson, Peter D. Wagner, Ellen C. Breen.
  Journal of Cellular Physiology. October 22, 2015
  Exercise is dependent on adequate oxygen supply for mitochondrial respiration in both cardiac and locomotor muscle. To determine whether skeletal myofiber VEGF is critical for regulating exercise capacity, independent of VEGF function in the heart, ablation of the VEGF gene was targeted to skeletal myofibers (skmVEGF−/−) during embryogenesis (∼ E9.5), leaving intact VEGF expression by all other cells in muscle. In adult mice, VEGF levels were decreased in the soleus (by 65%), plantaris (94%), gastrocnemius (74%), EDL (99%) and diaphragm (64%) (P < 0.0001, each muscle). VEGF levels were unchanged in the heart. Treadmill speed (WT 86 ± 4 cm/sec, skmVEGF−/− 70 ± 5 cm/sec, P = 0.006) and endurance (WT 78 ± 24 min, skmVEGF−/− 18 ± 4 min, P = 0.0004) were severely limited in skmVEGF−/− mice in contrast to minor effect of conditional skmVEGF gene deletion in the adult. Body weight was also reduced (WT 22.8 ± 1.6 g, skmVEGF−/−, 21.1 ± 1.5, P = 0.02), but the muscle mass/body weight ratio was unchanged. The capillary/fiber ratio was lower in skmVEGF−/− plantaris (WT 1.51 ± 0.12, skmVEGF−/− 1.16 ± 0.20, P = 0.01), gastrocnemius (WT 1.61 ± 0.08, skmVEGF−/− 1.39 ± 0.08, P = 0.01), EDL (WT 1.36 ± 0.07, skmVEGF−/− 1.14 ± 0.13, P = 0.03) and diaphragm (WT 1.39 ±  0.18, skmVEGF−/− 0.79 ± 0.16, P = 0.0001) but, not in soleus. Cardiac function (heart rate, maximal pressure, maximal dP/dt, minimal dP/dt,) in response to dobutamine was not impaired in anesthetized skmVEGF−/− mice. Isolated soleus and EDL fatigue times were 16% and 20% (P < 0.02) longer, respectively, in skmVEGF−/− mice than the WT group. These data suggest that skeletal myofiber VEGF expressed during development is necessary to establish capillary networks that allow maximal exercise capacity. J. Cell. Physiol. 231: 505–511, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25097   open full text
• D‐Aspartate Induces Proliferative Pathways in Spermatogonial GC‐1 Cells.
  Alessandra Santillo, Sara Falvo, Paolo Chieffi, Maria Maddalena Di Fiore, Rosalba Senese, Gabriella Chieffi Baccari.
  Journal of Cellular Physiology. October 22, 2015
  D‐aspartate (D‐Asp) is an endogenous amino acid present in vertebrate tissues, with particularly high levels in the testis. In vivo studies indicate that D‐Asp indirectly stimulates spermatogenesis through the hypothalamic‐pituitary‐gonadal axis. Moreover, in vitro studies have demonstrated that D‐Asp up‐regulates testosterone production in Leydig cells by enhancing expression of the steroidogenic acute regulatory protein. In this study, a cell line derived from immortalized type‐B mouse spermatogonia retaining markers of mitotic germ cells (GC‐1) was employed to explore more direct involvement of D‐Asp in spermatogenesis. Activity and protein expression of markers of cell proliferation were determined at intervals during incubation in D‐Asp‐containing medium. D‐Asp induced phosphorylation of ERK and Akt proteins, stimulated expression of PCNA and Aurora B, and enhanced mRNA synthesis and protein expression of P450 aromatase and protein expression of Estrogen Receptor β (ERβ). These results are the first demonstration of a direct effect of D‐Asp on spermatogonial mitotic activity. Considering that spermatogonia express the NR1 subunit of the N‐Methyl‐D‐Aspartic Acid receptor (NMDAR), we suggest that their response to D‐Asp depends on NMDAR‐mediated activation of the ERK and Akt pathways and is further enhanced by activation of the P450 aromatase/ERβ pathway. J. Cell. Physiol. 231: 490–495, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25095   open full text
• Hydroxytyrosol Inhibits Cannabinoid CB1 Receptor Gene Expression in 3T3‐L1 Preadipocyte Cell Line.
  Valeria Tutino, Antonella Orlando, Francesco Russo, Maria Notarnicola.
  Journal of Cellular Physiology. October 22, 2015
  The 3T3‐L1 preadipocyte cell line is a well characterized cell model for studying the adipocyte status and the molecular mechanisms involved in differentiation of these cells. 3T3‐L1 preadipocytes have the ability to synthesize and degrade endocannabinoid anandamide (AEA) and their differentiation into adipocytes increases the expression of cannabinoid (CB1) and PPAR‐γ receptors. Clinically, the blocking stimulation of the endocannabinoid pathway has been one of the first approaches proposed to counteract the obesity and obesity‐associated diseases (such as diabetes, metabolic syndrome and cancer). In this connection, here we studied in cultured 3T3‐L1 pre‐adipocytes the effects of n‐3‐PUFA, α‐Linolenic acid (OM‐3), n‐6‐PUFA, Linoleic acid (OM‐6), and hydroxytyrosol (HT) on the expression of CB1 receptor gene and the adipogenesis‐related genes PPAR‐γ, Fatty Acid Synthase (FAS) and Lipoprotein Lipase (LPL). HT was able to inhibit 3T3‐L1 cell differentiation by down‐regulating cell proliferation and CB1 receptor gene expression. HT exhibited anti‐adipogenic effects, whereas OM‐3 and OM‐6 exerted an inhibitory action on cell proliferation associated with an induction of the preadipocytes differentiation and CB1 receptor gene expression. Moreover, the expression of FAS and LPL genes resulted increased after treatment with both HT and OM‐3 and OM‐6. The present study points out that the intake of molecules such as HT, contained in extra virgin olive oil, may be considered also in view of antiobesity and antineoplastic properties by acting directly on the adipose tissue and modulating CB1 receptor gene transcription. J. Cell. Physiol. 231: 483–489, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25094   open full text
• Nitric Oxide Up‐Regulates RUNX2 in LNCaP Prostate Tumours: Implications for Tumour Growth In Vitro and In Vivo.
  Heather Nesbitt, Gillian Browne, Katie M. O'Donovan, Niall M. Byrne, Jenny Worthington, Stephanie R. McKeown, Declan J. McKenna.
  Journal of Cellular Physiology. October 22, 2015
  Aberrant expression of the transcription factor RUNX2 in prostate cancer has a number of important consequences including increased resistance to apoptosis, invasion and metastasis to bone. We previously demonstrated that hypoxia up‐regulated RUNX2 in tumour cells, which in turn up‐regulated the anti‐apoptotic factor Bcl‐2. Here, we investigate the impact of nitric oxide (NO) on RUNX2 and Bcl‐2 expression in prostate cancer and further, how RUNX2 over‐expression can impact tumour growth, angiogenesis and oxygenation in vivo. The effect of NO levels on RUNX2 and thus Bcl‐2 expression was examined in prostate cancer cells in vitro using methods including gene and protein expression analyses, nitrite quantitation, protein‐DNA interaction assays (ChIP) and viability assays (XTT). The effect of RUNX2 over‐expression on tumour physiology (growth, oxygenation and angiogenesis) was also assessed in vivo using LNCaP xenografts. A low (but not high) concentration of NO (10 μM) induced expression of RUNX2 and Bcl‐2, conferring resistance to docetaxel. These effects were induced via the ERK and PI3K pathways and were dependent on intact AP‐1 binding sites in the RUNX2 promoter. RUNX2 over‐expression in LNCaP tumours in vivo decreased the time to tumour presentation and increased tumour growth. Moreover, these tumours exhibited improved tumour angiogenesis and oxygenation. Low levels of NO increase expression of RUNX2 and Bcl‐2 in LNCaP prostate tumour cells, and in vivo up‐regulation of RUNX2 created tumours with a more malignant phenotype. Collectively, our data reveals the importance of NO‐regulation of key factors in prostate cancer disease progression. J. Cell. Physiol. 231: 473–482, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25093   open full text
• Intravenous Immunoglobulin (IVIG) Attenuates TNF‐Induced Pathologic Bone Resorption and Suppresses Osteoclastogenesis by Inducing A20 Expression.
  Min Joon Lee, Elisha Lim, Se‐Hwan Mun, Seyeon Bae, Koichi Murata, Lionel B. Ivashkiv, Kyung‐Hyun Park‐Min.
  Journal of Cellular Physiology. October 22, 2015
  Investigations on the therapeutic effects of intravenous immunoglobulin (IVIG) have focused on the suppression of autoantibody and immune complex‐mediated inflammatory pathogenesis. Inflammatory diseases such as rheumatoid arthritis are often accompanied by excessive bone erosion but the effect of IVIG on osteoclasts, bone‐resorbing cells, has not been studied. Here, we investigate whether IVIG directly regulates osteoclast differentiation and has therapeutic potential for suppressing osteoclast‐mediated pathologic bone resorption. IVIG or cross‐linking of Fcγ receptors with plate‐bound IgG suppressed receptor activator of nuclear factor‐κ B ligand (RANKL)‐induced osteoclastogenesis and expression of osteoclast‐related genes such as integrin β3 and cathepsin K in a dose‐dependent manner. Mechanistically, IVIG or plate‐bound IgG suppressed osteoclastogenesis by downregulating RANKL‐induced expression of NFATC1, the master regulator of osteoclastogenesis. IVIG suppressed NFATC1 expression by attenuating RANKL‐induced NF‐κB signaling, explained in part by induction of the inflammatory signaling inhibitor A20. IVIG administration attenuated in vivo osteoclastogenesis and suppressed bone resorption in the tumor necrosis factor (TNF)‐induced calvarial osteolysis model. Our findings show that, in addition to suppressing inflammation, IVIG directly inhibits osteoclastogenesis through a mechanism involving suppression of RANK signaling. Direct suppression of osteoclast differentiation may provide beneficial effects on preserving bone mass when IVIG is used to treat rheumatic disorders. J. Cell. Physiol. 231: 449–458, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25091   open full text
• Distinct Roles for Intestinal Epithelial Cell‐Specific Hdac1 and Hdac2 in the Regulation of Murine Intestinal Homeostasis.
  Alexis Gonneaud, Naomie Turgeon, François Boudreau, Nathalie Perreault, Nathalie Rivard, Claude Asselin.
  Journal of Cellular Physiology. October 22, 2015
  The intestinal epithelium responds to and transmits signals from the microbiota and the mucosal immune system to insure intestinal homeostasis. These interactions are in part conveyed by epigenetic modifications, which respond to environmental changes. Protein acetylation is an epigenetic signal regulated by histone deacetylases, including Hdac1 and Hdac2. We have previously shown that villin‐Cre‐inducible intestinal epithelial cell (IEC)‐specific Hdac1 and Hdac2 deletions disturb intestinal homeostasis. To determine the role of Hdac1 and Hdac2 in the regulation of IEC function and the establishment of the dual knockout phenotype, we have generated villin‐Cre murine models expressing one Hdac1 allele without Hdac2, or one Hdac2 allele without Hdac1. We have also investigated the effect of short‐term deletion of both genes in naphtoflavone‐inducible Ah‐Cre and tamoxifen‐inducible villin‐CreER mice. Mice with one Hdac1 allele displayed normal tissue architecture, but increased sensitivity to DSS‐induced colitis. In contrast, mice with one Hdac2 allele displayed intestinal architecture defects, increased proliferation, decreased goblet cell numbers as opposed to Paneth cells, increased immune cell infiltration associated with fibrosis, and increased sensitivity to DSS‐induced colitis. In comparison to dual knockout mice, intermediary activation of Notch, mTOR, and Stat3 signaling pathways was observed. While villin‐CreER Hdac1 and Hdac2 deletions led to an impaired epithelium and differentiation defects, Ah‐Cre‐mediated deletion resulted in blunted proliferation associated with the induction of a DNA damage response. Our results suggest that IEC determination and intestinal homeostasis are highly dependent on Hdac1 and Hdac2 activity levels, and that changes in the IEC acetylome may alter the mucosal environment. J. Cell. Physiol. 231: 436–448, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25090   open full text
• S‐Adenosylmethionine Affects ERK1/2 and Stat3 Pathways and Induces Apotosis in Osteosarcoma Cells.
  Concetta Paola Ilisso, Luigi Sapio, Donatella Delle Cave, Michela Illiano, Annamaria Spina, Giovanna Cacciapuoti, Silvio Naviglio, Marina Porcelli.
  Journal of Cellular Physiology. October 22, 2015
  Osteosarcoma is a very aggressive bone tumor. Its clinical outcome remains discouraging despite intensive surgery, radiotherapy, and chemotherapy. Thus, novel therapeutic approaches are demanded. S‐Adenosylmethionine (AdoMet) is a naturally occurring molecule that is synthesized in our body by methionine adenosyltransferase isoenzymes and is also available as a nutritional supplement. AdoMet is the principal methyl donor in numerous methylation reactions and is involved in many biological functions. Interestingly, AdoMet has been shown to exert antiproliferative action in various cancer cells. However, the underlying molecular mechanisms are just starting to be studied. Here, we investigated the effects of AdoMet on the proliferation of osteosarcoma U2OS cells and the underlying mechanisms. We carried out direct cell number counting, MTT and flow cytometry‐based assays, and immunoblotting experiments in response to AdoMet treatment. We found that AdoMet strongly inhibits proliferation of U2OS cells by slowing‐down cell cycle progression and by inducing apoptosis. We also report that AdoMet consistently causes an increase of p53 and p21 cell‐cycle inhibitor, a decrease of cyclin A and cyclin E protein levels, and a marked increase of pro‐apoptotic Bax/Bcl‐2 ratio, with caspase‐3 activation and PARP cleavage. Moreover, the AdoMet‐induced antiproliferative effects were dynamically accompanied by profound changes in ERK1/2 and STAT3 protein and phosphorylation levels. Altogether, our data enforce the evidence of AdoMet acting as a biomolecule with antiproliferative action in osteosarcoma cells, capable of down‐regulating ERK1/2 and STAT3 pathways leading to cell cycle inhibition and apoptosis, and provide a rationale for the possible use of AdoMet in osteosarcoma therapy. J. Cell. Physiol. 231: 428–435, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25089   open full text
• Grp78 Is Critical for Amelogenin‐Induced Cell Migration in a Multipotent Clonal Human Periodontal Ligament Cell Line.
  Kyosuke Toyoda, Takao Fukuda, Terukazu Sanui, Urara Tanaka, Kensuke Yamamichi, Ryo Atomura, Hidefumi Maeda, Atsushi Tomokiyo, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura.
  Journal of Cellular Physiology. October 22, 2015
  Periodontal ligament stem cells (PDLSCs) are known to play a pivotal role in regenerating the periodontium. Amelogenin, which belongs to a family of extracellular matrix (ECM) proteins, is a potential bioactive molecule for periodontal regenerative therapy. However, its downstream target molecules and/or signaling patterns are still unknown. Our recent proteomic study identified glucose‐regulated protein 78 (Grp78) as a new amelogenin‐binding protein. In this study, we demonstrate, for the first time, the cellular responses induced by the biological interaction between amelogenin and Grp78 in the human undifferentiated PDL cell line 1‐17, which possesses the most typical characteristics of PDLSCs. Confocal co‐localization experiments revealed the internalization of recombinant amelogenin (rM180) via binding to cell surface Grp78, and the endocytosis was inhibited by the silencing of Grp78 in 1‐17 cells. Microarray analysis indicated that rM180 and Grp78 regulate the expression profiles of cell migration‐associated genes in 1‐17 cells. Moreover, Grp78 overexpression enhanced rM180‐induced cell migration and adhesion without affecting cell proliferation, while silencing of Grp78 diminished these activities. Finally, binding of rM180 to Grp78 promoted the formation of lamellipodia, and the simultaneous activation of Rac1 was also demonstrated by NSC23766, a widely accepted Rac1 inhibitor. These results suggest that Grp78 is essential for enhancing amelogenin‐induced migration in 1‐17 cells. The biological interaction of amelogenin with Grp78 offers significant therapeutic potential for understanding the biological components and specific functions involved in the signal transduction of amelogenin‐induced periodontal tissue regeneration. J. Cell. Physiol. 231: 414–427, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25087   open full text
• TRPV3 Channel Negatively Regulates Cell Cycle Progression and Safeguards the Pluripotency of Embryonic Stem Cells.
  Iek Chi Lo, Hing Chung Chan, Zenghua Qi, Kwun Lam Ng, Chun So, Suk Ying Tsang.
  Journal of Cellular Physiology. October 22, 2015
  Embryonic stem cells (ESCs) have tremendous potential for research and future therapeutic purposes. However, the calcium handling mechanism in ESCs is not fully elucidated. Aims of this study are (1) to investigate if transient receptor potential vanilloid‐3 (TRPV3) channels are present in mouse ESCs (mESCs) and their subcellular localization; (2) to investigate the role of TRPV3 in maintaining the characteristics of mESCs. Western blot and immunocytochemistry showed that TRPV3 was present at the endoplasmic reticulum (ER) of mESCs. Calcium imaging showed that, in the absence of extracellular calcium, TRPV3 activators camphor and 6‐tert‐butyl‐m‐cresol increased the cytosolic calcium. However, depleting the ER store in advance of activator addition abolished the calcium increase, suggesting that TRPV3 released calcium from the ER. To dissect the functional role of TRPV3, TRPV3 was activated and mESC proliferation was measured by trypan blue exclusion and MTT assays. The results showed that TRPV3 activation led to a decrease in mESC proliferation. Cell cycle analysis revealed that TRPV3 activation increased the percentage of cells in G2/M phase; consistently, Western blot also revealed a concomitant increase in the expression of inactive form of cyclin‐dependent kinase 1, suggesting that TRPV3 activation arrested mESCs at G2/M phase. TRPV3 activation did not alter the expression of pluripotency markers Oct‐4, Klf4 and c‐Myc, suggesting that the pluripotency was preserved. Our study is the first study to show the presence of TRPV3 at ER. Our study also reveals the novel role of TRPV3 in controlling the cell cycle and preserving the pluripotency of ESCs. J. Cell. Physiol. 231: 403–413, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25086   open full text
• Potential Role of Glycogen Synthase Kinase‐3β in Regulation of Myocardin Activity in Human Vascular Smooth Muscle Cells.
  Yi‐Xia Zhou, Zhan Shi, Pavneet Singh, Hao Yin, Yan‐ni Yu, Long Li, Michael P. Walsh, Yu Gui, Xi‐Long Zheng.
  Journal of Cellular Physiology. October 22, 2015
  Glycogen synthase kinase (GSK)‐3β, a serine/threonine kinase with an inhibitory role in glycogen synthesis in hepatocytes and skeletal muscle, is also expressed in cardiac and smooth muscles. Inhibition of GSK‐3β results in cardiac hypertrophy through reducing phosphorylation and increasing transcriptional activity of myocardin, a transcriptional co‐activator for serum response factor. Myocardin plays critical roles in differentiation of smooth muscle cells (SMCs). This study, therefore, aimed to examine whether and how inhibition of GSK‐3β regulates myocardin activity in human vascular SMCs. Treatment of SMCs with the GSK‐3β inhibitors AR‐A014418 and TWS 119 significantly reduced endogenous myocardin activity, as indicated by lower expression of myocardin target genes (and gene products), CNN1 (calponin), TAGLN1 (SM22), and ACTA2 (SM α‐actin). In human SMCs overexpressing myocardin through the T‐REx system, treatment with either GSK‐3β inhibitor also inhibited the expression of CNN1, TAGLN1, and ACTA2. These effects of GSK‐3β inhibitors were mimicked by transfection with GSK‐3β siRNA. Notably, both AR‐A014418 and TWS 119 decreased the serine/threonine phosphorylation of myocardin. The chromatin immunoprecipitation assay showed that AR‐A014418 treatment reduced myocardin occupancy of the promoter of the myocardin target gene ACTA2. Overexpression of a dominant‐negative GSK‐3β mutant in myocardin‐overexpressing SMCs reduced the expression of calponin, SM22, and SM α‐actin. As expected, overexpression of constitutively active or wild‐type GSK‐3β in SMCs without myocardin overexpression increased expression of these proteins. In summary, our results indicate that inhibition of GSK‐3β reduces myocardin transcriptional activity, suggesting a role for GSK‐3β in myocardin transcriptional activity and smooth muscle differentiation. J. Cell. Physiol. 231: 393–402, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25084   open full text
• Alterations in Skeletal Muscle Oxidative Phenotype in Mice Exposed to 3 Weeks of Normobaric Hypoxia.
  Ilse G.M. Slot, Annemie M.W.J. Schols, Chiel C. de Theije, Frank J.M. Snepvangers, Harry R. Gosker.
  Journal of Cellular Physiology. October 22, 2015
  Skeletal muscle of patients with chronic respiratory failure is prone to loss of muscle mass and oxidative phenotype. Tissue hypoxia has been associated with cachexia and emphysema in humans. Experimental research on the role of hypoxia in loss of muscle oxidative phenotype, however, has yielded inconsistent results. Animal studies are frequently performed in young animals, which may hinder translation to generally older aged patients. Therefore, in this study, we tested the hypothesis that hypoxia induces loss of skeletal muscle oxidative phenotype in a model of aged (52 weeks) mice exposed to 3 weeks of hypoxia. Additional groups of young (4 weeks) and adult (12 weeks) mice were included to examine age effects. To verify hypoxia‐induced cachexia, fat pad and muscle weights as well as muscle fiber cross‐sectional areas were determined. Muscle oxidative phenotype was assessed by expression and activity of markers of mitochondrial metabolism and fiber‐type distribution. A profound loss of muscle and fat was indeed accompanied by a slightly lower expression of markers of muscle oxidative capacity in the aged hypoxic mice. In contrast, hypoxia‐associated changes of fiber‐type composition were more prominent in the young mice. The differential response of the muscle of young, adult, and aged mice to hypoxia suggests that age matters and that the aged mouse is a better model for translation of findings to elderly patients with chronic respiratory disease. Furthermore, the findings warrant further mechanistic research into putative accelerating effects of hypoxia‐induced loss of oxidative phenotype on the cachexia process in chronic respiratory disease. J. Cell. Physiol. 231: 377–392, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25083   open full text
• Platelet‐Derived CCL5 Regulates CXC Chemokine Formation and Neutrophil Recruitment in Acute Experimental Colitis.
  Changhui Yu, Songen Zhang, Yongzhi Wang, Su Zhang, Lingtao Luo, Henrik Thorlacius.
  Journal of Cellular Physiology. October 22, 2015
  Accumulating data suggest that platelets not only regulate thrombosis and haemostasis but also inflammatory processes. Platelets contain numerous potent pro‐inflammatory compounds, including the chemokines CCL5 and CXCL4, although their role in acute colitis remains elusive. The aim of this study is to examine the role of platelets and platelet‐derived chemokines in acute colitis. Acute colitis is induced in female Balb/c mice by administration of 5% dextran sodium sulfate (DSS) for 5 days. Animals receive a platelet‐depleting, anti‐CCL5, anti‐CXCL4, or a control antibody prior to DSS challenge. Colonic tissue is collected for quantification of myeloperoxidase (MPO) activity, CXCL5, CXCL2, interleukin‐6 (IL‐6), and CCL5 levels as well as morphological analyses. Platelet depletion reduce tissue damage and clinical disease activity index in DSS‐exposed animals. Platelet depletion not only reduces levels of CXCL2 and CXCL5 but also levels of CCL5 in the inflamed colon. Immunoneutralization of CCL5 but not CXCL4 reduces tissue damage, CXC chemokine expression, and neutrophil recruitment in DSS‐treated animals. These findings show that platelets play a key role in acute colitis by regulating CXC chemokine generation, neutrophil infiltration, and tissue damage in the colon. Moreover, our results suggest that platelet‐derived CCL5 is an important link between platelet activation and neutrophil recruitment in acute colitis. J. Cell. Physiol. 231: 370–376, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25081   open full text
• Critical Role of AMPK/FoxO3A Axis in Globular Adiponectin‐Induced Cell Cycle Arrest and Apoptosis in Cancer Cells.
  Anup Shrestha, Saroj Nepal, Mi Jin Kim, Jae Hoon Chang, Sang‐Hyun Kim, Gil‐Saeng Jeong, Chul‐Ho Jeong, Gyu Hwan Park, Sunghee Jung, Jaecheong Lim, Eunha Cho, Soyoung Lee, Pil‐Hoon Park.
  Journal of Cellular Physiology. October 22, 2015
  Adiponectin predominantly secreted from adipose tissue has exhibited potent anti‐proliferative properties in cancer cells via modulating cell cycle and apoptosis. FoxO3A, a Forkhead box O member of the transcription factor, plays a critical role in modulating expression of genes involved in cell death and/or survival. In this study, we investigated the role of FoxO3A signaling in anti‐cancer activities of adiponectin. Herein, we have shown that treatment with globular adiponectin (gAcrp) increases p27 but decreases cyclinD1 expression in human hepatoma (HepG2) and breast (MCF‐7) cancer cells. Gene ablation of FoxO3A prevented gAcrp‐induced increase in p27 and decreased in cyclin D1 expression, and further ameliorated cell cycle arrest by gAcrp, indicating a critical role of FoxO3A in gAcrp‐induced cell cycle arrest of cancer cells. Moreover, treatment with gAcrp also induced caspase‐3/7 activation and increased Fas ligand (FasL) expression in both HepG2 and MCF‐7 cells. Transfection with FoxO3A siRNA inhibited gAcrp‐induced caspase‐3/7 activation and FasL expression, suggesting that FoxO3A signaling also plays an important role in gAcrp‐induced apoptosis of cancer cells. We also found that gene silencing of AMPK prevented gAcrp‐induced nuclear translocation of FoxO3A in HepG2 and MCF‐7 cells. In addition, suppression of AMPK also blocked gAcrp‐induced cell cycle arrest and further attenuated gAcrp‐induced caspase‐3/7 activation, indicating that AMPK signaling plays a pivotal role in both gAcrp‐induced cell cycle arrest and apoptosis via acting as an upstream signaling of FoxO3A. Taken together, our findings demonstrated that AMPK/FoxO3A axis plays a cardinal role in anti‐proliferative effect of adiponectin in cancer cells. J. Cell. Physiol. 231: 357–369, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25080   open full text
• Genetic and Functional Analysis of Polymorphisms in the Human Dopamine Receptor and Transporter Genes in Small Cell Lung Cancer.
  Emanuela Cherubini, Arianna Di Napoli, Alessia Noto, Giorgia Amira Osman, Maria Cristina Esposito, Salvatore Mariotta, Rossella Sellitri, Luigi Ruco, Giuseppe Cardillo, Gennaro Ciliberto, Rita Mancini, Alberto Ricci.
  Journal of Cellular Physiology. October 22, 2015
  The regulatory role of dopamine (DA) in endocrine, cardiovascular and renal functions has been extensively studied and used for clinical purposes. More recently DA has been indicated as a regulatory molecule for immune cells and malignant cell proliferation. We assessed the expression and the functional role DA, DA receptors, and transporters in primary small cell lung cancer (SCLC). By HPLC DA plasma levels were more elevated in SCLC patients in comparison with NSCLC patients and healthy controls. SCLC cell expressed DA D1‐ and D2‐like receptors and membrane and vesicular transporters at protein and mRNA levels. We also investigated the effects of independent D1‐ or D2‐like receptor stimulation on SCLC cell cultures. DA D1 receptor agonist SKF38393 induced the increase of cAMP levels and DARPP‐32 protein expression without affecting SCLC growth rate. Cell treatment with the DA D1 receptor antagonist SCH23390 inhibited SKF38393 effects. In contrast, the DA D2 receptor agonist quinpirole (10 μM) counteracted, in a dose and time dependent way, SCLC cell proliferation, it did not affect cAMP levels and decreased phosphorylated AKT that was induced by DA D2 receptor antagonist sulpiride. However, in only one SCLC line, stimulation of DA D2 receptor failed to inhibit cell proliferation in vitro. This effect was associated to the existence of rs6275 and rs6277 polymorphisms in the D2 gene. These results gave more insight into DA control of lung cancer cell behavior and suggested the existence of different SCLC phenotypes. J. Cell. Physiol. 231: 345–356, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25079   open full text
• p14ARF Prevents Proliferation of Aneuploid Cells by Inducing p53‐Dependent Apoptosis.
  Lorena Veneziano, Viviana Barra, Laura Lentini, Sergio Spatafora, Aldo Di Leonardo.
  Journal of Cellular Physiology. October 22, 2015
  Weakening the Spindle Assembly Checkpoint by reduced expression of its components induces chromosome instability and aneuploidy that are hallmarks of cancer cells. The tumor suppressor p14ARF is overexpressed in response to oncogenic stimuli to stabilize p53 halting cell progression. Previously, we found that lack or reduced expression of p14ARF is involved in the maintenance of aneuploid cells in primary human cells, suggesting that it could be part of a pathway controlling their proliferation. To investigate this aspect further, p14ARF was ectopically expressed in HCT116 cells after depletion of the Spindle Assembly Checkpoint MAD2 protein that was used as a trigger for aneuploidy. p14ARF Re‐expression reduced the number of aneuploid cells in MAD2 post‐transcriptionally silenced cells. Also aberrant mitoses, frequently displayed in MAD2‐depleted cells, were decreased when p14ARF was expressed at the same time. In addition, p14ARF ectopic expression in MAD2‐depleted cells induced apoptosis associated with increased p53 protein levels. Conversely, p14ARF ectopic expression did not induce apoptosis in HCT116 p53KO cells. Collectively, our results suggest that the tumor suppressor p14ARF may have an important role in counteracting proliferation of aneuploid cells by activating p53‐dependent apoptosis. J. Cell. Physiol. 231: 336–344, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.24976   open full text
• Ca2+ Signalling in Endothelial Progenitor Cells: Friend or Foe?
  Francesco Moccia, Germano Guerra.
  Journal of Cellular Physiology. October 22, 2015
  Endothelial progenitor cells (EPCs) are mobilized either from the bone marrow and/or the arterial to replace dysfunctional endothelial cells and rescue blood perfusion in ischemic tissues. In addition, they may contribute to the angiogenic switch, thereby sustaining tumour growth and metastatization. Understanding the molecular mechanisms utilized by vascular endothelial growth factor (VEGF) to stimulate EPCs might unveil novel targets to enhance their clinical outcome in regenerative medicine and to adverse tumour vascularisation. VEGF stimulates peripheral blood‐derived EPCs to undergo repetitive Ca2+ oscillations shaped by the interaction between inositol‐1,4,5‐trisphosphate (InsP3)‐dependent Ca2+ release and store‐operated Ca2+ entry (SOCE). However, the Ca2+ machinery underlying VEGF‐induced Ca2+ spikes changes in umbilical cord blood‐derived EPCs, which require TRPC3‐mediated Ca2+ entry to trigger the interplay between InsP3 and SOCE. Surprisingly, VEGF fails to elicit pro‐angiogenic Ca2+ signals when EPCs derive from renal cellular carcinoma patients, thus questioning the suitability of VEGFR‐2 as a target for anti‐angiogenic treatments in these individuals. The lack of response to VEGF is likely due to the dramatic rearrangement of the Ca2+ toolkit occurring in RCC‐derived EPCs. Finally, primary myelofibrosis‐derived EPCs display a further pattern of reorganization of the Ca2+ machinery and proliferate independently of SOCE. Thus, the Ca2+ machinery in human ECFCs is extremely plastic and may change depending on the physio‐pathological background of the donor. As a consequence, the Ca2+ toolkit could properly be used to enhance the regenerative outcome of cell‐based therapy or adverse tumor vascularisation. J. Cell. Physiol. 231: 314–327, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25126   open full text
• MicroRNAs in Control of Plant Development.
  Chao Li, Baohong Zhang.
  Journal of Cellular Physiology. October 22, 2015
  In the long evolutionary history, plant has evolved elaborate regulatory network to control functional gene expression for surviving and thriving, such as transcription factor‐regulated transcriptional programming. However, plenty of evidences from the past decade studies demonstrate that the 21–24 nucleotides small RNA molecules, majorly microRNAs (miRNAs) play dominant roles in post‐transcriptional gene regulation through base pairing with their complementary mRNA targets, especially prefer to target transcription factors in plants. Here, we review current progresses on miRNA‐controlled plant development, from miRNA biogenesis dysregulation‐caused pleiotropic developmental defects to specific developmental processes, such as SAM regulation, leaf and root system regulation, and plant floral transition. We also summarize some miRNAs that are experimentally proved to greatly affect crop plant productivity and quality. In addition, recent reports show that a single miRNA usually displays multiple regulatory roles, such as organ development, phase transition, and stresses responses. Thus, we infer that miRNA may act as a node molecule to coordinate the balance between plant development and environmental clues, which may shed the light on finding key regulator or regulatory pathway for uncovering the mysterious molecular network. J. Cell. Physiol. 231: 303–313, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25125   open full text
• Cellular and Molecular Mechanisms of Phenotypic Switch in Gastrointestinal Smooth Muscle.
  Annunziata Scirocco, Paola Matarrese, Marilia Carabotti, Barbara Ascione, Walter Malorni, Carola Severi.
  Journal of Cellular Physiology. October 22, 2015
  As a general rule, smooth muscle cells (SMC) are able to switch from a contractile phenotype to a less mature synthetic phenotype. This switch is accompanied by a loss of differentiation with decreased expression of contractile markers, increased proliferation as well as the synthesis and the release of several signaling molecules such as pro‐inflammatory cytokines, chemotaxis‐associated molecules, and growth factors. This SMC phenotypic plasticity has extensively been investigated in vascular diseases, but interest is also emerging in the field of gastroenterology. It has in fact been postulated that altered microenvironmental conditions, including the composition of microbiota, could trigger the remodeling of the enteric SMC, with phenotype changes and consequent alterations of contraction and impairment of gut motility. Several molecular actors participate in this phenotype remodeling. These include extracellular molecules such as cytokines and extracellular matrix proteins, as well as intracellular proteins, for example, transcription factors. Epigenetic control mechanisms and miRNA have also been suggested to participate. In this review key roles and actors of smooth muscle phenotypic switch, mainly in GI tissue, are described and discussed in the light of literature data available so far. J. Cell. Physiol. 231: 295–302, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25105   open full text
• Endoplasmic Reticulum Stress Interacts With Inflammation in Human Diseases.
  Stewart Siyan Cao, Katherine L. Luo, Lynn Shi.
  Journal of Cellular Physiology. October 22, 2015
  The endoplasmic reticulum (ER) is a critical organelle for normal cell function and homeostasis. Disturbance in the protein folding process in the ER, termed ER stress, leads to the activation of unfolded protein response (UPR) that encompasses a complex network of intracellular signaling pathways. The UPR can either restore ER homeostasis or activate pro‐apoptotic pathways depending on the type of insults, intensity and duration of the stress, and cell types. ER stress and the UPR have recently been linked to inflammation in a variety of human pathologies including autoimmune, infectious, neurodegenerative, and metabolic disorders. In the cell, ER stress and inflammatory signaling share extensive regulators and effectors in a broad spectrum of biological processes. In spite of different etiologies, the two signaling pathways have been shown to form a vicious cycle in exacerbating cellular dysfunction and causing apoptosis in many cells and tissues. However, the interaction between ER stress and inflammation in many of these diseases remains poorly understood. Further understanding of the biochemistry, cell biology, and physiology may enable the development of novel therapies that spontaneously target these pathogenic pathways. J. Cell. Physiol. 231: 288–294, 2016. © 2015 Wiley Periodicals, Inc.

  October 22, 2015   doi: 10.1002/jcp.25098   open full text
• Appl1 and Appl2 are Expendable for Mouse Development But Are Essential for HGF‐Induced Akt Activation and Migration in Mouse Embryonic Fibroblasts.
  Yinfei Tan, Xiaoban Xin, Francis J. Coffey, David L. Wiest, Lily Q. Dong, Joseph R. Testa.
  Journal of Cellular Physiology. October 21, 2015
  Although Appl1 and Appl2 have been implicated in multiple cellular activities, we and others have found that Appl1 is dispensable for mouse embryonic development, suggesting that Appl2 can substitute for Appl1 during development. To address this possibility, we generated conditionally targeted Appl2 mice. We found that ubiquitous Appl2 knockout (Appl2−/−) mice, much like Appl1−/− mice, are viable and grow normally to adulthood. Intriguingly, when Appl1−/− mice were crossed with Appl2−/− mice, we found that homozygous Appl1;Appl2 double knockout (DKO) animals are also viable and grossly normal with regard to reproductive potential and postnatal growth. Appl2‐null and DKO mice were found to exhibit altered red blood cell physiology, with erythrocytes from these mice generally being larger and having a more irregular shape than erythrocytes from wild type mice. Although Appl1/2 proteins have been previously shown to have a very strong interaction with phosphatidylinositol‐3 kinase (Pi3k) in thymic T cells, Pi3k‐Akt signaling and cellular differentiation was unaltered in thymocytes from Appl1;Appl2 (DKO) mice. However, Appl1/2‐null mouse embryonic fibroblasts exhibited defects in HGF‐induced Akt activation, migration, and invasion. Taken together, these data suggest that Appl1 and Appl2 are required for robust HGF cell signaling but are dispensable for embryonic development and reproduction. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  October 21, 2015   doi: 10.1002/jcp.25211   open full text
• Calcineurin‐NFAT Signaling Controls Somatic Cell Reprogramming in a Stage‐Dependent Manner.
  Ming Sun, Bing Liao, Yu Tao, Hao Chen, Feng Xiao, Junjie Gu, Shaorong Gao, Ying Jin.
  Journal of Cellular Physiology. October 20, 2015
  Calcineurin‐NFAT signaling is critical for early lineage specification of mouse embryonic stem cells and early embryos. However, its roles in somatic cell reprogramming remain unknown. Here, we report that calcineurin‐NFAT signaling has a dynamic activity and plays diverse roles at different stages of reprogramming. At the early stage, calcineurin‐NFAT signaling is transiently activated and its activation is required for successful reprogramming. However, at the late stage of reprogramming, activation of calcineurin‐NFAT signaling becomes a barrier for reprogramming and its inactivation is critical for successful induction of pluripotency. Mechanistically, calcineurin‐NFAT signaling contributes to the reprogramming through regulating multiple early events during reprogramming, including mesenchymal to epithelial transition (MET), cell adhesion and emergence of SSEA1+ intermediate cells. Collectively, this study reveals for the first time the important roles of calcineurin‐NFAT signaling during somatic cell reprogramming and provides new insights into the molecular regulation of reprogramming. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  October 20, 2015   doi: 10.1002/jcp.25212   open full text
• BMP‐2 Enhances Lgr4 Gene Expression in Osteoblastic Cells.
  Chantida Pawaputanon Na Mahasarakham, Yoichi Ezura, Makiri Kawasaki, Arayal Smriti, Shuichi Moriya, Takayuki Yamada, Yayoi Izu, Akira Nifuji, Katsuhiko Nishimori, Yuichi Izumi, Masaki Noda.
  Journal of Cellular Physiology. October 19, 2015
  Osteoporosis is one of the most prevalent diseases and the number of patients suffering from this disease is soaring due to the increase in the aged population in the world. The severity of bone loss in osteoporosis is based on the levels of impairment in the balance between bone formation and bone resorption, two arms of the bone metabolism, and bone remodeling. However, determination of bone formation levels is under many layers of control that are as yet fully defined. Bone morphogenetic protein (BMP) plays a key role in regulation of bone formation while its downstream targets are still incompletely understood. Lgr4 gene encodes an orphan receptor and has been identified as a genetic determinant for bone mass in osteoporotic patients. Here, we examine the effects of BMP on the expression of Lgr4 in osteoblastic cells. Lgr4 gene is expressed in an osteoblastic cell line, MC3T3E1 in a time dependent manner during the culture. BMP treatment enhances Lgr4 mRNA expression at least in part via transcriptional event. When Lgr4 mRNA is knocked down, the levels of BMP‐induced increase in alkaline phosphatase (Alp) activity and Alp mRNA are suppressed. BMP enhancement of Lgr4 gene expression is suppressed by FGF and reversed by dexamethasone. BMP also enhances Lgr4 expression in primary cultures of calvarial osteoblasts. These data indicate that Lgr4 gene is regulated by BMP and is required for BMP effects on osteoblastic differentiation. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  October 19, 2015   doi: 10.1002/jcp.25180   open full text
• The Nox1/4 Dual Inhibitor GKT137831 or Nox4 Knockdown Inhibits Angiotensin‐II‐Induced Adult Mouse Cardiac Fibroblast Proliferation and Migration. AT1 Physically Associates With Nox4.
  Naveen K. Somanna, Anthony J. Valente, Maike Krenz, William P. Fay, Patrice Delafontaine, Bysani Chandrasekar.
  Journal of Cellular Physiology. October 19, 2015
  Both oxidative stress and inflammation contribute to chronic hypertension‐induced myocardial fibrosis and adverse cardiac remodeling. Here we investigated whether angiotensin (Ang)‐II‐induced fibroblast proliferation and migration are NADPH oxidase (Nox) 4/ROS and IL‐18 dependent. Our results show that the potent induction of mouse cardiac fibroblast (CF) proliferation and migration by Ang‐II is markedly attenuated by Nox4 knockdown and the Nox inhibitor DPI. Further, Nox4 knockdown and DPI pre‐treatment attenuated Ang‐II‐induced IL‐18, IL‐18Rα and collagen expression, and MMP9 and LOX activation. While neutralization of IL‐18 blunted Ang‐II‐induced CF proliferation and migration, knockdown of MMP9 attenuated CF migration. The antioxidant NAC and the cell‐permeable SOD mimetics Tempol, MnTBAP, and MnTMPyP attenuated oxidative stress and inhibited CF proliferation and migration. The Nox1/Nox4 dual inhibitor GKT137831 also blunted Ang‐II‐induced H2O2 production and CF proliferation and migration. Further, AT1 bound Nox4, and Ang‐II enhanced their physical association. Notably, GKT137831 attnuated the AT1/Nox4 interaction. These results indicate that Ang‐II induces CF proliferation and migration in part via Nox4/ROS‐dependent IL‐18 induction and MMP9 activation, and may involve AT1/Nox4 physical association. Thus, either (i) neutralizing IL‐18, (ii) blocking AT1/Nox4 interaction or (iii) use of the Nox1/Nox4 inhibitor GKT137831 may have therapeutic potential in chronic hypertension‐induced adverse cardiac remodeling. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  October 19, 2015   doi: 10.1002/jcp.25210   open full text
• Definitive Hematopoietic Multipotent Progenitor Cells Are Transiently Generated From Hemogenic Endothelial Cells in Human Pluripotent Stem Cells.
  Hao Bai, Yanfeng Liu, Yinliang Xie, Dixie L. Hoyle, Robert A. Brodsky, Linzhao Cheng, Tao Cheng, Zack Z. Wang.
  Journal of Cellular Physiology. October 14, 2015
  Generation of fully functional hematopoietic multipotent progenitor (MPP) cells from human pluripotent stem cells (hPSCs) has a great therapeutic potential to provide an unlimited cell source for treatment of hematological disorders. We previously demonstrated that CD34+CD31+CD144+ population derived from hPSCs contain hemato‐endothelial progenitors (HEPs) that give rise to hematopoietic and endothelial cells. Here, we report a differentiation system to generate definitive hematopoietic MPP cells from HEPs via endothelial monolayer. In the presence of angiogenic factors, HEPs formed an endothelial monolayer, from which hematopoietic clusters emerged through the process of endothelial‐to‐hematopoietic transition (EHT). EHT was significantly enhanced by hematopoietic growth factors. The definitive MPP cells generated from endothelial monolayer were capable of forming multilineage hematopoietic colonies, giving rise to T lymphoid cells, and differentiating into enucleated erythrocytes. Emergence of hematopoietic cells from endothelial monolayer occurred transiently. Hematopoietic potential was lost during prolonged culture of HEPs in endothelial growth conditions. Our study demonstrated that CD34+CD31+CD144+ HEPs gave rise to hematopoietic MPP cells via hemogenic endothelial cells that exist transiently. The established differentiation system provides a platform for future investigation of regulatory factors involved in de novo generation of hematopoietic MPP cells and their applications in transplantation. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  October 14, 2015   doi: 10.1002/jcp.25199   open full text
• Cathepsin K Deficiency Suppresses Disuse‐Induced Bone Loss.
  Shuichi Moriya, Yayoi Izu, Smriti Arayal, Makiri Kawasaki, Koki Hata, Chantida Pawaputanon Na Mahasarakhahm, Yuichi Izumi, Paul Saftig, Kazuo Kaneko, Masaki Noda, Yoichi Ezura.
  Journal of Cellular Physiology. October 13, 2015
  Unloading induces bone loss and causes disuse osteoporosis. However, the mechanism underlying disuse osteoporosis is still incompletely understood. Here we examined the effects of cathepsin K (CatK) deficiency on disuse osteoporosis induced by using sciatic neurectomy (Nx) model. After four weeks of surgery, Cat‐K KO and WT mice were sacrificed and subjected to analyses. For cancellous bone rich region, Nx reduced the bone mineral density (BMD) compared to the BMD in the sham operated side in wild type mice. In contrast, CatK deficiency suppressed such Nx‐induced reduction of BMD in cancellous bone. Nx also reduced BMD in the mid shaft cortical bone compared to the BMD in the corresponding region on the sham operated side in wild type mice. In contrast, CatK deficiency suppressed such Nx‐induced reduction of BMD in the mid shaft cortical bone. Bone volume (BV/TV) was reduced by Nx in WT mice. In contrast, Cat‐K deficiency suppressed such reduction in bone volume. Interestingly, CatK deficiency suppressed osteoclast number and osteoclast surface in the Nx side compared to sham side. When bone marrow cells obtained from Nx side femur of CatK‐KO mice were cultured, the levels of the calcified area in culture were increased. Further examination of gene expression indicated that Nx suppressed the expression of genes encoding osteoblast‐phenotype‐related molecules such as Runx2 and alkaline phosphatase in WT mice. In contrast, CatK deficiency suppressed such reduction. These data indicate that CatK is involved in the disuse‐induced bone mass reduction. This article is protected by copyright. All rights reserved

  October 13, 2015   doi: 10.1002/jcp.25214   open full text
• Lipidomics of Mesenchymal Stromal Cells: Understanding the Adaptation of Phospholipid Profile in Response to Pro‐Inflammatory Cytokines.
  Ana Margarida Campos, Elisabete Maciel, Ana S. P. Moreira, Bebiana Sousa, Tânia Melo, Pedro Domingues, Liliana Curado, Brígida Antunes, M. Rosário M. Domingues, Francisco Santos.
  Journal of Cellular Physiology. October 08, 2015
  Mesenchymal stromal cells (MSCs) present anti‐inflammatory properties and are being used with great success as treatment for inflammatory and autoimmune diseases. In clinical applications MSCs are subjected to a strong pro‐inflammatory environment, essential to their immunosuppressive action. Despite the wide clinical use of these cells, how MSCs exert their effect remains unclear. Several lipids are known to be involved in cell's signaling and modulation of cellular functions. The aim of this paper is to examine the variation in lipid profile of MSCs under pro‐inflammatory environment, induced by the presence of tumor necrosis factor alpha (TNF‐α) and interferon gamma (IFN‐γ), using the most modern lipidomic approach. Major changes in lipid molecular profile of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), lysoPC (LPC), and sphingomyelin (SM) classes were found. No changes were observed in the phosphatidylinositol (PI) profile. The levels of PC species with shorter fatty acids (FAs), mainly C16:0, decreased under pro‐inflammatory stimuli. The level of PC(40:6) also decreased, which may be correlated with enhanced levels of LPC(18:0), which is known to be an anti‐inflammatory LPC, observed in MSCs subjected to TNF‐α and IFN‐γ. Simultaneously, the relative amounts of PC(36:1) and PC(38:4) increased. TNF‐α and IFN‐γ also enhanced the levels of PE(40:6) and decreased the levels of PE(O‐38:6). Higher expression of PS(36:1) and SM(34:0) along with a decrease in PS(38:6) levels were observed. These results indicate that lipid metabolism and signaling are modulated during MSCs activation, which suggests that lipids may be involved in MSCs functional and anti‐inflammatory activities. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  October 08, 2015   doi: 10.1002/jcp.25191   open full text
• Smooth Muscle Cell Differentiation: Model Systems, Regulatory Mechanisms, and Vascular Diseases.
  Ning Shi, Shi‐You Chen.
  Journal of Cellular Physiology. October 08, 2015
  Smooth muscle cell (SMC) differentiation is an important process during vascular development. The highly differentiated mature SMCs play critical roles in maintaining structural and functional integrity of blood vessels. However, SMCs are not terminally differentiated, and their phenotype can be modulated between contractile and proliferative states in response to various environmental conditions. Alterations in SMC phenotype contribute to a number of major cardiovascular diseases such as atherosclerosis, hypertension and restenosis following angioplasty, etc. The goal of this review is to provide a brief overview of the recent advancements in our understanding of SMC differentiation and the development of in vitro SMC differentiation models, with a particular emphasis on examination of molecular mechanisms involved in the regulation of SMC differentiation and phenotypic modulation. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  October 08, 2015   doi: 10.1002/jcp.25208   open full text
• Involvement of Notch‐1 in Resistance to Regorafenib in Colon Cancer Cells.
  Giovanna Mirone, Stefania Perna, Arvind Shukla, Gabriella Marfe.
  Journal of Cellular Physiology. October 08, 2015
  Regorafenib, an oral small‐molecule multi kinase inhibitor, is able to block Vascular Endothelial Growth Factor Receptors (VEGFR‐1, 2, and 3), Platelet‐Derived Growth Factor Receptors (PDGF), Fibroblast Growth Factor (FGF) receptor 1, Raf, TIE‐2, and the kinases KIT, RET, and BRAF. Different studies have displayed its antitumor activity in several cancer models (both in vitro and in vivo), particularly in colorectal and gastrointestinal stromal cancers. The mechanism of resistance to regorafenib is largely unknown. In our investigation, we have generated regorafenib‐resistant SW480 cells (Reg‐R‐SW480 cells) by culturing such cells with increasing concentration of regorafenib. Examination of intracellular signaling found that Akt signaling was activated in Reg‐R‐SW480 cells but not in wild–type SW480 cells, after regorafenib treatment as measured by Western Blot. The Notch pathway is a fundamental signaling system in the development and homeostasis of tissues since it regulates different cellular process such as proliferation, differentiation, and apoptosis and it can be a potential driver of resistance to a wide array of targeted therapies. In this study, we found that Notch‐1 was significantly up‐regulated in resistant tumor cells as well as HES1 and HEY. Additionally, inhibition of Notch‐1 in resistant cells partially restored sensitivity to regorafenib treatment in vitro. Collectively, these data suggest a key role of Notch‐1 in mediating the resistant effects of regorafenib in colorectal cancer cells, and also provide a rationale to improve the therapeutic efficacy of regorafenib. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  October 08, 2015   doi: 10.1002/jcp.25206   open full text
• MDA‐9/Syntenin Control.
  Julie V. Philley, Anbarasu Kannan, Santanu Dasgupta.
  Journal of Cellular Physiology. October 07, 2015
  MDA‐9/Syntenin is a small PDZ domain containing scaffolding protein with diverse array of functions regulating membrane trafficking, cell adhesion, neural, and synaptic development, ubiquitination, and exosome biogenesis. An appreciable number of studies also established a pivotal role of MDA‐9/Syntenin in cancer development and progression. In this review, we will discuss the dynamic role of MDA‐9/Syntenin in regulating normal and abnormal fate of various cellular processes. J. Cell. Physiol. 9999: 1–6, 2015. © 2015 Wiley Periodicals, Inc.

  October 07, 2015   doi: 10.1002/jcp.25136   open full text
• The IP3R Binding Protein Released with Inositol 1,4,5‐Trisphosphate Is Expressed in Rodent Reproductive Tissue and Spermatozoa.
  Heike Borth, Nele Weber, Dorke Meyer, Andrea Wartenberg, Elisabeth Arlt, Susanna Zierler, Andreas Breit, Gunther Wennemuth, Thomas Gudermann, Ingrid Boekhoff.
  Journal of Cellular Physiology. October 06, 2015
  Besides its capacity to inhibit the 1,4,5‐trisphosphate (IP3) receptor, the regulatory protein IRBIT (IP3 receptor‐binding protein released with IP3) is also able to control the activity of numerous ion channels and electrolyte transporters and thereby creates an optimal electrolyte composition of various biological fluids. Since a reliable execution of spermatogenesis and sperm maturation critically depends on the establishment of an adequate microenvironment, the expression of IRBIT in male reproductive tissue was examined using immunohistochemical approaches combined with biochemical fractionation methods. The present study documents that IRBIT is expressed in Leydig and Sertoli cells. In addition, pronounced IRBIT expression was detected in sperm precursors during early stages of spermatogenesis as well as in spermatozoa. Analyzing tissue sections of rodent epididymides, IRBIT was found to co‐localize with the proton pumping V‐ATPase and the cystic fibrosis transmembrane conductance regulator (CFTR) at the apical surface of narrow and clear cells. A similar co‐localization of IRBIT with CFTR was also observed for Sertoli cells and developing germ cells. Remarkably, assaying caudal sperm in immunogold electron microscopy, IRBIT was found to localize to the acrosomal cap and the flagellum as well as to the sperm nucleus; moreover, a prominent oligomerization was observed for spermatozoa. The pronounced occurrence of IRBIT in the male reproductive system and mature spermatozoa indicates a potential role for IRBIT in establishing the essential luminal environment for a faithful execution of spermatogenesis and epididymal sperm maturation, and suggest a participation of IRBIT during maturations steps after ejaculation and/or the final fertilization process. This article is protected by copyright. All rights reserved

  October 06, 2015   doi: 10.1002/jcp.25209   open full text
• Epigenetic Modulation in Periodontitis: Interaction of Adiponectin and JMJD3‐IRF4 Axis in Macrophages.
  Dongying Xuan, Qianqian Han, Qisheng Tu, Lan Zhang, Liming Yu, Dana Murry, Tianchi Tu, Yin Tang, Jane Lian, Gary S. Stein, Paloma Valverde, Jincai Zhang, Jake Chen.
  Journal of Cellular Physiology. October 06, 2015
  Emerging evidence suggests an important role for epigenetic mechanisms in modulating signals during macrophage polarization and inflammation. JMJD3, a JmjC family histone demethylase necessary for M2 polarization is also required for effective induction of multiple M1 genes by lipopolysaccharide (LPS). However, the effects of JMJD3 to inflammation in the context of obesity remains unknown. To address this deficiency, we firstly examined the expression of JMJD3 in macrophage isolated from bone marrow and adipose tissue of diet induced obesity (DIO) mice. The results indicated that JMJD3 was down‐regulated in obesity. Adiponectin (APN), a factor secreted by adipose tissue which is down‐regulated in obesity, functions to switch macrophage polarization from M1 to M2, thereby attenuating chronic inflammation. Intriguingly, our results indicated that APN contributed to JMJD3 up‐regulation, reduced macrophage infiltration in obese adipose tissue, and abolished the up‐regulation of JMJD3 in peritoneal macrophages isolated from DIO mice when challenged with Porphyromonas gingivalis LPS (pg.lps). To elucidate the interaction of APN and JMJD3 involved in macrophage transformation in the context of inflammation, we designed the loss and gain‐function experiments of APN in vivo with APN−/− mice with experimental periodontitis and in vitro with macrophage isolated from APN−/− mice. For the first time, we found that APN can help to reduce periodontitis‐related bone loss, modulate JMJD3 and IRF4 expression, and macrophage infiltration. Therefore, it can be inferred that APN may contribute to anti‐inflammation macrophage polarization by regulating JMJD3 expression, which provides a basis for macrophage‐centered epigenetic therapeutic strategies. J. Cell. Physiol. 9999: 1–7, 2015. © 2015 Wiley Periodicals, Inc.

  October 06, 2015   doi: 10.1002/jcp.25201   open full text
• Reduced dentin matrix protein expression in Camurati‐Engelmann disease transgenic mouse model.
  Angela Gullard, Christina M. Croney, Xiangwei Wu, Olga Mamaeva, Philip Sohn, Xu Cao, Mary MacDougall.
  Journal of Cellular Physiology. October 01, 2015
  Overexpression of transforming growth factor‐beta 1 (TGF‐β1) has been shown to lead to mineralization defects in both the enamel and dentin layers of teeth. A TGFB1 point mutation (H222D), derived from published cases of Camurati‐Engelmann disease (CED), has been shown to constitutively activate TGF‐β1, leading to excess bone matrix production. Although CED has been well documented in clinical case reports, there are no published studies on the effect of CED on the dentition. The objective of this study was to determine the dental manifestations of hyperactivated TGF‐β1 signaling using an established mouse model of CED‐derived TGF‐β1 mutation. Murine dental tissues were studied via radiography, micro‐CT, immunohistochemistry, and qRT‐PCR. Results showed that initial decreased dental mineralized tissue density is resolved Proliferation assays of incisor pulp and alveolar bone cell cultures revealed that cells from transgenic animals displayed a reduced rate of growth compared to alveolar bone cultures from wild‐type mice. TGF‐β family gene expression analysis indicated significant fold changes in the expression of Alpl, Bmp2‐5, Col‐1, ‐2,‐4 and ‐6, Fgf, Mmp, Runx2, Tgfb3, Tfgbr3, and Vdr genes. Assessment of SIBLINGs reveals downregulation of Ibsp, Dmp1, Dspp, Mepe, and Spp1, as well as reduced staining for BMP‐2 and VDR in mesenchymal‐derived pulp tissue in CED animals. Treatment of dental pulp cells with recombinant human TGF‐β1 resulted in increased SIBLING gene expression. Conclusions: Our results provide in vivo evidence suggesting that TFG‐β1 mediates expression of important dentin extracellular matrix components secreted by dental pulp, and when unbalanced, may contribute to abnormal dentin disorders. This article is protected by copyright. All rights reserved

  October 01, 2015   doi: 10.1002/jcp.25207   open full text
• Evidences of Biological Functions of Biliverdin Reductase A in the Bovine Epididymis.
  Olivier D'Amours, Gilles Frenette, Patrick Caron, Clémence Belleannée, Chantal Guillemette, Robert Sullivan.
  Journal of Cellular Physiology. September 30, 2015
  Epididymal sperm binding protein 1 (ELSPBP1) is secreted by the epididymal epithelium via epididymosomes and is specifically transferred to dead spermatozoa during epididymal transit. We identified biliverdin reductase A (BLVRA) as a partner of ELSPBP1 by immunoprecipitation followed by tandem mass spectrometry. Pull down assays showed that these two proteins interact in the presence of zinc ions. The BLVRA enzyme is known to convert biliverdin to bilirubin, both of which possess antioxidant activity. Assessment by real‐time RT‐PCR showed that BLVRA is highly expressed in the caput and the corpus epididymis, but is expressed at lower levels in the testis and the cauda epididymis. It is primarily found in the soluble fraction of the caput epididymal fluid, is barely detectable in the cauda fluid, and is detectable to a lesser extent in the epididymosome fraction of both caput and cauda fluids. Immunocytometry on epididymal sperm showed that BLVRA is found on all sperm recovered from the caput region, whereas it is undetectable on cauda sperm. Biliverdin and bilirubin are found in higher concentrations in the caput epididymal fluid, as measured by mass spectrometry. Lipid peroxidation was limited by 1 μM of biliverdin, but not bilirubin when caput spermatozoa were challenged with 500 μM H2O2. Since immature spermatozoa are a source of reactive oxygen species, BLVRA may be involved in the protection of maturing spermatozoa. It is also plausible that BLVRA is implicated in haemic protein catabolism in the epididymal luminal environment. J. Cell. Physiol. 9999: 1–13, 2015. © 2015 Wiley Periodicals, Inc.

  September 30, 2015   doi: 10.1002/jcp.25200   open full text
• Genes in the GABA Pathway Increase in the Lateral Thalamus of Sprague–Dawley Rats During the Proestrus/Estrus Phase.
  Mikhail Umorin, Crystal Stinson, Larry L. Bellinger, Phillip R. Kramer.
  Journal of Cellular Physiology. September 30, 2015
  Pain can vary over the estrous cycle as a result of changes in estradiol concentration but the mechanism causing this variation is unclear. Because the thalamus is important in pain control, gene expression in the lateral thalamus (ventral posteromedial, ventral posterolateral, reticular thalamic nuclei) was screened at different phases of the estrous cycle. Gene expression changes in Sprague–Dawley rats were further analyzed by real‐time PCR and ELISA and plasma estradiol levels were measured by RIAs at different phases of the estrous cycle. Our results indicated that both the RNA and protein expression of glutamate decarboxylase 1 and 2 (GAD1, GAD2), GABA(A) receptor‐associated protein like 1 (GABARAPL1), and vesicular GABA transporter (VGAT) significantly increased in the lateral thalamus when plasma estradiol levels were elevated. Estradiol levels were elevated during the proestrus and estrus phases of the estrous cycle. Estrogen receptor α (ERα) was observed to be co‐localized in thalamic cells and thalamic infusion of an ERα antagonist significantly reduced GAD1 and VGAT transcript. GAD1, GAD2, GABARAPL1, and VGAT have been shown to effect neuronal responses suggesting that attenuation of pain during the estrous cycle can be dependent, in part, through estradiol induced changes in thalamic gene expression. J. Cell. Physiol. 9999: 1–8, 2015. © 2015 Wiley Periodicals, Inc.

  September 30, 2015   doi: 10.1002/jcp.25198   open full text
• Glucose and Inflammatory Cells Decrease Adiponectin in Epicardial Adipose Tissue Cells: Paracrine Consequences on Vascular Endothelium.
  Ángel Fernández‐Trasancos, Raquel Guerola‐Segura, Beatriz Paradela‐Dobarro, Ezequiel Álvarez, José María García‐Acuña, Ángel Luis Fernández, José Ramón González‐Juanatey, Sonia Eiras.
  Journal of Cellular Physiology. September 29, 2015
  Epicardial adipose tissue (EAT) is a source of energy for heart that expresses the insulin‐sensitizer, anti‐inflammatory and anti‐atherogenic protein, adiponectin. But, in coronary artery disease, adiponectin production declines. Our objective was to determine its regulation by glucose and inflammation in stromal cells from EAT and subcutaneous adipose tissue (SAT) and its paracrine effect on endothelial cells. Stromal cells of EAT and SAT were obtained from patients who underwent cardiac surgery. Adipogenesis was induced at 117, 200, or 295 mg/dl glucose, with or without macrophage‐conditioned medium (MCM). Expression of adiponectin, GLUT‐4 and the insulin receptor was analyzed by real‐time PCR. The paracrine effect of stromal cells was determined in co‐cultures with endothelial cells, by exposing them to high glucose and/or MCM, and, additionally, to leukocyte‐conditioned medium from patients with myocardial infarction. The endothelial response was determined by analyzing vascular adhesion molecule expression. Our results showed a U‐shaped dose–response curve of glucose on adiponectin in EAT, but not in SAT stromal cells. Conversely, MCM reduced the adipogenesis‐induced adiponectin expression of EAT stromal cells. The presence of EAT stromal increased the inflammatory molecules of endothelial cells. This deleterious effect was emphasized in the presence of inflammatory cell‐conditioned medium from patients with myocardial infarction. Thus, high glucose and inflammatory cells reduced adipogenesis‐induced adiponectin expression of EAT stromal cells, which induced an inflammatory paracrine process in endothelial cells. This inflammatory effect was lower in presence of mature adipocytes, producers of adiponectin. These results contribute to understanding the role of EAT dysfunction on coronary atherosclerosis progression. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  September 29, 2015   doi: 10.1002/jcp.25189   open full text
• Stress‐Induced Activation of Apoptosis Signal‐Regulating Kinase 1 Promotes Osteoarthritis.
  Qian‐Shi Zhang, Gregory J. Eaton, Carol Diallo, Theresa A. Freeman.
  Journal of Cellular Physiology. September 29, 2015
  Apoptosis signal‐regulated kinase 1 (ASK1) has been shown to affect a wide range of cellular processes including stress‐related responses, cytokine and growth factor signaling, cell cycle and cell death. Recently, we reported that lack of ASK1 slowed chondrocyte hypertrophy, terminal differentiation and apoptosis resulting in an increase in trabecular bone formation. Herein, we investigated the role of ASK1 in the pathogenesis of osteoarthritis (OA). Immunohistochemistry performed on articular cartilage samples from patients with OA showed ASK1 expression increased with OA severity. In vitro analysis of chondrocyte hypertrophy, maturation and ASK1 signaling in embryonic fibroblasts from ASK1 knockout (KO) and wild type (WT) mice was examined. Western analysis demonstrated an increase in ASK1 signaling commensurate with chondrogenic maturation during differentiation or in response to stress by the cytokines, tumor necrosis factor alpha or interleukin 1 beta in WT, but not in ASK1 KO embryonic fibroblasts. Surgically induced moderate or severe OA or OA due to natural aging in WT and ASK1 KO mice was assessed by microCT of subchondral bone, immunohistochemistry, histology, and OARSI scoring. Immunohistochemistry, microCT and OARSI scoring all indicated that the lack of ASK1 protected against OA joint degeneration, both in surgically induced OA and in aging mice. We propose that the ASK1 MAP kinase signaling cascade is an important regulator of chondrocyte terminal differentiation and inhibitors of this pathway could be useful for slowing chondrocyte maturation and cell death observed with OA progression. J. Cell. Physiol. 9999: 1–10, 2015. © 2015 Wiley Periodicals, Inc.

  September 29, 2015   doi: 10.1002/jcp.25186   open full text
• Alterations of the Plasma Peptidome Profiling in Colorectal Cancer Progression.
  Chiara Bedin, Sara Crotti, Eugenio Ragazzi, Salvatore Pucciarelli, Lisa Agatea, Ennio Tasciotti, Mauro Ferrari, Pietro Traldi, Flavio Rizzolio, Antonio Giordano, Donato Nitti, Marco Agostini.
  Journal of Cellular Physiology. September 29, 2015
  Early detection of colorectal cancer (CRC) remains a challenge. It has been highlighted that the pathological alterations within an organ and tissues might be reflected in serum or plasma proteomic/peptidic patterns. The aim of the study was to follow the changes in the plasma peptides associated to colorectal cancer progression by mass spectrometry. This study included 27 adenoma, 67 CRC (n = 33 I–II stage and n = 34 III–IV stage), 23 liver metastasis from CRC patients and 34 subjects disease‐free as controls. For plasma peptides analysis, samples purification was performed on the Nanoporous Silica Chips technology followed by matrix‐assisted laser desorption/ionisation‐time of flight analysis. Since the high complexity of the obtained dataset, multivariate statistical analysis, and discriminant pattern recognition were performed for study groups classification. Forty‐four of 88 ionic species were successfully identified as fragments of peptides and proteins physiologically circulating in the blood and belonging to immune and coagulation systems and inflammatory mediators. Many peptides clustered into sets of overlapping sequences with ladder‐like truncation clearly associated to proteolytic processes of both endo‐ and exoproteases activity. Comparing to controls, a different median ion intensity of the group‐type fragments distribution was observed. Moreover, the degradation pattern obtained by proteolytic cleavage was different into study groups. This pattern was specific and characteristic of each group: controls, colon tumour disease (including adenoma and CRC), and liver metastasis, revealing a role as biomarker in early diagnosis and prognosis. Our findings highlighted peculiar changes in protease activity characteristic of CRC progression from pre‐cancer lesion to metastatic disease. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  September 29, 2015   doi: 10.1002/jcp.25196   open full text
• Downhill Running Excessive Training Inhibits Hypertrophy in Mice Skeletal Muscles with Different Fiber Type Composition.
  Alisson L. da Rocha, Bruno C. Pereira, José R. Pauli, Claudio T. de Souza, Giovana R. Teixeira, Fábio S. Lira, Dennys E. Cintra, Eduardo R. Ropelle, Carlos R.B. Júnior, Adelino S.R. da Silva.
  Journal of Cellular Physiology. September 29, 2015
  The aim of this study was to verify the effects of running overtraining protocols performed in downhill, uphill, and without inclination on the proteins related to hypertrophy signaling pathway in extensor digitorum longus (EDL) and soleus of C57BL/6 mice. We also performed histological and stereological analyses. Rodents were divided into control (CT; sedentary mice), overtrained by downhill running (OTR/down), overtrained by uphill running (OTR/up), and overtrained by running without inclination (OTR). The incremental load, exhaustive, and grip force tests were used as performance evaluation parameters. 36 h after the grip force test, EDL and soleus were removed and immediately used for immunoblotting analysis or stored at −80°C for histological and stereological analyses. For EDL, OTR/down decreased the protein kinase B (Akt) and tuberous sclerosis protein 2 (TSC2) phosphorylation (p), and increased myostatin, receptor‐activated Smads (pSMAD2‐3), and insulin receptor substrate‐1 (pIRS‐1; Ser307/636). OTR/down also presented low and high relative proportions of cytoplasm and connective tissue, respectively. OTR/up increased the mammalian target of rapamycin (pmTOR), 70‐kDa ribosomal protein S6 kinase 1 (pS6K1) and pSMAD2‐3, and decreased pTSC2. OTR decreased pTSC2 and increased pIRS‐1 (Ser636). For soleus, OTR/down increased S6 ribosomal protein (pS6RP) and pSMAD2‐3, and decreased pIRS‐1 (Ser639). OTR/up decreased pS6K1, pS6RP and pIRS‐1 (Ser639), and increased pTSC2 (Ser939), and pSMAD2‐3. OTR increased pS6RP, 4E‐binding protein‐1 (p4E‐BP1), pTSC2 (Ser939), and pSMAD2‐3, and decreased pIRS‐1 (Ser639). In summary, OTR/down inhibited the skeletal muscle hypertrophy with concomitant signs of atrophy in EDL. The effects of OTR/up and OTR depended on the analyzed skeletal muscle type. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  September 29, 2015   doi: 10.1002/jcp.25197   open full text
• Mesenchymal Stromal Cells Differentiating to Adipocytes Accumulate Autophagic Vesicles Instead of Functional Lipid Droplets.
  Alexandra T. Gruia, Maria Suciu, Lucian Barbu‐Tudoran, Seyed Mohammad Reza Azghadi, Mirabela I. Cristea, Dragos V. Nica, Adrian Vaduva, Danina Muntean, Ani Aurora Mic, Felix A. Mic.
  Journal of Cellular Physiology. September 22, 2015
  Adult bone marrow mesenchymal stromal cells (BMSCs) can easily be differentiated into a variety of cells. In vivo transplantation of BMSCs‐differentiated cells has had limited success, suggesting that these cells may not be fully compatible with the cells they are intended to replace in vivo. We investigated the structural and functional features of BMSCs‐derived adipocytes as compared with adipocytes from adipose tissue, and the structure and functionality of lipid vesicles formed during BMSCs differentiation to adipocytes. Gas chromatography–mass spectrometry showed fatty acid composition of BMSCs‐derived adipocytes and adipocytes from the adipose tissue to be very different, as is the lipid rafts composition, caveolin‐1 expression, caveolae distribution in their membranes, and the pattern of expression of fatty acid elongases. Confocal microscopy confirmed the absence from BMSCs‐derived adipocytes of markers of lipid droplets. BMSCs‐derived adipocytes cannot convert deuterated glucose into deuterated species of fatty acids and cannot uptake the deuterated fatty acid–bovine serum albumin complexes from the culture medium, suggesting that intra‐cellular accumulation of lipids does not occur by lipogenesis. We noted that BMSCs differentiation to adipocytes is accompanied by an increase in autophagy. Autophagic vesicles accumulate in the cytoplasm of BMSCs‐derived adipocytes and their size and distribution resembles that of Nile Red‐stained lipid vesicles. Stimulation of autophagy in BMSCs triggers the intra‐cellular accumulation of lipids, while inhibition of autophagy prevents this accumulation. In conclusion, differentiation of BMSCs‐derived adipocytes leads to intra‐cellular accumulation of autophagic vesicles rather than functional lipid droplets, suggesting that these cells are not authentic adipocytes. J. Cell. Physiol. 9999: 1–13, 2015. © 2015 Wiley Periodicals, Inc.

  September 22, 2015   doi: 10.1002/jcp.25177   open full text
• Tumor Microenvironment Versus Cancer Stem Cells in Cholangiocarcinoma: Synergistic Effects?
  Maurizio Romano, Francesco De Francesco, Enrico Gringeri, Antonio Giordano, Giuseppe A. Ferraro, Marina Di Domenico, Umberto Cillo.
  Journal of Cellular Physiology. September 22, 2015
  Cholangiocarcinoma (CCAs) may be defined as tumors that derived from the biliary tree with the differentiation in the biliary epithelial cells. This tumor is malignant, extremely aggressive with a poor prognosis. It can be treated surgically and its pathogenesis is poorly understood. The tumor microenvironment (TME) is a very important factor in the regulation of tumor angiogenesis, invasion, and metastasis. Besides cancer stem cells (CSCs) can modulate tumor growth, stroma formation, and migratory capability. The initial stage of tumorigenesis is characterized by genetic mutations and epigenetic alterations due to intrinsic factors which lead to the generation of oncogenes thus inducing tumorigenesis. CSCs may result from precancerous stem cells, cell de‐differentiation, normal stem cells, or an epithelial‐mesenchymal transition (EMT). CSCs have been found in the cancer niche, and EMT may occur early within the tumor microenvironment. Previous studies have demonstrated evidence of cholangiocarcinoma stem cells (CD133, CD24, EpCAM, CD44, and others) and the presence of these markers has been associated with malignant potential. The interaction between TME and cholangiocarcinoma stem cells via signaling mediators may create an environment that accommodates tumor growth, yielding resistance to cytotoxic insults (chemotherarapeutic). While progress has been made in the understanding of the mechanisms, the interactions in the tumorigenic process still remain a major challenge. Our review, addresses recent concepts of TME‐CSCs interaction and will emphasize the importance of early detection with the use of novel diagnostic mechanisms such as CCA–CSC biomarkers and the importance of tumor stroma to define new treatments. J. Cell. Physiol. 9999: 1–9, 2015. © 2015 Wiley Periodicals, Inc.

  September 22, 2015   doi: 10.1002/jcp.25190   open full text
• HhAntag, a Hedgehog Signaling Antagonist, Suppresses Chondrogenesis and Modulates Canonical and Non‐Canonical BMP Signaling.
  Christina Mundy, Adebayo Bello, Federica Sgariglia, Eiki Koyama, Maurizio Pacifici.
  Journal of Cellular Physiology. September 22, 2015
  Chondrogenesis subtends the development of most skeletal elements and involves mesenchymal cell condensations differentiating into growth plate chondrocytes that proliferate, undergo hypertrophy, and are replaced by bone. In the pediatric disorder Hereditary Multiple Exostoses, however, chondrogenesis occurs also at ectopic sites and causes formation of benign cartilaginous tumors—exostoses—near the growth plates. No treatment is currently available to prevent or reverse exostosis formation. Here, we asked whether chondrogenesis could be stopped by targeting the hedgehog pathway, one of its major regulators. Micromass cultures of limb mesenchymal cells were treated with increasing amounts of the hedgehog inhibitor HhAntag or vehicle. The drug effectively blocked chondrogenesis and did so in a dose‐dependent manner as monitored by: alcian blue‐positive cartilage nodule formation; gene expression of cartilage marker genes; and reporter activity in Gli1‐LacZ cell cultures. HhAntag blocked chondrogenesis even when the cultures were co‐treated with bone morphogenetic protein 2 (rhBMP‐2), a strong pro‐chondrogenic factor. Immunoblots showed that HhAntag action included modulation of canonical (pSmad1/5/8) and non‐canonical (pp38) BMP signaling. In cultures co‐treated with HhAntag plus rhBMP‐2, there was a surprising strong up‐regulation of pp38 levels. Implantation of rhBMP‐2‐coated beads near metacarpal elements in cultured forelimb explants induced formation of ectopic cartilage that however, was counteracted by HhAntag co‐treatment. Collectively, our data indicate that HhAntag inhibits not only hedgehog signaling, but also modulates canonical and non‐canonical BMP signaling and blocks basal and rhBMP2‐stimulated chondrogenesis, thus representing a potentially powerful drug‐based strategy to counter ectopic cartilage growth or induce its involution. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  September 22, 2015   doi: 10.1002/jcp.25192   open full text
• TRPV4 Channels in Human White Adipocytes: Electrophysiological Characterization and Regulation by Insulin.
  Julio C. Sánchez, Ricardo A. Rivera, Laura V. Muñoz.
  Journal of Cellular Physiology. September 18, 2015
  Intracellular calcium homeostasis in adipocytes is important for the regulation of several functions and is involved in pathological changes in obesity and other associated diseases. Transient Receptor Potential Vanilloid 4 (TRPV4) channels are an important route for calcium entry that operates in a variety of cells and intervenes in a number of functions. In this study, the expression and operation of TRPV4 channels in human cultured adipocytes was evaluated using RT‐PCR, Western blotting, the whole‐cell patch‐clamp technique and fluorescence measurements to characterize these channels and determine intracellular calcium responses. Both the hypoosmolarity and 4alpha‐phorbol‐didecanoate (4αPDD), a specific TRPV4 agonist, induced a similar HC‐067047‐sensitive current, which was predominantly inward, and an intracellular Ca2+ concentration increase, which was exclusively dependent on extracellular calcium, and membrane depolarization. The current had a reverse potential of +31 ± 6 mV and exhibited preferential permeability to Ca2+. Insulin, which regulates metabolic homeostasis in adipocytes, attenuated the TRPV4‐mediated effects. These results confirm the function of TRPV4 in human cultured adipocytes and its regulation by insulin. J. Cell. Physiol. 9999: 1–10, 2015. © 2015 Wiley Periodicals, Inc.

  September 18, 2015   doi: 10.1002/jcp.25187   open full text
• Mitotic Inheritance of mRNA Facilitates Translational Activation of the Osteogenic‐Lineage Commitment Factor Runx2 in Progeny of Osteoblastic Cells.
  Nelson Varela, Alejandra Aranguiz, Carlos Lizama, Hugo Sepulveda, Marcelo Antonelli, Roman Thaler, Ricardo D. Moreno, Martin Montecino, Gary S. Stein, Andre J. van Wijnen, Mario Galindo.
  Journal of Cellular Physiology. September 18, 2015
  Epigenetic mechanisms mediate the acquisition of specialized cellular phenotypes during tissue development, maintenance and repair. When phenotype‐committed cells transit through mitosis, chromosomal condensation counteracts epigenetic activation of gene expression. Subsequent post‐mitotic re‐activation of transcription depends on epigenetic DNA and histone modifications, as well as other architecturally bound proteins that “bookmark” the genome. Osteogenic lineage commitment, differentiation and progenitor proliferation require the bone‐related runt‐related transcription factor Runx2. Here, we characterized a non‐genomic mRNA mediated mechanism by which osteoblast precursors retain their phenotype during self‐renewal. We show that osteoblasts produce maximal levels of Runx2 mRNA, but not protein, prior to mitotic cell division. Runx2 mRNA partitions symmetrically between daughter cells in a non‐chromosomal tubulin‐containing compartment. Subsequently, transcription‐independent de novo synthesis of Runx2 protein in early G1 phase results in increased functional interactions of Runx2 with a representative osteoblast‐specific target gene (osteocalcin/BGLAP2) in chromatin. Somatic transmission of Runx2 mRNAs in osteoblasts and osteosarcoma cells represents a versatile mechanism for translational rather than transcriptional induction of this principal gene regulator to maintain osteoblast phenotype identity after mitosis. J. Cell. Physiol. 9999: 1–14, 2015. © 2015 Wiley Periodicals, Inc.

  September 18, 2015   doi: 10.1002/jcp.25188   open full text
• Mechanical Loading Stimulates Expression of Collagen Cross‐Linking Associated Enzymes in Periodontal Ligament.
  Masaru Kaku, Juan Marcelo Rosales Rocabado, Megumi Kitami, Takako Ida, Yosuke Akiba, Mitsuo Yamauchi, Katsumi Uoshima.
  Journal of Cellular Physiology. September 18, 2015
  Type I collagen, a major extracellular component of the periodontal ligament (PDL), is post‐translationally modified by a series of specific enzymes. Among the collagen‐modifying enzymes, lysyl oxidase (LOX) is essential to initiate collagen cross‐linking and lysyl hydroxylases (LHs) to regulate the cross‐linking pathways that are important for tissue specific mechanical properties. The purpose of this study was to investigate the effects of mechanical loading on the expression of collagen‐modifying enzymes and subsequent tissue changes in PDL. Primary human PDL cells were subjected to mechanical loading in a 3D collagen gel, and gene expression and collagen component were analyzed. Wistar rats were subjected to excessive occlusal loading with or without intra‐peritoneal injection of a LOX inhibitor, β‐aminopropionitrile (BAPN). Upon mechanical loading, gene expression of LH2 and LOX was significantly elevated, while that of COL1A2 was not affected on hPDL‐derived cells. The mechanical loading also elevated formation of collagen α‐chain dimers in 3D culture. The numbers of LH2 and LOX positive cells in PDL were significantly increased in an excessive occlusal loading model. Notably, an increase of LH2‐positive cells was observed only at the bone‐side of PDL. Intensity of picrosirius red staining was increased by excessive occlusal loading, but significantly diminished by BAPN treatment. These results demonstrated that mechanical loading induced collagen maturation in PDL by up‐regulating collagen‐modifying enzymes and subsequent collagen cross‐linking which are important for PDL tissue maintenance. J. Cell. Physiol. 9999: 1–8, 2015. © 2015 Wiley Periodicals, Inc.

  September 18, 2015   doi: 10.1002/jcp.25184   open full text
• Alpha‐Catulin Co‐Localizes With Vimentin Intermediate Filaments and Functions in Pulmonary Vascular Endothelial Cell Migration via ROCK.
  Michael D. Bear, Tiegang Liu, Shereen Abualkhair, Maher A. Ghamloush, Nicholas S. Hill, Ioana Preston, Barry L. Fanburg, Usamah S. Kayyali, Deniz Toksoz.
  Journal of Cellular Physiology. September 17, 2015
  The ubiquitous α‐catulin acts as a scaffold for distinct signalosomes including RhoA/ROCK; however, its function is not well understood. While α‐catulin has homology to the cytoskeletal linkers α‐catenin and vinculin, it appears to be functionally divergent. Here we further investigated α‐catulin function in pulmonary vascular endothelial cells (VEC) on the premise that α‐catulin has a unique cytoskeletal role. Examination of endogenous α‐catulin intracellular localization by immunofluorescence revealed a highly organized cytosolic filamentous network suggestive of a cytoskeletal system in a variety of cultured VEC. Double‐immunofluorescence analyses of VEC showed endogenous α‐catulin co‐localization with vimentin intermediate filaments. Similar to vimentin, α‐catulin was found to distribute into detergent‐soluble and ‐insoluble fractions. Treatment of VEC with withaferinA, an agent that targets vimentin filaments, disrupted the α‐catulin network distribution and altered α‐catulin solubility. Vimentin participates in cell migration, and withaferinA was found to inhibit VEC migration in vitro; similarly, α‐catulin knock‐down reduced VEC migration. Based on previous reports showing that ROCK modulates vimentin, we found that ROCK depletion attenuated VEC migration; furthermore, α‐catulin depletion was shown to reduce ROCK‐induced signaling. These findings indicate that α‐catulin has a unique function in co‐localization with vimentin filaments that contributes to VEC migration via a pathway that may involve ROCK signaling. J. Cell. Physiol. 9999: 1–10, 2015. © 2015 Wiley Periodicals, Inc.

  September 17, 2015   doi: 10.1002/jcp.25185   open full text
• Neurotransmitters and Neuropeptides: New Players in the Control of Islet of Langerhans' Cell Mass and Function.
  Eliana S. Di Cairano, Stefania Moretti, Paola Marciani, Vellea Franca Sacchi, Michela Castagna, Alberto Davalli, Franco Folli, Carla Perego.
  Journal of Cellular Physiology. September 10, 2015
  Islets of Langerhans control whole body glucose homeostasis, as they respond, releasing hormones, to changes in nutrient concentrations in the blood stream. The regulation of hormone secretion has been the focus of attention for a long time because it is related to many metabolic disorders, including diabetes mellitus. Endocrine cells of the islet use a sophisticate system of endocrine, paracrine and autocrine signals to synchronize their activities. These signals provide a fast and accurate control not only for hormone release but also for cell differentiation and survival, key aspects in islet physiology and pathology. Among the different categories of paracrine/autocrine signals, this review highlights the role of neurotransmitters and neuropeptides. In a manner similar to neurons, endocrine cells synthesize, accumulate, release neurotransmitters in the islet milieu, and possess receptors able to decode these signals. In this review, we provide a comprehensive description of neurotransmitter/neuropetide signaling pathways present within the islet. Then, we focus on evidence supporting the concept that neurotransmitters/neuropeptides and their receptors are interesting new targets to preserve β‐cell function and mass. A greater understanding of how this network of signals works in physiological and pathological conditions would advance our knowledge of islet biology and physiology and uncover potentially new areas of pharmacological intervention. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  September 10, 2015   doi: 10.1002/jcp.25176   open full text
• Beta Adrenergic Receptor Stimulation Suppresses Cell Migration in Association with Cell Cycle Transition in Osteoblasts—Live Imaging Analyses Based on FUCCI System.
  Sakie Katsumura, Yoichi Ezura, Yayoi Izu, Jumpei Shirakawa, Atsushi Miyawaki, Kiyoshi Harada, Masaki Noda.
  Journal of Cellular Physiology. September 09, 2015
  Osteoporosis affects over 20 million patients in the United States. Among those, disuse osteoporosis is serious as it is induced by bed‐ridden conditions in patients suffering from aging‐associated diseases including cardiovascular, neurological, and malignant neoplastic diseases. Although the phenomenon that loss of mechanical stress such as bed‐ridden condition reduces bone mass is clear, molecular bases for the disuse osteoporosis are still incompletely understood. In disuse osteoporosis model, bone loss is interfered by inhibitors of sympathetic tone and adrenergic receptors that suppress bone formation. However, how beta adrenergic stimulation affects osteoblastic migration and associated proliferation is not known. Here we introduced a live imaging system, fluorescent ubiquitination‐based cell cycle indicator (FUCCI), in osteoblast biology and examined isoproterenol regulation of cell cycle transition and cell migration in osteoblasts. Isoproterenol treatment suppresses the levels of first entry peak of quiescent osteoblastic cells into cell cycle phase by shifting from G1/G0 to S/G2/M and also suppresses the levels of second major peak population that enters into S/G2/M. The isoproterenol regulation of osteoblastic cell cycle transition is associated with isoproterenol suppression on the velocity of migration. This isoproterenol regulation of migration velocity is cell cycle phase specific as it suppresses migration velocity of osteoblasts in G1 phase but not in G1/S nor in G2/M phase. Finally, these observations on isoproterenol regulation of osteoblastic migration and cell cycle transition are opposite to the PTH actions in osteoblasts. In summary, we discovered that sympathetic tone regulates osteoblastic migration in association with cell cycle transition by using FUCCI system. J. Cell. Physiol. 231: 496–504, 2016. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25096   open full text
• Circulating MicroRNA and Long Noncoding RNA as Biomarkers of Cardiovascular Diseases.
  Qiang Shi, Xi Yang.
  Journal of Cellular Physiology. September 09, 2015
  Although >85% of the human genome is transcribed, only <2% is transcribed into protein‐coding RNA (messenger RNA, mRNA). Many thousands of noncoding RNAs are transcribed and recognized as functional RNAs with diverse sizes, structures, and biological functions. Based on size, noncoding RNA can be generally divided into two subgroups: short noncoding RNA (<200 nucleotides including microRNA or miRNA) and long noncoding RNA (lncRNA, >200 nucleotides). It is now clear that these RNAs fulfil critical roles as transcriptional and post‐transcriptional regulators and as guides of chromatin‐modifying complexes. Although not translated into protein, noncoding RNAs can regulate cardiac function through diverse mechanisms and their dysregulation is increasingly linked with cardiovascular pathophysiology. Furthermore, a series of recent studies have discovered that noncoding RNAs can be found in the bloodstream and some species are remarkably stable. This has raised the possibility that such noncoding RNAs may be measured in body fluids and serve as novel diagnostic biomarkers. Here, we summarize the current knowledge of noncoding RNAs’ function and biomarker potential in cardiac diseases, concentrating mainly on circulating miRNAs and lncRNAs. J. Cell. Physiol. 9999: 1–5, 2015. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25174   open full text
• Downregulation of Long Noncoding RNA Meg3 Affects Insulin Synthesis and Secretion in Mouse Pancreatic Beta Cells.
  LiangHui You, Ning Wang, DanDan Yin, LinTao Wang, FeiYan Jin, YaNan Zhu, QingXin Yuan, Wei De.
  Journal of Cellular Physiology. September 09, 2015
  Increasing evidence indicates that long noncoding RNAs (lncRNAs) are involved in diverse biological process. Mouse maternal expressed gene 3 (Meg3) is an imprinted gene and essential for development. Here, we explored the relationship between Meg3 and the function of mouse beta cells in vitro and in vivo. Real‐time PCR analyses revealed that Meg3 was more abundantly expressed in Balb/c mouse islets than exocrine glands. Moreover, the expression of Meg3 in islets was decreased in T1DM (NOD female mice) and T2DM (db/db mice) models. Meg3 expression was modulated dynamically by glucose in Min6 cells and isolated mouse islets. The function role of Meg3 was investigated in Min6 cells and normal mouse by knockdown of Meg3 using small interfering RNA. After suppression of Meg3 expression in vitro, insulin synthesis and secretion were impaired and the rate of beta cells apoptosis was increased. Moreover, knockdown of Meg3 in vivo led to the impaired glucose tolerance and decreased insulin secretion, consisted with the reduction of insulin positive cells areas by immunochemistry assays. Notably, islets from Meg3 interference groups showed significant decrease of Pdx‐1 and MafA expression in mRNA and protein levels. These results indicate that Meg3 may function as a new regulator of maintaining beta cells identity via affecting insulin production and cell apoptosis. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25175   open full text
• Modulating Intrafollicular Hormonal Milieu in Controlled Ovarian Stimulation: Insights From PPAR Expression in Human Granulosa Cells.
  Carla Tatone, Elisabetta Benedetti, Maurizio Vitti, Giovanna Di Emidio, Rosanna Ciriminna, Maria Elena Vento, Vito Cela, Placido Borzì, Gaspare Carta, Monica Lispi, Anna Maria Cimini, Paolo Giovanni Artini,.
  Journal of Cellular Physiology. September 09, 2015
  Controlled ovarian stimulation (COS) leading to ovulation of multiple follicles is a crucial aspect of biomedical infertility care. Nevertheless, biomarkers useful for COS management are still lacking. Peroxisome proliferator‐activated receptors (PPARs) are nuclear hormone receptors relevant to steroid metabolism in granulosa cells (GCs). We investigated whether PPARs and their steroidogenic targets were differentially expressed in GCs differentiated under different recombinant or urinary gonadotropin preparations. GCs from women subjected to COS with r‐hFSH, r‐hFSH/r‐hLH, or hMG‐HP were processed to assess expression of PPARα, PPARβ/δ, PPARγ, and steroidogenic enzymes under PPAR modulation. As an evidence of their activation, all PPAR isotypes with their coactivators, the retinoic‐X‐receptors (RXRs), localized in the nucleus. When GCs from r‐hFSH/r‐hLH group were compared with r‐hFSH, a significant reduction of PPARα protein was observed. By contrast, an increase of PPARβ/δ at both protein and mRNA levels along with that of PPARγ protein were detected. The steroidogenic enzymes 17βHSD IV, 3βHSD II, and HMG‐CoA red were downregulated in the r‐hFSH/r‐hLH group in comparison to r‐hFSH unlike CYP19A1 that remained unchanged. In GCs from urinary FSH‐LH stimulation (hMG‐HP), PPARα was more expressed in comparison with r‐hFSH/r‐hLH group. Likewise, 3βHSD II and 17βHSD IV were increased suggesting that hMG‐HP partially mimicked r‐hFSH/r‐hLH effects. In summary, transcript analysis associated to protein investigation revealed differential effects of COS protocols on PPARs and their steroidogenic targets in relation to LH and gonadotropin source. These observations candidate PPARs as new biomarkers of follicle competence opening new hypotheses on COS effects on ovarian physiology. J. Cell. Physiol. 9999: 1–7, 2015. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25182   open full text
• Kidney Injury Molecule‐1 Enhances Endocytosis of Albumin in Renal Proximal Tubular Cells.
  Xueying Zhao, Chen Jiang, Rebecca Olufade, Dong Liu, Nerimiah Emmett.
  Journal of Cellular Physiology. September 09, 2015
  Receptor‐mediated endocytosis plays an important role in albumin reabsorption by renal proximal tubule epithelial cells. Kidney injury molecule‐1 (KIM‐1) is a scavenger receptor that is upregulated on the apical membrane of proximal tubules in proteinuric kidney disease. In this study, we examined the cellular localization and functional role of KIM‐1 in cultured renal tubule epithelial cells (TECs). Confocal immunofluorescence microscopy reveals intracellular and cell surface localization of KIM‐1 in primary renal TECs. Albumin stimulation resulted in a redistribution of KIM‐1 and tight junction protein zonula occludens‐1 in primary TEC monolayer. An increase in albumin internalization was observed in both primary TECs expressing endogenous KIM‐1 and rat kidney cell line (NRK‐52E) overexpressing exogenous KIM‐1. KIM‐1‐induced albumin accumulation was abolished by its specific antibody. Moreover, endocytosed KIM‐1 and its cargo proteins were delivered from endosomes to lysosomes for degradation in a clathrin‐dependent pathway. Supportive evidence includes (1) detection of KIM‐1 in Rab5‐positive early endosomes, Rab7‐positive late endosomes/multivesicular bodies, and LAMP1‐positive lysosomes, (2) colocalization of KIM‐1 and clathrin in the intracellular vesicles, and (3) blockade of KIM‐1‐mediated albumin internalization by chlorpromazine, an inhibitor of clathrin‐dependent endocytosis. KIM‐1 expression was upregulated by albumin but downregulated by transforming growth factor‐β1. Taken together, our data indicate that KIM‐1 increases albumin endocytosis in renal tubule epithelial cells, at least partially via a clathrin‐dependent mechanism. J. Cell. Physiol. 9999: 1–12, 2015. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25181   open full text
• Osteosarcoma Stem Cells Have Active Wnt/β‐catenin and Overexpress SOX2 and KLF4.
  Sara R. Martins‐Neves, Willem E. Corver, Daniela I. Paiva‐Oliveira, Brendy E. W. M. van den Akker, Inge H. Briaire‐de‐Bruijn, Judith V. M. G. Bovée, Célia M. F. Gomes, Anne‐Marie Cleton‐Jansen.
  Journal of Cellular Physiology. September 09, 2015
  Osteosarcoma is a bone tumor, displaying significant cellular and histological heterogeneity and a complex genetic phenotype. Although multiple studies strongly suggest the presence of cancer stem cells in osteosarcoma, a consensus on their characterization is still missing. We used a combination of functional assays (sphere‐forming, Aldefluor, and side‐population) for identification of cancer stem cell populations in osteosarcoma cell lines. Expression of stemness‐related transcription factors, quiescent nature, in vivo tumorigenicity, and Wnt/β‐catenin activation were evaluated. We show that different cancer stem cell populations may co‐exist in osteosarcoma cell lines exhibiting distinct functional properties. Osteosarcoma spheres are slowly‐proliferating populations, overexpress SOX2, and KLF4 stemness‐related genes and have enhanced tumorigenic potential. Additionally, spheres show specific activation of Wnt/β‐catenin signaling as evidenced by increased nuclear β‐catenin, TCF/LEF activity, and AXIN2 expression, in a subset of the cell lines. Aldefluor‐positive populations were detected in all osteosarcoma cell lines and overexpress SOX2, but not KLF4. The side‐population phenotype is correlated with ABCG2 drug‐efflux transporter expression. Distinct functional methods seem to identify cancer stem cells with dissimilar characteristics. Intrinsic heterogeneity may exist within osteosarcoma cancer stem cells and can have implications on the design of targeted therapies aiming to eradicate these cells within tumors. J. Cell. Physiol. 9999: 1–11, 2015. © 2015 Wiley Periodicals, Inc.

  September 09, 2015   doi: 10.1002/jcp.25179   open full text
• Epigenetic Priming Confers Direct Cell Trans‐Differentiation from Adipocyte to Osteoblast in a Transgene‐Free State.
  Young‐Dan Cho, Han‐Sol Bae, Dong‐Seol Lee, Won‐Joon Yoon, Kyung‐Mi Woo, Jeong‐Hwa Baek, Gene Lee, Joo‐Cheol Park, Young Ku, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. September 03, 2015
  The bone marrow of healthy individuals is primarily composed of osteoblasts and hematopoietic cells, while that of osteoporosis patients has a larger portion of adipocytes. There is evidence that the epigenetic landscape can strongly influence cell differentiation. We have shown that it is possible to direct the trans‐differentiation of adipocytes to osteoblasts by modifying the epigenetic landscape with a DNA methyltransferase inhibitor (DNMTi), 5′‐aza‐dC, followed by Wnt3a treatment to signal osteogenesis. Treating 3T3‐L1 adipocytes with 5′‐aza‐dC induced demethylation in the hypermethylated CpG regions of bone marker genes; subsequent Wnt3a treatment drove the cells to osteogenic differentiation. When old mice with predominantly adipose marrow were treated with both 5′‐aza‐dC and Wnt3a, decreased fatty tissue and increased bone volume were observed. Together, our results indicate that epigenetic modification permits direct programming of adipocytes into osteoblasts in a mouse model of osteoporosis, suggesting that this approach could be useful in bone tissue‐engineering applications. This article is protected by copyright. All rights reserved

  September 03, 2015   doi: 10.1002/jcp.25183   open full text
• Selective β2‐AR Blockage Suppresses Colorectal Cancer Growth Through Regulation of EGFR‐Akt/ERK1/2 Signaling, G1‐Phase Arrest, and Apoptosis.
  Chih‐Chien Chin, Jhy‐Ming Li, Kam‐Fai Lee, Yun‐Ching Huang, Kuan‐Chieh Wang, Hsiao‐Ching Lai, Chih‐Chung Cheng, Yi‐Hung Kuo, Chung‐Sheng Shi.
  Journal of Cellular Physiology. September 01, 2015
  The stress‐upregulated catecholamines‐activated β1‐ and β2‐adrenergic receptors (β1/2‐ARs) have been shown to accelerate the progression of cancers such as colorectal cancer (CRC). We investigated the underlying mechanism of the inhibition of β1/2‐ARs signaling for the treatment of CRC and elucidated the significance of β2‐AR expression in CRC in vitro and in clinical samples. The impacts of β1/2‐AR antagonists in CRC in vitro and CRC‐xenograft in vivo were examined. We found that repression of β2‐AR but not β1‐AR signaling selectively suppressed cell viability, induced G1‐phase cell cycle arrest, caused both intrinsic and extrinsic pathways‐mediated apoptosis of specific CRC cells and inhibited CRC‐xenograft growth in vivo. Moreover, the expression of β2‐AR was not consistent with the progression of CRC in vitro or in clinical samples. Our data evidence that the expression profiles, signaling, and blockage of β2‐AR have a unique pattern in CRC comparing to other cancers. β2‐AR antagonism selectively suppresses the growth of CRC accompanying active β2‐AR signaling, which potentially carries wild‐type KRAS, in vitro and in vivo via the inhibition of β2‐AR transactivated EFGR‐Akt/ERK1/2 signaling pathway. Thus, β2‐AR blockage might be a potential therapeutic strategy for combating the progressions of β2‐AR‐dependent CRC. J. Cell. Physiol. 231: 459–472, 2016. © 2015 Wiley Periodicals, Inc.

  September 01, 2015   doi: 10.1002/jcp.25092   open full text
• Comparison of Dasatinib, Nilotinib, and Imatinib in the Treatment of Chronic Myeloid Leukemia.
  Roberto Ciarcia, Sara Damiano, Maria Valeria Puzio, Serena Montagnaro, Francesco Pagnini, Carmen Pacilio, Giuseppe Caparrotti, Cristiana Bellan, Tiziana Garofano, Maria Sole Polito, Antonio Giordano, Salvatore Florio.
  Journal of Cellular Physiology. September 01, 2015
  To overcome the drug resistance phenomenon induced by Imatibib (IM), in clinical practice, are often used second generation of tyrosine kinase inhibitors as Nilotinib (NIL); a such potent inhibitor of the BCR/ABL kinase and Dasatinib (DAS), a inhibitor of BCR/ABL kinase, and inhibitor SrC family kinase. In this study we evaluated the in vivo effect of DAS, NIL, and IM on intracellular calcium concentration, oxidative stress, and apoptosis in peripheral blood leukocytes of 45 newly diagnosed patients with chronic myeloid leukaemia (CML‐PBM). Our data demonstrated that treatment with DAS and NIL showed an higher modulating potential than IM on intracellular calcium concentration by inhibiting the thapsigargin, a sarcoplasmic/endoplasmic reticulum Ca2+‐ATPase (SERCA) inhibitor, and Lithium (Li) an inositol 1,4,5‐triphosphate (InsP3) receptor inhibitor activities. Moreover our data demonstrated that NIL and DAS have significantly increased apoptosis more than IM by involving both intracellular calcium signaling as well as oxidative stress. The acquisition of the oxidative stress and calcium channels receptors values data could help the hematologist to modulate and improve the treatment of chronic myeloid leukaemia (CML) pathology. J. Cell. Physiol. 9999: 1–8, 2015. © 2015 Wiley Periodicals, Inc.

  September 01, 2015   doi: 10.1002/jcp.25118   open full text
• Vitamin D Induces Cyclooxygenase 2 Dependent Prostaglandin E2 Synthesis in HaCaT Keratinocytes.
  Amiram Ravid, Ohad Shenker, Efrat Buchner‐Maman, Carmela Rotem, Ruth Koren.
  Journal of Cellular Physiology. September 01, 2015
  The active metabolite of vitamin D calcitriol and its analogs are well‐known for their anti‐inflammatory action in the skin, while their main side effect associated with topical treatment of inflammatory disorders is irritant contact dermatitis. Prostaglandin E2 (PGE2) is pro‐inflammatory at the onset of inflammation and anti‐inflammatory at its resolution. We hypothesized that induction of PGE2 synthesis by calcitriol in epidermal keratinocytes may contribute both to its pro‐inflammatory and anti‐inflammatory effects on the skin. Treatment of human immortalized HaCaT keratinocytes with calcitriol (3–100 nM, 2–24 h) increased PGE2 production due to increased mRNA and protein expression of COX‐2, but not to increase of COX‐1 or release of arachidonic acid. The effect of calcitriol on COX‐2 mRNA was observed also in primary human keratinocytes. The increase in COX‐2 mRNA is associated with COX‐2 transcript stabilization. Calcitriol exerts this effect by a rapid (2 h) and protein synthesis independent mode of action that is dependent on PKC and Src kinase activities. Treatment with a COX‐2 inhibitor partially prevented the attenuation of the keratinocyte inflammatory response by calcitriol. We conclude that upregulation of COX‐2 expression with the consequent increase in PGE2 synthesis may be one of the mechanisms explaining the Janus face of calcitriol as both a promoter and attenuator of cutaneous inflammation. J. Cell. Physiol. 9999: 1–7, 2015. © 2015 Wiley Periodicals, Inc.

  September 01, 2015   doi: 10.1002/jcp.25132   open full text
• G Protein‐Coupled Receptor 120 Signaling Negatively Regulates Osteoclast Differentiation, Survival, and Function.
  Hyun‐Ju Kim, Hye‐Jin Yoon, Bo Kyung Kim, Woo Youl Kang, Sook Jin Seong, Mi‐Sun Lim, Shin‐Yoon Kim, Young‐Ran Yoon.
  Journal of Cellular Physiology. September 01, 2015
  G protein‐coupled receptor 120 (GPR120) plays an important role in the regulation of inflammation and lipid metabolism. In this study, we investigated the role of GPR120 in osteoclast development and found that GPR120 regulates osteoclast differentiation, survival and function. We observed that GPR120 was highly expressed in osteoclasts compared to their precursors, bone marrow‐derived macrophages (BMMs). Activation of GPR120 by its ligand GW9508 suppressed receptor activator of NF‐ κB ligand (RANKL)‐induced osteoclast differentiation and the expression of nuclear factor of activated T cells c1 (NFATc1), a key modulator of osteoclastogenesis. GPR120 activation further inhibited the RANKL‐stimulated phosphorylation of IκBα and JNK. In addition to osteoclast differentiation, GPR120 activation increased the apoptosis of mature osteoclasts by inducing caspase‐3 and Bim expression. Activation of GPR120 also interfered with cell spreading and actin cytoskeletal organization mediated by M‐CSF but not by RANKL. Coincident with the impaired cytoskeletal organization, GPR120 activation blocked osteoclast bone resorbing activity. Furthermore, knockdown of GPR120 using small hairpin RNA abrogated all these inhibitory effects on osteoclast differentiation, survival, and function. Together, our findings identify GPR120 as a negative modulator of osteoclast development that may be an attractive therapeutic target for bone‐destructive diseases. J. Cell. Physiol. 9999: 1–8, 2015. © 2015 Wiley Periodicals, Inc.

  September 01, 2015   doi: 10.1002/jcp.25133   open full text
• Toll‐Like Receptors and Tissue Remodeling: The Pro/Cons Recent Findings.
  Alessandra Micera, Bijorn Omar Balzamino, Antonio Di Zazzo, Filippo Biamonte, Gigliola Sica, Stefano Bonini.
  Journal of Cellular Physiology. September 01, 2015
  The Toll‐like Receptor (TLR) family ensures prompt response towards pathogens, protecting the host against infections, and guarantees a realistic balance between protective and detrimental activities. Multiple regulating mechanisms characterize TLR activity that is not limited to innate and adaptive antimicrobial immune responses, as observed in the inflammatory (either infective, allergic, or autoimmune) responses associated with tissue remodeling. Following the insult and the arise of inflammatory response, tissue remodeling takes place and might develop in fibrosis, depending on microenvironment as a result of imbalanced fibroblasts (FBs) and myofibroblasts (myoFBs) activation/survival. The process is driven by an epithelial‐fibroblast‐immune cell cross‐talk. While the main FB function is the matrix metabolism for tissue homeostasis or repair, the myoFB differentiation represents a crucial step in attempting repair of injury. FBs/myoFBs provide more than structural support at site of injury, synthesizing and/or reacting to different cytokines, growth factors, neuromediators and soluble/lipid mediators. TLR‐bearing FBs/myoFBs might contribute at the innate immune level, providing a second line of protection/defense as well as being a target/effector cell of tissue remodeling. TLRs might also interfere with acute inflammation as well as with established fibrosis, triggering structural/functional changes in agreement with the genetic background, the site of lesion, the entity of associated infection, the poor blood circulation or the pharmacological treatments, all together strictly influencing tissue repair/remodeling process. This review will focus on the recent findings on TLRs at launch and long‐lasting tissue remodeling process, that strongly suggest TLRs as optional targets for future therapies. J. Cell. Physiol. 9999: 1–14, 2015. © 2015 Wiley Periodicals, Inc.

  September 01, 2015   doi: 10.1002/jcp.25124   open full text
• Cell Surface Human Airway Trypsin‐Like Protease Is Lost During Squamous Cell Carcinogenesis.
  Michael J. Duhaime, Khaliph O. Page, Fausto A. Varela, Andrew S. Murray, Michael E. Silverman, Gina L. Zoratti, Karin List.
  Journal of Cellular Physiology. August 22, 2015
  Cancer progression is accompanied by increased levels of extracellular proteases that are capable of remodeling the extracellular matrix, as well as cleaving and activating growth factors and receptors that are involved in pro‐cancerous signaling pathways. Several members of the type II transmembrane serine protease (TTSP) family have been shown to play critical roles in cancer progression, however, the expression or function of the TTSP Human Airway Trypsin‐like protease (HAT) in carcinogenesis has not been examined. In the present study we aimed to determine the expression of HAT during squamous cell carcinogenesis. HAT transcript is present in several tissues containing stratified squamous epithelium and decreased expression is observed in carcinomas. We determined that HAT protein is consistently expressed on the cell surface in suprabasal/apical layers of squamous cells in healthy cervical and esophageal epithelia. To assess whether HAT protein is differentially expressed in normal tissue versus tissue in different stages of carcinogenesis, we performed a comprehensive immunohistochemical analysis of HAT protein expression levels and localization in arrays of paraffin embedded human cervical and esophageal carcinomas compared to the corresponding normal tissue. We found that HAT protein is expressed in the non‐proliferating, differentiated cellular strata and is lost during the dedifferentiation of epithelial cells, a hallmark of squamous cell carcinogenesis. Thus, HAT expression may potentially be useful as a marker for clinical grading and assessment of patient prognosis in squamous cell carcinomas. This article is protected by copyright. All rights reserved

  August 22, 2015   doi: 10.1002/jcp.25173   open full text
• Defining Plasma MicroRNAs Associated With Cognitive Impairment in Hiv‐Infected Patients.
  Ferdous Kadri, Andrea LaPlante, Mariacristina De Luca, Lisa Doyle, Cruz Velasco‐Gonzalez, Jonathan R. Patterson, Patricia E. Molina, Steve Nelson, Arnold Zea, Christopher H. Parsons, Francesca Peruzzi.
  Journal of Cellular Physiology. August 18, 2015
  Human Immunodeficiency Virus (HIV)‐infected individuals are at increased risk for developing neurocognitive disorders and depression. These conditions collectively affect more than 50% of people living with HIV/AIDS and adversely impact adherence to HIV therapy. Thus, identification of early markers of neurocognitive impairment could lead to interventions that improve psychosocial functioning and slow or reverse disease progression through improved treatment adherence. Evidence has accumulated for the role and function of microRNAs in normal and pathological conditions. We have optimized a protocol to profile microRNAs in body fluids. Using this methodology, we have profiled plasma microRNA expression for 30 age‐matched, HIV‐infected (HIV+) patients and identified highly sensitive and specific microRNA signatures distinguishing HIV+ patients with cognitive impairment from those without cognitive impairment. These results justify follow‐on studies to determine whether plasma microRNA signatures can be used as a screening or prognostic tool for HIV+ patients with neurocognitive impairment. This article is protected by copyright. All rights reserved

  August 18, 2015   doi: 10.1002/jcp.25131   open full text
• The Histone‐Deacetylase‐Inhibitor Suberoylanilide Hydroxamic Acid Promotes Dental Pulp Repair Mechanisms Through Modulation of Matrix Metalloproteinase‐13 Activity.
  Henry F. Duncan, Anthony J. Smith, Garry J. P. Fleming, Nicola C. Partridge, Emi Shimizu, Gary P. Moran, Paul R. Cooper.
  Journal of Cellular Physiology. August 11, 2015
  Direct application of histone‐deacetylase‐inhibitors (HDACis) to dental pulp cells (DPCs) induces chromatin changes, promoting gene expression and cellular‐reparative events. We have previously demonstrated that HDACis (Valproic acid, Trichostatin A) increase mineralization in dental papillae‐derived cell‐lines and primary DPCs by stimulation of dentinogenic gene expression. Here, we investigated novel genes regulated by the HDACi, suberoylanilide hydroxamic acid (SAHA), to identify new pathways contributing to DPC differentiation. SAHA significantly compromised DPC viability only at relatively high concentrations (5 μM); while low concentrations (1 μM) SAHA did not increase apoptosis. HDACi‐exposure for 24 h induced mineralization‐per‐cell dose‐dependently after 2 weeks; however, constant 14d SAHA‐exposure inhibited mineralization. Microarray analysis (24 h and 14d) of SAHA exposed cultures highlighted that 764 transcripts showed a significant >2.0‐fold change at 24 h, which reduced to 36 genes at 14d. 59% of genes were down‐regulated at 24 h and 36% at 14d, respectively. Pathway analysis indicated SAHA increased expression of members of the matrix metalloproteinase (MMP) family. Furthermore, SAHA‐supplementation increased MMP‐13 protein expression (7d, 14 d) and enzyme activity (48 h, 14d). Selective MMP‐13‐inhibition (MMP‐13i) dose‐dependently accelerated mineralization in both SAHA‐treated and non‐treated cultures. MMP‐13i‐supplementation promoted expression of several mineralization‐associated markers, however, HDACi‐induced cell migration and wound healing were impaired. Data demonstrate that short‐term low‐dose SAHA‐exposure promotes mineralization in DPCs by modulating gene pathways and tissue proteases. MMP‐13i further increased mineralization‐associated events, but decreased HDACi cell migration indicating a specific role for MMP‐13 in pulpal repair processes. Pharmacological inhibition of HDAC and MMP may provide novel insights into pulpal repair processes with significant translational benefit. This article is protected by copyright. All rights reserved

  August 11, 2015   doi: 10.1002/jcp.25128   open full text
• Autophagy‐Induced Apoptosis in Lung Cancer Cells By a Novel Digitoxin Analog.
  Yogesh M. Kulkarni, Vivek Kaushik, Neelam Azad, Clayton Wright, Yon Rojanasakul, George O'Doherty, Anand Krishnan V. Iyer.
  Journal of Cellular Physiology. August 11, 2015
  We have synthesized a novel derivative of Digitoxin, termed “MonoD”, which demonstrates cytotoxic effects in lung cancer cells with much higher potency as compared to Digitoxin. Our data show that within one hour of MonoD treatment, H460 cells showed increased oxidative stress, increased formation of autophagic vacuoles and increased expression of pro‐autophagic markers Beclin‐1 and LC3‐II. Cells pretreated with MnTBAP, a superoxide scavenger not only lowered superoxide production, but also had lower levels of LC3‐II and Beclin‐1. Prolonged treatment with MonoD induced apoptosis in lung cancer cells. We investigated MonoD‐dependent regulation of Akt and Bcl2, proteins that are known regulators of both autophagy and apoptosis. Molecular and pharmacologic inhibitors of Bcl2 and Akt, when combined with MonoD, led to higher expression of LC3‐II and Beclin‐1 as compared to MonoD alone, suggesting a repressive effect for these proteins in MonoD‐dependent autophagy. Pretreatment of cells with an autophagy inhibitor repressed the apoptotic potential of MonoD, confirming that early autophagic flux is important to drive apoptosis. Therapeutic entities such as MonoD that target multiple pathways such as autophagy and apoptosis may prove advantageous over current therapies that have unimodal basis for action and may drive sustained tumor regression, which is highly desirable. This article is protected by copyright. All rights reserved

  August 11, 2015   doi: 10.1002/jcp.25129   open full text
• CD44 Influences Fibroblast Behaviors Via Modulation of Cell‐Cell and Cell‐Matrix Interactions, Affecting Survivin and Hippo Pathways.
  Masayuki Tsuneki, Joseph A. Madri.
  Journal of Cellular Physiology. August 06, 2015
  CD44 has been studied in a wide variety of cell types, in a diverse array of cell behaviors and in a diverse range of signaling pathways. We now document a role for CD44 in mediating fibroblast behaviors via regulation of N‐cadherin, extracellular matrix expression, Survivin and the Hippo pathway. Here we report our findings on the roles of CD44 in modulating proliferation, apoptosis, migration and invasion of murine wild‐type (WT‐FB) and CD44 knockout dermal fibroblasts (CD44KO‐FB). As we have documented in microvascular endothelial cells lacking CD44, we found persistent increased proliferation, reduced activation of cleaved caspase 3, increased initial attachment, but decreased strength of cell attachment in high cell density, post confluent CD44KO‐FB cultures. Additionally, we found that siRNA knock‐down of CD44 mimicked the behaviors of CD44KO‐FB, restoring the decreases in N‐cadherin, collagen type I, fibronectin, Survivin, nuclear fractions of YAP and phospho‐YAP and decreased levels of cleaved caspase 3 to the levels observed in CD44KO‐FB. Interestingly, plating CD44KO‐FB on collagen type I or fibronectin resulted in significant decreases in secondary proliferation rates compared to plating cells on non‐coated dishes, consistent with increased cell adhesion compared to their effects on WT‐FB. Lastly, siRNA knockdown of CD44 in WT‐FB resulted in increased fibroblast migration compared to WT‐FB, albeit at reduced rates compared to CD44KO‐FB. These results are consistent with CD44's pivotal role in modulating several diverse behaviors important for adhesion, proliferation, apoptosis, migration and invasion during development, growth, repair, maintenance and regression of a wide variety of mesenchymal tissues. This article is protected by copyright. All rights reserved

  August 06, 2015   doi: 10.1002/jcp.25123   open full text
• Nuclear Translocation of p65 is Controlled by Sec6 Via the Degradation of IκBα.
  Toshiaki Tanaka, Mitsuyoshi Iino.
  Journal of Cellular Physiology. August 06, 2015
  Nuclear factor‐κB (NF‐κB) is an inducible transcription factor that mediates immune and inflammatory responses. NF‐κB pathways are also involved in cell adhesion, differentiation, proliferation, autophagy, senescence, and protection against apoptosis. The deregulation of NF‐κB activity is found in a number of disease states, including cancer, arthritis, chronic inflammation, asthma, neurodegenerative diseases, and heart disease. The 90 kDa ribosomal S6 kinase (p90RSK) family, which is serine/threonine kinases, is phosphorylated by extracellular signal‐regulated kinase1/2 (ERK1/2) and is related to NF‐κB pathways. Our previous studies revealed that Sec6, a component of the exocyst complex, plays specific roles in cell‐cell adhesion and cell cycle arrest. However, the mechanism by which Sec6 regulates the NF‐κB signaling pathway is unknown. We demonstrated that Sec6 knockdown inhibited the degradation of IκBα and delayed the nucleus‐cytoplasm translocation of p65 in HeLa cells transfected with Sec6 siRNAs after treatment with tumor necrosis factor alpha (TNF‐α). Furthermore, the binding of p65 and cAMP response element binding protein (CREB) binding protein (CBP) or p300 decreased and NF‐κB related genes which were inhibitors of NF‐κB alpha (IκBα), A20, B cell lymphoma protein 2 (Bcl‐2), and monocyte chemoattractant protein‐1 (MCP‐1) were low in cells transfected with Sec6 siRNAs in response to TNF‐α stimulation. Sec6 knockdown decreased the expression of p90RSKs and the phosphorylation of ERK or p90RSK1 at Ser380 or IκBα at Ser32. The present study suggests that Sec6 regulates NF‐κB transcriptional activity via the control of the phosphorylation of IκBα, p90RSK1, and ERK. This article is protected by copyright. All rights reserved

  August 06, 2015   doi: 10.1002/jcp.25122   open full text
• Retinoblastoma Protein Knockdown Favors Oxidative Metabolism and Glucose and Fatty Acid Disposal in Muscle Cells.
  Petar D. Petrov, Joan Ribot, Isabel C. López‐Mejía, Lluís Fajas, Andreu Palou, M. Luisa Bonet.
  Journal of Cellular Physiology. August 04, 2015
  Deficiency in the retinoblastoma protein (Rb) favors leanness and a healthy metabolic profile in mice largely attributed to activation of oxidative metabolism in white and brown adipose tissues. Less is known about Rb modulation of skeletal muscle metabolism. This was studied here by transiently knocking down Rb expression in differentiated C2C12 myotubes using small interfering RNAs. Compared with control cells transfected with non‐targeting RNAs, myotubes silenced for Rb (by 80–90%) had increased expression of genes related to fatty acid uptake and oxidation such as Cd36 and Cpt1b (by 61% and 42%, respectively), increased Mitofusin 2 protein content (∼2.5‐fold increase), increased mitochondrial to nuclear DNA ratio (by 48%), increased oxygen consumption (by 65%) and decreased intracellular lipid accumulation. Rb silenced myotubes also displayed up‐regulated levels of glucose transporter type 4 expression (∼5‐fold increase), increased basal glucose uptake, and enhanced insulin‐induced Akt phosphorylation. Interestingly, exercise in mice led to increased Rb phosphorylation (inactivation) in skeletal muscle as evidenced by immunohistochemistry analysis. In conclusion, the silencing of Rb enhances mitochondrial oxidative metabolism and fatty acid and glucose disposal in skeletal myotubes, and changes in Rb status may contribute to muscle physiological adaptation to exercise. This article is protected by copyright. All rights reserved

  August 04, 2015   doi: 10.1002/jcp.25121   open full text
• Fucoidan Suppresses the Growth of Human Acute Promyelocytic Leukemia Cells In Vitro and In Vivo.
  Farzaneh Atashrazm, Ray M Lowenthal, Gregory M Woods, Adele F Holloway, Samuel S Karpiniec, Joanne L Dickinson.
  Journal of Cellular Physiology. August 03, 2015
  Fucoidan, a natural component of seaweeds, is reported to have immunomodulatory and anti‐tumor effects. The mechanisms underpinning these activities remain poorly understood. In this study, the cytotoxicity and anti‐tumor activities of fucoidan were investigated in acute myeloid leukemia (AML) cells. The human AML cell lines NB4, KG1a, HL60 and K562 were treated with fucoidan and cell cycle, cell proliferation and expression of apoptotic pathways molecules were analyzed. Fucoidan suppressed the proliferation and induced apoptosis through the intrinsic and extrinsic pathways in the acute promyelocytic leukemia (APL) cell lines NB4 and HL60, but not in KG1a and K562 cells. In NB4 cells, apoptosis was caspase‐dependent as it was significantly attenuated by pre‐treatment with a pan‐caspase inhibitor. P21/WAF1/CIP1 was significantly up‐regulated leading to cell cycle arrest. Fucoidan decreased the activation of ERK1/2 and down‐regulated the activation of AKT through hypo‐phosphorylation of Thr(308) residue but not Ser(473). In vivo, a xenograft model using the NB4 cells was employed. Mice were fed with fucoidan and tumor growth was measured following inoculation with NB4 cells. Subsequently, splenic natural killer (NK) cell cytotoxic activity was also examined. Oral doses of fucoidan significantly delayed tumor growth in the xenograft model and increased cytolytic activity of NK cells. Taken together, these data suggest that the selective inhibitory effect of fucoidan on APL cells and its protective effect against APL development in mice warrant further investigation of fucoidan as a useful agent in treatment of certain types of leukemia. This article is protected by copyright. All rights reserved

  August 03, 2015   doi: 10.1002/jcp.25119   open full text
• Impaired Hematopoiesis and Disrupted Monocyte/Macrophage Homeostasis in Mucopolysaccharidosis Type I Mice.
  Gustavo Monteiro Viana, Marcus Vinícius Buri, Edgar Julian Paredes‐Gamero, Ana Maria Martins, Vânia D'Almeida.
  Journal of Cellular Physiology. August 03, 2015
  Mucopolysaccharidosis type I (MPS I) is a rare autosomal recessive disease caused by alpha‐L‐iduronidase deficiency in which heparan and dermatan sulfate degradation is compromised. Besides primary lysosomal glycosaminoglycan accumulation, further changes in cellular functions have also been described in several murine MPS models. Herein, we evaluated alterations in hematopoiesis and its implications on the production of mature progeny in a MPS I murine model. Despite the significant increase in hematopoietic stem cells, a reduction in common myeloid progenitors and granulocyte‐macrophage progenitor cells was observed in Idua ‐/‐ mice bone marrow. Furthermore, no alterations in number, viability nor activation of cell death mechanisms were observed in Idua ‐/‐ mice mature macrophages but they presented higher sensitivity to apoptotic induction after staurosporine treatment. In addition, changes in Ca2+ signaling and a reduction in phagocytosis ability were also found. In summary, our results revealed significant intracellular changes in mature Idua ‐/‐ macrophages related to alterations in Idua ‐/‐ mice hematopoiesis, revealing a disruption in cell homeostasis. These results provide new insights into physiopathology of MPS I. This article is protected by copyright. All rights reserved

  August 03, 2015   doi: 10.1002/jcp.25120   open full text
• Hypoxia Promotes the Inflammatory Response and Stemness Features in Visceral Fat Stem Cells from Obese Subjects.
  Elisa Petrangeli, Giuseppe Coroniti, Anna T. Brini, Laura de Girolamo, Deborah Stanco, Stefania Niada, Gianfranco Silecchia, Emanuela Morgante, Carla Lubrano, Matteo A. Russo, Luisa Salvatori.
  Journal of Cellular Physiology. July 30, 2015
  Low‐grade chronic inflammation is a salient feature of obesity and many associated disorders. This condition frequently occurs in central obesity and is connected to alterations of the visceral adipose tissue (AT) microenvironment. Understanding how obesity is related to inflammation may allow the development of therapeutics aimed at improving metabolic parameters in obese patients. To achieve this aim, we compared the features of 2 subpopulations of adipose‐derived stem cells (ASC) isolated from both subcutaneous and visceral AT of obese patients with the features of 2 subpopulations of ASC from the same isolation sites of non‐obese individuals. In particular, the behavior of ASC of obese vs non‐obese subjects during hypoxia, which occurs in obese AT and is an inducer of the inflammatory response, was evaluated. Obesity deeply influenced ASC from visceral AT (obV‐ASC); these cells appeared to exhibit clearly distinguishable morphology and ultrastructure as well as reduced proliferation, clonogenicity and expression of stemness, differentiation and inflammation‐related genes. These cells also exhibited a deregulated response to hypoxia, which induced strong tissue‐specific NF‐kB activation and an NF‐kB‐mediated increase in inflammatory and fibrogenic responses. Moreover, obV‐ASC, which showed a less stem‐like phenotype, recovered stemness features after hypoxia. Our findings demonstrated the peculiar behavior of obV‐ASC, their influence on the obese visceral AT microenvironment and the therapeutic potential of NF‐kB inhibitors. These novel findings suggest that the deregulated hyper‐responsiveness to hypoxic stimulus of ASC from visceral AT of obese subjects may contribute via paracrine mechanisms to low‐grade chronic inflammation, which has been implicated in obesity‐related morbidity. This article is protected by copyright. All rights reserved

  July 30, 2015   doi: 10.1002/jcp.25113   open full text
• Regulation of Spermatogenic Cell T‐Type Ca2+ Currents by Zn2+: Implications in Male Reproductive Physiology.
  Ignacio López‐González, Claudia L. Treviño, Alberto Darszon.
  Journal of Cellular Physiology. July 29, 2015
  Zn2+ is a trace metal which is important for spermatogenesis progression; its deficiency causes atrophy or malignant growth of the testis. Although testis, epididymis and prostate contain high Zn2+ concentrations, the molecular entities which are modulated by this metal are still under study. Interestingly, spermatogenic cells mainly express CaV3.2‐encoded T‐type Ca2+ currents (ICaT) which are positively or negatively modulated by Zn2+ in other tissues. To explore whether ICaT could be regulated by Zn2+ and albumin, its main physiological carrier, we performed whole cell electrophysiological recordings of spermatogenic cell ICaT in the absence or presence of different Zn2+ concentrations. Zn2+ decreased ICaT in a concentration‐dependent manner (IC50= 2 μM) and this inhibition could only be completely removed in presence of albumin. Differently to previous reports, ICaT did not show a tonic inhibition by Zn2+. Further analysis showed that Zn2+ did not affect the voltage dependency or the kinetics of current activation, but right shifted the steady‐state inactivation curve and slowed inactivation and deactivation kinetics. Recovery from inactivation was also altered. However, these apparent alterations in gating properties are not enough to explain the strong ICaT reduction. Using non‐stationary fluctuation analysis, we found that Zn2+ mainly reduced the number of available Ca2+ channels without changing the single channel current amplitude. ICaT modulation by Zn2+ could be relevant for spontaneous Ca2+ oscillations during spermatogenesis and in pathophysiological conditions such as diabetes. This article is protected by copyright. All rights reserved

  July 29, 2015   doi: 10.1002/jcp.25112   open full text
• Failure to Target RANKL Signaling Through p38‐MAPK Results in Defective Osteoclastogenesis in the Microphthalmia Cloudy‐eyed Mutant.
  Heather A. Carey, Agnieszka Bronisz, Jennifer Cabrera, Blake E. Hildreth, Maria Cuitiño, Qi Fu, Asrar Ahmad, Ramiro E. Toribio, Michael C. Ostrowski, Sudarshana M. Sharma.
  Journal of Cellular Physiology. July 27, 2015
  The microphthalmia‐associated transcription factor (MITF) is a basic helix‐loop‐helix leucine zipper family factor that is essential for terminal osteoclast differentiation. Previous work demonstrates that phosphorylation of MITF by p38 MAPK downstream of receptor activator of NFκB ligand (RANKL) signaling is necessary for MITF activation in osteoclasts. The spontaneous Mitf cloudy eyed (ce) allele results in production of a truncated MITF protein that lacks the leucine zipper and C‐terminal end. Here we show that the Mitfce allele leads to a dense bone phenotype in neonatal mice due to defective osteoclast differentiation. In response to RANKL stimulation, in vitro osteoclast differentiation was impaired in myeloid precursors derived from neonatal or adult Mitfce/ce mice. The loss of the leucine zipper domain in Mitfce/ce mice does not interfere with the recruitment of MITF/PU.1 complexes to target promoters. Further, we have mapped the p38 MAPK docking site within the region deleted in Mitfce. This interaction is necessary for the phosphorylation of MITF by p38 MAPK. Site‐directed mutations in the docking site interfered with the interaction between MITF and its co‐factors FUS and BRG1. MITF‐ce fails to recruit FUS and BRG1 to target genes, resulting in decreased expression of target genes and impaired osteoclast function. These results highlight the crucial role of signaling dependent MITF/ p38 MAPK interactions in osteoclast differentiation. This article is protected by copyright. All rights reserved

  July 27, 2015   doi: 10.1002/jcp.25108   open full text
• Sustained Inhibition of Proliferative Response After Transient FGF Stimulation is Mediated by Interleukin 1 Signaling.
  Ashleigh Poole, Doreen Kacer, Emily Cooper, Francesca Tarantini, Igor Prudovsky.
  Journal of Cellular Physiology. July 27, 2015
  Transient FGF stimulation of various cell types results in FGF memory – a sustained blockage of efficient proliferative response to FGF and other growth factors. FGF memory establishment requires HDAC activity, indicating its epigenetic character. FGF treatment stimulates proinflammatory NFκB signaling, which is also critical for FGF memory formation. The search for FGF‐induced mediators of FGF memory revealed that FGF stimulates HDAC‐dependent expression of the inflammatory cytokine IL1α. Similarly to FGF, transient cell treatment with recombinant IL1α inhibits the proliferative response to further FGF and EGF stimulation, but does not prevent FGF receptor‐mediated signaling. Interestingly, like cells pretreated with FGF1, cells pretreated with IL1α exhibit enhanced restructuring of actin cytoskeleton and increased migration in response to FGF stimulation. IRAP, a specific inhibitor of IL1 receptor, and a neutralizing anti‐IL1α antibody prevent the formation of FGF memory and rescue an efficient proliferative response to FGF restimulation. A similar effect results following treatment with the anti‐inflammatory agents aspirin and dexamethasone. Thus, FGF memory is mediated by proinflammatory IL1 signaling. It may play a role in the limitation of proliferative response to tissue damage and prevention of wound‐induced hyperplasia. This article is protected by copyright. All rights reserved

  July 27, 2015   doi: 10.1002/jcp.25111   open full text
• Leptin Effect on Acetylation and Phosphorylation of PGC1α in Muscle Cells Associated With AMPK and Akt Activation in High‐Glucose Medium.
  Francisco García‐Carrizo, Yuriy Nozhenko, Andreu Palou, Ana M. Rodríguez.
  Journal of Cellular Physiology. July 27, 2015
  Leptin is crucial in energy metabolism, including muscle regulation. Peroxisome proliferator activated receptor gamma co‐activator 1α (PGC1α) orchestrates energy metabolism and is tightly controlled by post‐translational covalent modifications such as phosphorylation and acetylation. We aimed to further the knowledge of PGC1α control by leptin (at physiological levels) in muscle cells by time‐sequentially analysing the activation of AMP activated protein kinase (AMPK), p38 mitogen‐activated protein kinase (p38 MAPK) and Akt (Protein kinase B) – all known to phosphorylate PGC1α and to be involved in the regulation of its acetylation status – in C2C12 myotubes placed in a high‐glucose serum‐free medium. We also studied the protein levels of PGC1α, Sirtuin 1, adiponectin, COX IV, mitofusin 2 (Mfn2) and pyruvate dehydrogenase kinase 4 (PDK4). Our main findings suggest an important role of leptin regulating AMPK and Akt phosphorylation, mitofusin 2 induction and PGC1α acetylation status, with the novelty that the latter in transitorily increased in response to leptin, an effect dependent, at least in part, on AMPK regulation. These post‐translational reversible changes in PGC1α in response to leptin, especially the increase in acetylation status, may be related to the physiological role of the hormone in modulating muscle cell response to the physiological/nutritional status. This article is protected by copyright. All rights reserved

  July 27, 2015   doi: 10.1002/jcp.25109   open full text
• BMP‐2 Induced Expression of PLCβ1 That Is a Positive Regulator of Osteoblast Differentiation.
  Giulia Ramazzotti, Alberto Bavelloni, William Blalock, Manuela Piazzi, Lucio Cocco, Irene Faenza.
  Journal of Cellular Physiology. July 27, 2015
  Bone morphogenetic protein 2 (BMP‐2) is a critical growth factor that directs osteoblast differentiation and bone formation. Phosphoinositide‐phospholipase Cβ 1 (PLCβ1) plays a crucial role in the initiation of the genetic program responsible for muscle differentiation. Differentiation of C2C12 mouse myoblasts in response to insulin stimulation is characterized by a marked increase in nuclear PLCβ1. Here, the function of PLCβ1 in the osteogenic differentiation was investigated. Briefly, in C2C12 cells treated with BMP‐2 we assist to a remarkable increase in PLCβ1 protein and mRNA expression. The data regarding the influence on differentiation demonstrated that PLCβ1 promotes osteogenic differentiation by up‐regulating alkaline phosphatase (ALP). Moreover, PLCβ1 is present in the nuclear compartment of these cells and overexpression of a cytosolic‐PLCβ1mutant (cyt‐PLCβ1), which lacks a nuclear localization sequence, prevented the differentiation of C2C12 cells into osteocytes. Recent evidence indicates that miRNAs act as important post transcriptional regulators in a large number of processes, including osteoblast differentiation. Since miR‐214 is a regulator of Osterix (Osx) which is an osteoblast‐specific transcription factor that is needful for osteoblast differentiation and bone formation, we further investigated whether PLCβ1 could be a potential target of miR‐214 in the control of osteogenic differentiation by gain‐ and loss‐ of function experiment. The results indicated that inhibition of miR‐214 in C2C12 cells significantly enhances the protein level of PLCβ1 and promotes C2C12 BMP‐2‐induced osteogenesis by targeting PLCβ1. This article is protected by copyright. All rights reserved

  July 27, 2015   doi: 10.1002/jcp.25107   open full text
• Reciprocal Control of Osteogenic and Adipogenic Differentiation by ERK/MAP Kinase Phosphorylation of Runx2 and PPARγ Transcription Factors.
  Chunxi Ge, William P. Cawthorn, Yan Li, Guisheng Zhao, Ormond A. MacDougald, Renny T. Franceschi.
  Journal of Cellular Physiology. July 23, 2015
  In many skeletal diseases, including osteoporosis and disuse osteopenia, defective osteoblast differentiation is associated with increased marrow adipogenesis. The relative activity of two transcription factors, RUNX2 and PPARγ, controls whether a mesenchymal cell will differentiate into an osteoblast or adipocyte. Herein we show that the ERK/MAP kinase pathway, an important mediator of mechanical and hormonal signals in bone, stimulates osteoblastogenesis and inhibits adipogenesis via phosphorylation of RUNX2 and PPARγ. Induction of osteoblastogenesis in ST2 mesenchymal cells was associated with increased MAPK activity and RUNX2 phosphorylation. Under these conditions PPARγ phosphorylation also increased, but adipogenesis was inhibited. In contrast, during adipogenesis MAPK activity and phosphorylation of both transcription factors was reduced. RUNX2 phosphorylation and transcriptional activity were directly stimulated by MAPK, a response requiring phosphorylation at S301 and S319. MAPK also inhibited PPARγ‐dependent transcription via S112 phosphorylation. Stimulation of MAPK increased osteoblastogenesis and inhibited adipogenesis, while dominant‐negative suppression of activity had the opposite effect. In rescue experiments using Runx2‐/‐ mouse embryo fibroblasts (MEFs), wild type or, to a greater extent, phosphomimetic mutant RUNX2 (S301E,S319E) stimulated osteoblastogenesis while suppressing adipogenesis. In contrast, a phosphorylation‐deficient RUNX2 mutant (S301A,S319A) had reduced activity. Conversely, wild type or, to a greater extent, phosphorylation‐resistant S112A mutant PPARγ strongly stimulated adipogenesis and inhibited osteoblastogenesis in Pparg ‐/‐ MEFs, while S112E mutant PPARγ was less active. Competition between RUNX2 and PPARγ was also observed at the transcriptional level. Together, these studies highlight the importance of MAP kinase signaling and RUNX2/PPARγ phosphorylation in the control of osteoblast and adipocyte lineages. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25102   open full text
• Osteogenic Potential of Human Oral‐Periosteal Cells (PCs) Isolated from Different Oral Origin: An In Vitro Study.
  Gabriele Ceccarelli, Antonio Graziano, Laura Benedetti, Marcello Imbriani, Federica Romano, Francesco Ferrarotti, Mario Aimetti, Gabriella M. Cusella De Angelis.
  Journal of Cellular Physiology. July 23, 2015
  The periosteum is a specialized connective tissue containing multipotent stem cells capable of bone formation. In this study, we aimed at demonstrating that human oral periosteal cells derived from three different oral sites (upper vestibule, lower vestibule and hard palate) represent an innovative cell source for maxillo‐facial tissue engineering applications in terms of accessibility and self‐commitment towards osteogenic lineage. Periosteal cells (PCs) were isolated from patients with different ages (20‐30 yy, 40‐50 yy, 50‐60 yy); we then analyzed the in vitro proliferation capacity and the bone self‐commitment of cell clones culturing them without any osteogenic supplement to support their differentiation. We found that oral PCs, independently of their origin and age of patients, are mesenchymal stem cells with stem cell characteristics (clonogenical and proliferative activity) and that, even in absence of any osteogenic induction, they undertake the osteoblast lineage after 45 days of culture. These results suggest that oral periosteal cells could replace mesenchymal cells from bone marrow in oral tissue‐engineering applications. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25104   open full text
• CyclinD1 Down Regulation and Increased Apoptosis Are Common Features of Cohesinopathies.
  Grazia Fazio, Carles Gaston‐Massuet, Laura Rachele Bettini, Federica Graziola, Valeria Scagliotti, Anna Cereda, Luca Ferrari, Mara Mazzola, Gianni Cazzaniga, Antonio Giordano, Franco Cotelli, Gianfranco Bellipanni, Andrea Biondi, Angelo Selicorni, Anna Pistocchi, Valentina Massa.
  Journal of Cellular Physiology. July 23, 2015
  Genetic variants within components of the cohesin complex (NIPBL, SMC1A, SMC3, RAD21, PDS5, ESCO2, HDAC8) are believed to be responsible for a spectrum of human syndromes known as “cohesinopathies” that includes Cornelia de Lange Syndrome (CdLS). CdLS is a multiple malformation syndrome affecting almost any organ and causing severe developmental delay. Cohesinopathies seem to be caused by dysregulation of specific developmental pathways downstream of mutations in cohesin components. However, it is still unclear how mutations in different components of the cohesin complex affect the output of gene regulation. In this study, zebrafish embryos and SMC1A‐mutated patient‐derived fibroblasts were used to analyze abnormalities induced by SMC1A loss of function. We show that the knockdown of smc1a in zebrafish impairs neural development, increases apoptosis and specifically down‐regulates Ccnd1 levels. The same down‐regulation of cohesin targets is observed in SMC1A‐mutated patient fibroblasts. Previously, we have demonstrated that haploinsufficiency of NIPBL produces similar effects in zebrafish and in patients fibroblasts indicating a possible common feature for neurological defects and mental retardation in cohesinopathies. Interestingly, expression analysis of Smc1a and Nipbl in developing mouse embryos reveals a specific pattern in the hindbrain, suggesting a role for cohesin in neural development in vertebrates. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25106   open full text
• The PPARβ/δ Agonist GW0742 Induces Early Neuronal Maturation of Cortical Post‐Mitotic Neurons: Role of PPARβ/δ in Neuronal Maturation.
  E. Benedetti, S. Di Loreto, B. D'Angelo, L. Cristiano, M. d'Angelo, A. Antonosante, A. Fidoamore, Golini Raffaella, B. Cinque, MG Cifone, R Ippoliti, A. Giordano, A. Cimini.
  Journal of Cellular Physiology. July 23, 2015
  Increasing evidences support that signalling lipids participate in synaptic plasticity and cell survival, and that the lipid signalling is closely associated with neuronal differentiation, learning and memory and with pathologic events such as epilepsy and Alzheimer's disease. The Peroxisome Proliferator‐Activated Receptors (PPAR) are strongly involved in the fatty acid cell signalling, as many of the natural lypophylic compounds are PPAR ligands. We have previously shown that PPARβ/δ is the main isotype present in cortical neuron primary cultures and that during neuronal maturation, PPARβ/δ is gradually increased and activated. To get more insight into the molecular mechanism by which PPARβ/δ may be involved in neuronal maturation processes, in this work a specific PPARβ/δ agonist, GW0742 was used administered alone or in association with a specific PPARβ/δ antagonist, the GSK0660 and the parameters involved in neuronal differentiation and maturation were assayed. The data obtained demonstrated the strong involvement of PPARβ/δ in neuronal maturation, triggering the agonist an anticipation of neuronal differentiation and the antagonist abolishing the observed effects. These effects appear to be mediated by the activation of both BDNF canonical (BDNF7TrkB) pathway. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25103   open full text
• RhoA‐Mediated Functions in C3H10T1/2 Osteoprogenitors Are Substrate Topography‐Dependent.
  Yoichiro Ogino, Ruiwei Liang, Daniela B.S. Mendonça, Gustavo Mendonça, Masako Nagasawa, Kiyoshi Koyano, Lyndon F. Cooper.
  Journal of Cellular Physiology. July 23, 2015
  Surface topography broadly influences cellular responses. Adherent cell activities are regulated, in part, by RhoA, a member of the Rho‐family of GTPases. In this study, we evaluated the influence of surface topography on RhoA activity and associated cellular functions. The murine mesenchymal stem cell line C3H10T1/2 cells (osteoprogenitor cells) were cultured on titanium substrates with smooth topography (S), microtopography (M) and nanotopography (N) to evaluate the effect of surface topography on RhoA‐mediated functions (cell spreading, adhesion, migration and osteogenic differentiation). The influence of RhoA activity in the context of surface topography was also elucidated using RhoA pharmacologic inhibitor. Following adhesion, M and N adherent cells developed multiple projections, while S adherent cells had flattened and widespread morphology. RhoA inhibitor induced remarkable longer and thinner cytoplasmic projections on all surfaces. Cell adhesion and osteogenic differentiation was topography dependent with S< M and N surfaces. RhoA inhibition increased adhesion on S and M surfaces, but not N surfaces. Cell migration in a wound healing assay was greater on S versus M versus N surfaces and RhoA inhibitor increased S adherent cell migration, but not N adherent cell migration. RhoA inhibitor enhanced osteogenic differentiation in S adherent cells, but not M or N adherent cells. RhoA activity was surface topography roughness‐dependent (S < M, N). RhoA activity and ‐mediated functions are influenced by surface topography. Smooth surface adherent cells appear highly sensitive to RhoA function, while nano‐scale topography adherent cell may utilize alternative cellular signaling pathway(s) to influence adherent cellular functions regardless of RhoA activity. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25100   open full text
• IL‐6 Activates PI3K and PKCζ Signaling and Determines Cardiac Differentiation in Rat Embryonic H9c2 Cells.
  Maria Angela D'amico, Barbara Ghinassi, Pascal Izzicupo, Annalisa Di Ruscio, Angela Di Baldassarre.
  Journal of Cellular Physiology. July 23, 2015
  Introduction IL‐6 influences several biological processes, including cardiac stem cell and cardiomyocyte physiology. Although JAK‐STAT3 activation is the defining feature of IL‐6 signaling, signaling molecules such as PI3K, PKCs and ERK1/2 are also activated and elicit different responses. Moreover, most studies on the specific role of these signaling molecules focus on the adult heart, and few studies are available on the biological effects evoked by IL‐6 in embryonic cardiomyocytes. Aim The aim of this study was to clarify the biological response of embryonic heart derived cells to IL‐6 by analyzing the morphological modifications and the signaling cascades evoked by the cytokine in H9c2 cells. Results IL‐6 stimulation determined the terminal differentiation of H9c2 cells, as evidenced by the increased expression of cardiac transcription factors (NKX2.5 and GATA4), structural proteins (α‐myosin heavy chain and cardiac Troponin T) and the gap junction protein Connexin 43. This process was mediated by the rapid modulation of PI3K, Akt, PTEN and PKCζ phosphorylation levels. PI3K recruitment was an upstream event in the signaling cascade and when PI3K was inhibited, IL‐6 failed to modify PKCζ, PTEN and Akt phosphorylation. Blocking PKCζ activity affected only PTEN and Akt. Finally, the overexpression of a constitutively active form of PKCζ in H9c2 cells largely mimicked the morphological and molecular effects evoked by IL‐6. Conclusion This study demonstrated that IL‐6 induces the cardiac differentiation of H9c2 embryonic cells though a signaling cascade that involves PI3K, PTEN and PKCζ activities. This article is protected by copyright. All rights reserved

  July 23, 2015   doi: 10.1002/jcp.25101   open full text
• Agents Which Inhibit NF‐κB Signaling Block Spontaneous Contractile Activity and Negatively Influence Survival of Developing Myotubes.
  C. George Carlson, Lauren Stein, Elizabeth Dole, Ross M. Potter, David Bayless.
  Journal of Cellular Physiology. June 30, 2015
  Inhibiting the NF‐κB signaling pathway provides morphological and functional benefits for the mdx mouse, a model for Duchenne muscular dystrophy characterized by chronic elevations in the nuclear expression of p65, the transactivating component of the NF‐κB complex. The purpose of this study was to examine p65 expression in nondystrophic and mdx myotubes using confocal immunofluorescence, and determine whether inhibitors of the NF‐κB pathway alter myotube development. Primary cultures of nondystrophic and mdx myotubes had identical levels of nuclear and cytosolic p65 expression and exhibited equivalent responses to TNF‐α, thus excluding the hypothesis that the lack of dystrophin is sufficient to induce increases in NF‐κB signaling. The NF‐κB inhibitors pyrrolidine dithiocarbamate (PDTC) and sulfasalazine decreased spontaneous contractile activity and reduced myotube viability in a dose‐ and time‐dependent manner. Similarly, a vivo‐morpholino designed to block translation of murine p65 (m‐p65tb‐vivomorph1) rapidly abolished spontaneous contractile activity, reduced p65 expression measured by confocal immunofluorescence, and induced cell death in primary cultures of nondystrophic and mdx myotubes. Similar effects on p65 immunofluorescence and cell viability were observed following m‐p65tb‐vivomorph1 exposure to spontaneously inactive C2C12 myotubes, while exposure to a control scrambled vivo morpholino had no effect. These results indicate a direct role of the NF‐κB pathway in myotube development and identify a potential therapeutic limitation to the use of NF‐κB inhibitors in treating Duchenne and related muscular dystrophies. This article is protected by copyright. All rights reserved

  June 30, 2015   doi: 10.1002/jcp.25085   open full text
• Electrospun Type 1 Collagen Matrices Using a Novel Benign Solvent for Cardiac Tissue Engineering.
  Alan M. Punnoose, Anuradha Elamparithi, Sarah Kuruvilla.
  Journal of Cellular Physiology. May 12, 2015
  --- - |2+ Electrospinning is a well‐established technique that uses a high electric field to fabricate ultra fine fibrous scaffolds from both natural and synthetic polymers to mimic the cellular microenvironment. Collagen is one of the most preferred biopolymers due to its biocompatibility and widespread occurrence in nature. Electrospinning of Collagen alone has been reported with fluoroalcohols such as Hexafluoroisopropanol (HFIP) and Trifluoroethanol (TFE), which are toxic to the environment. In this study we describe the use of a novel benign binary solvent to generate nanofibers of Collagen type 1, which is non‐toxic and economical. Transmission electron microscopy (TEM) analysis revealed the characteristic feature of native collagen namely the 67 nm banding pattern, confirming that the triple helical structure was maintained. Scanning Electron Microscopy (SEM) analysis showed the fiber diameters to be in the 200‐800 nm range. Biocompatibility of the three dimensional (3D) scaffolds was established by MTT assays using skeletal myoblasts and Confocal Microscopic analysis of immunofluorescent stained sections for muscle specific markers such as Desmin and Actin. Primary neonatal rat ventricular cardiomyocytes seeded onto the scaffolds were able to maintain their contractile function for a period of 17 days. Our work provides evidence that Collagen 1 can be electrospun without combining with other polymers using a novel benign solvent and we are currently exploring the potential of this approach for cardiac and skeletal muscle tissue engineering. This article is protected by copyright. All rights reserved - Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-.
  

  May 12, 2015   doi: 10.1002/jcp.25035   open full text
• Protein Kinase C Is Involved in the Induction of ATP‐Binding Cassette Transporter A1 Expression by Liver X Receptor/Retinoid X Receptor Agonist in Human Macrophages.
  Etimad A. Huwait, Nishi N. Singh, Daryn R. Michael, Thomas S. Davies, Joe W. E. Moss, Dipak P. Ramji.
  Journal of Cellular Physiology. May 07, 2015
  --- - |2+ Abstract The transcription of the ATP‐binding cassette transporter A1 (ABCA1) gene, which plays a key anti‐atherogenic role, is known to be induced by agonists of liver X receptors (LXRs). LXRs form obligate heterodimers with retinoid X receptors (RXRs) and interact with their recognition sequences in the regulatory regions of key genes implicated in the control of cholesterol, fatty acid and glucose homeostasis. We have previously shown a novel role for c‐Jun N‐terminal kinase (JNK) and phosphoinositide 3‐kinase (PI3K) in the LXRs‐mediated induction of macrophage gene expression. Protein kinase C (PKC) is often found to regulate the action of nuclear receptors and cross talk between this kinase family and JNK and/or PI3K has been shown in several settings. We have therefore investigated a potential role for PKC in the action of LXR/RXR agonist 22‐(R)‐hydroxycholesterol (22‐(R)‐HC)/9‐cis‐retinoic acid (9cRA) in THP‐1 macrophages, including the induction of ABCA1 expression. The pan PKC inhibitor bisindoylmaleimide was found to attenuate the induction of ABCA1 protein expression, the activation of the JNK signaling pathway and the stimulation of activator protein‐1 (AP‐1) DNA binding activity in macrophages treated with 22‐(R)‐HC and 9cRA. The role of PKC in the action of these ligands was confirmed further by the use of more isotype‐specific inhibitors. These studies therefore reveal a potentially important role for PKC in the action of 22‐(R)‐HC and 9cRA in human macrophages. This article is protected by copyright. All rights reserved - Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-.
  

  May 07, 2015   doi: 10.1002/jcp.25157   open full text
• Dual Effect of Cyanidin on RANKL‐Induced Differentiation and Fusion of Osteoclasts.
  Ce Dou, Jianmei Li, Fei Kang, Zhen Cao, Xiaochao Yang, Hong Jiang, Bo Yang, Junyu Xiang, Jianzhong Xu, Shiwu Dong.
  Journal of Cellular Physiology. December 24, 2014
  Bone homeostasis is maintained by the balance between osteoblastic bone formation and osteoclastic bone resorption. Osteoclasts are multinucleated cells derived from hematopoietic stem cells (HSCs) or monocyte/macrophage progenitor cells and formed by osteoclasts precursors (OCPs) fusion. Cyanidin is an anthocyanin widely distributed in food diet with novel antioxidant activity. However, the effect of cyanidin on osteoclasts is still unknown. We investigated the effect of cyanidin on RANKL‐induced osteoclasts differentiation and cell fusion. The results showed that cyanidin had a dual effect on RANKL‐induced osteoclastogenesis. Lower dosage of cyanidin (< 1μg/ml) has a promoting effect on osteoclastogenesis while higher dosage of cyanidin (> 10μg/ml) has an inhibitory effect. Fusogenic genes like CD9, ATP6v0d2, DC‐STAMP, OC‐STAMP and osteoclasts related genes like NFATc1, mitf and c‐fos were all regulated by cyanidin consistent to its dual effect. Further exploration showed that low concentration of cyanidin could increase osteoclasts fusion whereas higher dosage of cyanidin lead to the increase of LXR‐β expression and activation which is suppressive to osteoclasts differentiaton. All these results showed that cyanidin exhibits therapeutic potential in prevention of osteoclasts related bone disorders. This article is protected by copyright. All rights reserved

  December 24, 2014   doi: 10.1002/jcp.24916   open full text
• microRNA‐Mediated Survivin Control of Pluripotency.
  Kristina Kapinas, Heesun Kim, Matthew Mandeville, Lori A. Martin‐Buley, Carlo M. Croce, Jane B. Lian, Andre J. van Wijnen, Janet L. Stein, Dario C. Altieri, Gary S. Stein.
  Journal of Cellular Physiology. June 02, 2014
  Understanding the mechanisms that sustain pluripotency in human embryonic stem cells (hESCs) is an active area of research that may prove useful in regenerative medicine and will provide fundamental information relevant to development and cancer. hESCs and cancer cells share the unique ability to proliferate indefinitely and rapidly. Because the protein survivin is uniquely overexpressed in virtually all human cancers and in hESCs, we sought to investigate its role in supporting the distinctive capabilities of these cell types. Results presented here suggest that survivin contributes to the maintenance of pluripotency and that post‐transcriptional control of survivin isoform expression is selectively regulated by microRNAs. miR‐203 has been extensively studied in human tumors, but has not been characterized in hESCs. We show that miR‐203 expression and activity is consistent with the expression and subcellular localization of survivin isoforms that in turn modulate expression of the Oct4 and Nanog transcription factors to sustain pluripotency. This study contributes to understanding of the complex regulatory mechanisms that govern whether hESCs proliferate or commit to lineages. © 2014 Wiley Periodicals, Inc.

  June 02, 2014   doi: 10.1002/jcp.24681   open full text
• Pin1 Regulates Osteoclast Fusion through Suppression of the Master Regulator of Cell Fusion DC‐STAMP.
  Rabia Islam, Han‐Sol Bae, Won‐Joon Yoon, Kyung‐Mi Woo, Jeong‐Hwa Baek, Hong‐Hee Kim, Takafumi Uchida, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. June 02, 2014
  Cell fusion is a fundamental biological event that is essential for the development of multinucleated cells such as osteoclasts. Fusion failure leads to the accumulation of dense bone such as in osteopetrosis, demonstrating the importance of fusion in osteoclast maturity and bone remodeling. In a recent study, we reported that Pin1 plays a role in the regulation of bone formation and Runx2 regulation. In this study, we explored the role of Pin1 in osteoclast formation and bone resorption. Pin1 null mice have low bone mass and increased TRAP staining in histological sections of long bones, compared to Pin1 wild‐type mice. In vitro osteoclast forming assays with bone marrow‐derived monocyte/macrophage revealed that Pin1‐deficient osteoclasts are larger than wild‐type osteoclasts and have higher nuclei numbers, indicating greater extent of fusion. Pin1 deficiency also highly enhanced foreign body giant cell formation both in vitro and in vivo. Among the known fusion proteins, only DC‐STAMP was significantly increased in Pin1−/− osteoclasts. Immunohistochemistry showed that DC‐STAMP expression was also significantly increased in tibial metaphysis of Pin1 KO mice. We found that Pin1 binds and isomerizes DC‐STAMP and affects its expression levels and localization at the plasma membrane. Taken together, our data indicate that Pin1 is a determinant of bone mass through the regulation of the osteoclast fusion protein DC‐STAMP. The identification of Pin1 as a factor involved in cell fusion contributes to the understanding of osteoclast‐associated diseases, including osteoporosis, and opens new avenues for therapeutic targets. © 2014 Wiley Periodicals, Inc.

  June 02, 2014   doi: 10.1002/jcp.24679   open full text
• The Purα/Purβ Single‐Strand DNA‐Binding Proteins Attenuate Smooth‐Muscle Actin Gene Transactivation in Myofibroblasts.
  Seethalakshmi Hariharan, Robert J. Kelm, Arthur Roger Strauch.
  Journal of Cellular Physiology. May 23, 2014
  Expression of smooth muscle alpha‐actin (SMαA) is essential for myofibroblast‐mediated wound contraction following tissue injury. The Pur α/β and YB‐1 transcriptional repressors govern the DNA‐binding activity of serum response factor (SRF) and phosphorylated Smad3 (pSmad3) transcriptional activators during induction of SMαA gene expression in human pulmonary myofibroblasts. In quiescent fibroblasts, Pur α exhibited a novel function in enhancing stability of pre‐existing SRF complexes with SMαA core promoter DNA, whereas Pur β was more effective in disrupting SRF‐DNA interaction. Pur proteins were less efficient competitors of pre‐existing, core‐promoter complexes containing both SRF and pSmad3 in nuclear extracts from TGFβ1‐activated myofibroblasts. TGFβ1 signaling dissociated a SRF/Pur protein complex with concurrent formation of a transient pSmad3/MRTF‐A/Pur β complex during early phase myofibroblast differentiation. Pur β was replaced by Pur α in the pSmad3/MRTF‐A complex in mature myofibroblasts. Combining all three repressors potently inhibited SRF and pSmad3 binding to promoter DNA in quiescent fibroblasts and TGFβ1‐activated myofibroblasts, respectively. The results point to dynamic interplay between transcriptional activators and repressors in regulating SMαA gene output during myofibroblast differentiation. Therapeutic targeting of nucleoprotein complexes regulating the SMαA promoter may prevent excessive myofibroblast accumulation associated with chronic cardiopulmonary fibrosis and dysfunctional tissue remodeling. J. Cell. Physiol. 229: 1256–1271, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24564   open full text
• Cardioprotective Role of P38 MAPK During Myocardial Infarction Via Parallel Activation of α‐Crystallin B and Nrf2.
  Arkadeep Mitra, Aramita Ray, Ritwik Datta, Shantanu Sengupta, Sagartirtha Sarkar.
  Journal of Cellular Physiology. May 23, 2014
  Myocardial infarction (MI) is defined as cardiac cell death due to prolonged ischemia. Although necrotic cell death was considered to be solely responsible for myocyte death during MI, it was recently revealed that apoptosis also plays its part in this death process. Our laboratory has recently shown that endoplasmic reticulum (ER) stress‐induced apoptosis is the predominant route for apoptosis during MI and the conventional mitochondrial pathway is bypassed by activation of a small heat shock protein α‐crystallin B (CRYAB). Since CRYAB is a direct target of P38 mitogen‐activated protein kinase (MAPK) cascade, we were prompted to check the role of P38 MAPK in 20‐week‐old male Wister rats immediately after infarct formation. Interestingly, parallel activation of mitochondrial apoptotic pathway with an increase in ER stress‐induced apoptotic load was observed along with decreased activation of CRYAB and Nrf2 (a pro‐survival protein activated in response to ER stress) in MI rats treated with SB203580, a specific inhibitor of P38α and P38β compared to the MI alone. As a cumulative effect, this inhibitor treatment also resulted in significant increase in the levels of caspase3 activity and TUNEL positivity, the end point apoptotic markers. Furthermore, SB203580‐treated hypoxic adult cardiomyocytes showed formation of desmin aggregates which were previously associated with impaired cardiac function. Thus, this study shows for the first time the precise mechanism by which P38 MAPK plays a pro‐survival role and confers protection of cardiomyocytes, during infarct formation. J. Cell. Physiol. 229: 1272–1282, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24565   open full text
• Enzymatic and Regulatory Attributes of Trehalose‐6‐Phosphate Phosphatase from Candida utilis and its Role During Thermal Stress.
  Sagar Lahiri, Shakri Banerjee, Trina Dutta, Shinjinee Sengupta, Sandip Dey, Rusha Roy, Devlina Sengupta, Krishnananda Chattopadhyay, Anil K. Ghosh.
  Journal of Cellular Physiology. May 23, 2014
  Trehalose‐6‐phosphate phosphatase (TPP) catalyzes the final step in the biosynthesis of the anti‐stress sugar trehalose. An 82 kDa TPP enzyme was isolated from Candida utilis with 61% yield and 43‐fold purification. The protein sequence, determined by N‐terminal sequencing and MALDI‐TOF analysis, showed significant homology with known TPP sequences from related organisms. The full length gene sequence of TPP of C. utilis was identified using rapid amplification of cDNA ends‐PCR reaction (RACE‐PCR). The gene was cloned and expressed in Escherichia coli BL21. Recombinant TPP enzyme was isolated using affinity chromatography. CD spectroscopy and steady‐state fluorescence revealed that the structural and conformational aspects were identical in both native and recombinant forms. The biochemical properties of the two forms were also similar. Km was determined to be ~0.8 mM. Optimum temperature and pH were found to be 30 °C and 8.5, respectively. Activity was dependent on the presence of divalent cations and inhibited by metal chelators. Methylation‐mediated regulation of TPP enzyme and its effect on the overall survival of the organism under stress were investigated. The results indicated that enhancement of TPP activity by methylation at the Cysteine residues increased resistance of Candida cells against thermal stress. This work involves extensive investigations toward understanding the physico‐chemical properties of the first TPP enzyme from any yeast strain. The mechanism by which methylation regulates its activity has also been studied. A correlation between regulation of trehalose synthesis and survivability of the organism under thermal stress was established. J. Cell. Physiol. 229: 1245–1255, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24562   open full text
• Novel Adeno‐Associated Viral Vector Delivering the Utrophin Gene Regulator Jazz Counteracts Dystrophic Pathology in mdx Mice.
  Georgios Strimpakos, Nicoletta Corbi, Cinzia Pisani, Maria Grazia Di Certo, Annalisa Onori, Siro Luvisetto, Cinzia Severini, Francesca Gabanella, Lucia Monaco, Elisabetta Mattei, Claudio Passananti.
  Journal of Cellular Physiology. May 23, 2014
  Over‐expression of the dystrophin‐related gene utrophin represents a promising therapeutic strategy for Duchenne muscular dystrophy (DMD). The strategy is based on the ability of utrophin to functionally replace defective dystrophin. We developed the artificial zinc finger transcription factor “Jazz” that up‐regulates both the human and mouse utrophin promoter. We observed a significant recovery of muscle strength in dystrophic Jazz‐transgenic mdx mice. Here we demonstrate the efficacy of an experimental gene therapy based on the systemic delivery of Jazz gene in mdx mice by adeno‐associated virus (AAV). AAV serotype 8 was chosen on the basis of its high affinity for skeletal muscle. Muscle‐specific expression of the therapeutic Jazz gene was enhanced by adding the muscle α‐actin promoter to the AAV vector (mAAV). Injection of mAAV8‐Jazz viral preparations into mdx mice resulted in muscle‐specific Jazz expression coupled with up‐regulation of the utrophin gene. We show a significant recovery from the dystrophic phenotype in mAAV8‐Jazz‐treated mdx mice. Histological and physiological analysis revealed a reduction of fiber necrosis and inflammatory cell infiltration associated with functional recovery in muscle contractile force. The combination of ZF‐ATF technology with the AAV delivery can open a new avenue to obtain a therapeutic strategy for treatment of DMD. J. Cell. Physiol. 229: 1283–1291, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24567   open full text
• A Positive Role of MicroRNA‐15b on Regulation of Osteoblast Differentiation.
  S. Vimalraj, Nicola C. Partridge, N. Selvamurugan.
  Journal of Cellular Physiology. May 23, 2014
  Osteoblast differentiation is tightly regulated by several factors including microRNAs (miRNAs). In this paper, we report that pre‐mir‐15b is highly expressed in differentiated osteoblasts. The functional role of miR‐15b in osteoblast differentiation was determined using miR‐15b mimic/inhibitor and the expression of osteoblast differentiation marker genes such as alkaline phosphatase (ALP), type I collagen genes was decreased by miR‐15b inhibitor. Runx2, a bone specific transcription factor is generally required for expression of osteoblast differentiation marker genes and in response to miR‐15b inhibitor treatment, Runx2 mRNA expression was not changed; whereas its protein expression was decreased. Even though Smurf1 (SMAD specific E3 ubiquitin protein ligase 1), HDAC4 (histone deacetylase 4), Smad7, and Crim1 were found to be few of miR‐15b's putative target genes, there was increased expression of only Smurf1 gene at mRNA and protein levels by miR‐15b inhibitor. miR‐15b mimic treatment significantly increased and decreased expressions of Runx2 and Smurf1 proteins, respectively. We further identified that the Smurf1 3′UTR is directly targeted by miR‐15b using the luciferase reporter gene system. This is well documented that Smurf1 interacts with Runx2 and degrades it by proteasomal pathway. Hence, based on our results we suggest that miR‐15b promotes osteoblast differentiation by indirectly protecting Runx2 protein from Smurf1 mediated degradation. Thus, this study identified that miR‐15b can act as a positive regulator for osteoblast differentiation. J. Cell. Physiol. 229: 1236–1244, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24557   open full text
• SIRT1 Knockdown Promotes Neural Differentiation and Attenuates the Heat Shock Response.
  Diana J. Liu, David Hammer, Daniel Komlos, Kuang Yu Chen, Bonnie L. Firestein, Alice Y.‐C. Liu.
  Journal of Cellular Physiology. May 23, 2014
  Neurons have a limited capacity for heat shock protein (HSP) induction and are vulnerable to the pathogenic consequence of protein misfolding and aggregation as seen in age‐related neurodegenerative diseases. Sirtuin 1 (SIRT1), an NAD+‐dependent lysine deacetylase with important biological functions, has been shown to sustain the DNA‐binding state of HSF1 for HSP induction. Here we show that differentiation and maturation of embryonic cortical neurons and N2a neuroprogenitor cells is associated with decreases in SIRT1 expression and heat shock‐dependent induction of HSP70 protein. Tests of a pharmacological activator and an inhibitor of SIRT1 affirm the regulatory role of SIRT1 in HSP70 induction. Protein cross‐linking studies show that nuclear SIRT1 and HSF1 form a co‐migrating high molecular weight complex upon stress. The use of retroviral vectors to manipulate SIRT1 expression in N2a cells show that shRNA‐mediated knock down of SIRT1 causes spontaneous neurite outgrowth coincident with reduced growth rate and decreased induction of hsp70‐reporter gene, whereas SIRT1 over‐expression blocks the induced neural differentiation of N2a cells. Our results suggest that decreased SIRT1 expression is conducive to neuronal differentiation and this decrease contributes to the attenuated induction of HSPs in neurons. J. Cell. Physiol. 229: 1224–1235, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24556   open full text
• Human Placental Extract Mediated Inhibition of Proteinase K: Implications of Heparin and Glycoproteins in Wound Physiology.
  Kanika Sharma, Chaitali Mukherjee, Siddhartha Roy, Debashree De, Debasish Bhattacharyya.
  Journal of Cellular Physiology. May 23, 2014
  Efficient debridement of the wound bed following the removal of microbial load prevents its progression into a chronic wound. Bacterial infection and excessive proteolysis characterize impaired healing and therefore, their inhibition might restore the disturbed equilibrium in the healing process. Human placental extract exhibits reversible, non‐competitive inhibition towards Proteinase K, a microbial protease, by stabilizing it against auto‐digestion. Scattering and fluorescence studies followed by biochemical analysis indicated the involvement of a glycan moiety. Surface plasmon resonance demonstrated specific interaction of heparin with Proteinase K having Kd in μM range. Further, Proteinase K contains sequence motifs similar to other heparin‐binding proteins. Molecular docking revealed presence of clefts suitable for binding of heparin‐derived oligosaccharides. Comprehensive analysis of this inhibitory property of placental extract partly explains its efficacy in curing wounds with common bacterial infections. J. Cell. Physiol. 229: 1212–1223, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24555   open full text
• Adenosine Regulates the Proinflammatory Signaling Function of Thrombin in Endothelial Cells.
  Seyed Mahdi Hassanian, Peyman Dinarvand, Alireza R. Rezaie.
  Journal of Cellular Physiology. May 23, 2014
  The plasma level of the regulatory metabolite adenosine increases during the activation of coagulation and inflammation. Here we investigated the effect of adenosine on modulation of thrombin‐mediated proinflammatory responses in HUVECs. We found that adenosine inhibits the barrier‐disruptive effect of thrombin in HUVECs by a concentration‐dependent manner. Analysis of cell surface expression of adenosine receptors revealed that A2A and A2B are expressed at the highest level among the four receptor subtypes (A2B > A2A > A1 > A3) on HUVECs. The barrier‐protective effect of adenosine in response to thrombin was recapitulated by the A2A specific agonist, CGS 21680, and abrogated both by the siRNA knockdown of the A2A receptor and by the A2A‐specific antagonists, ZM‐241385 and SCH‐58261. The thrombin‐induced RhoA activation and its membrane translocation were both inhibited by adenosine in a cAMP‐dependent manner, providing a molecular mechanism through which adenosine exerts a barrier‐protective function. Adenosine also inhibited thrombin‐mediated activation of NF‐κB and decreased adhesion of monocytic THP‐1 cells to stimulated HUVECs via down‐regulation of expression of cell surface adhesion molecules, VCAM‐1, ICAM‐1, and E‐selectin. Moreover, adenosine inhibited thrombin‐induced elevated expression of proinflammatory cytokines, IL‐6 and HMGB‐1; and chemokines, MCP‐1, CXCL‐1, and CXCL‐3. Taken together, these results suggest that adenosine may inhibit thrombin‐mediated proinflammatory signaling responses, thereby protecting the endothelium from injury during activation of coagulation and inflammation. J. Cell. Physiol. 229: 1292–1300, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24568   open full text
• Exosomal ATF3 RNA Attenuates Pro‐Inflammatory Gene MCP‐1 Transcription in Renal Ischemia‐Reperfusion.
  Hsi‐Hsien Chen, Pei‐Fang Lai, Yi‐Fan Lan, Ching‐Feng Cheng, Wen‐Bing Zhong, Yuh‐Feng Lin, Tzen‐Wen Chen, Heng Lin.
  Journal of Cellular Physiology. May 23, 2014
  Transcriptional repressor activating transcription factor 3 (ATF3) is induced by various stress stimuli, including inflammation‐induced renal injury. In addition, ATF3 also down‐regulates adhesion molecules like intercellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), and monocyte chemotactic protein‐1 (MCP‐1). However, the relation between up‐regulated ATF3 after renal ischemia/reperfusion (I/R) injury and MCP‐1 is not completely understood. In this study, we demonstrated that, in renal I/R induced inflammation, induction of adhesion molecules (interleukin‐6, P‐selectin, E‐selectin, ICAM, VCAM, and MCP‐1) was higher in ATF3‐knockout mice than in wild‐type animals. Molecular and biochemical analyses revealed that ATF3 binds to the ATF/CRE sites in the MCP‐1 promoter and inhibits the secretion of MCP‐1 from renal epithelial cells after I/R injury. Urinary exosome containing ATF3 RNA was 60‐fold higher in patients with acute kidney injury than in normal controls, but no difference in total urinary ATF3 RNA levels was found. In addition, in vitro study showed that exosome containing ATF3 RNA derived from epithelial cells also inhibits MCP‐1 expression in the epithelial cells and macrophage migration. Furthermore, direct administration of the epithelium‐derived exosomal ATF3 RNA attenuates I/R induced kidney injury. Together, our studies reveal a novel regulatory mechanism of MCP‐1 expression mediated by the exosomal ATF3 RNA under renal I/R insult and suggest a potential targeted therapy for I/R induced acute kidney injury. J. Cell. Physiol. 229: 1202–1211, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24554   open full text
• Tanshinone II A Inhibits Tat‐Induced HIV‐1 Transactivation Through Redox‐Regulated AMPK/Nampt Pathway.
  Hong‐Sheng Zhang, Xin‐Yu Chen, Tong‐Chao Wu, Feng‐Juan Zhang.
  Journal of Cellular Physiology. May 23, 2014
  Tat transactivating activity regulated by NAD+‐dependent histone deacetylase sirtuin1 (SIRT1) connects HIV transcription with the metabolic state of the cell. Nicotinamide phosphoribosyltransferase (Nampt) is a rate‐limiting enzyme in the mammalian NAD+ biosynthesis. Nampt, SIRT1, and AMPK were involved in inhibiting HIV‐1 transactivation through redox‐regulated pathway. Tanshinone II A is a main lipid‐soluble monomer derivative from the root of Salvia miltiorrhiza (Danshen) and tanshinone II A possess a variety of biological activities through redox signaling pathway. Here we investigated the effect of tanshinone II A on Tat‐induced HIV‐1 transactivation and the redox signaling pathway involved in it. As the results were shown, tanshinone II A reversed Tat‐induced reactive oxygen species (ROS) production and down‐regulation of glutathione (GSH) levels in TZM‐bl cells through up‐regulation of Nrf2 expression. Tanshinone II A reversed Tat‐induced inhibition of SIRT1 activity but not SIRT1 protein expression. Tanshinone II A reversed Tat‐induced inhibition of Nampt protein expression and depletion of NAD+ levels in TZM‐bl cells in a dose‐dependent manner. Tanshinone II A‐evoked Nampt expression was mediated by AMPK signaling pathway. Tanshinone II A inhibited Tat‐induced HIV‐1 LTR transactivation dependent on AMPK‐Nampt pathway. Collectively, our data provide new insights into understanding of the molecular mechanisms of tanshinone II A inhibited Tat‐regulated transcription, suggesting that targeting AMPK/Nampt/SIRT1 pathway could serve as new anti‐HIV‐1 agents. J. Cell. Physiol. 229: 1193–1201, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24552   open full text
• Protective Effect of the Y220C Mutant p53 Against Steatosis: Good News?
  Manuele Gori, Barbara Barbaro, Mario Arciello, Roberta Maggio, Carmela Viscomi, Alessia Longo, Clara Balsano.
  Journal of Cellular Physiology. May 23, 2014
  Nonalcoholic fatty liver disease (NAFLD) ranges from simple steatosis to steatohepatitis, which may progress to fibrosis, and cirrhosis, leading eventually to hepatocarcinoma development. Recently, cases of hepatocarcinoma have been diagnosed in steatotic patients without nonalcoholic steatohepatitis (NASH) and cirrhosis. The p53 protein, besides its function as tumor suppressor, is emerging as an important regulator of cellular metabolism, but its role in steatosis remains unclear. We induced steatosis in HepG2 (wt‐p53) and Huh7.5.1 (Y220C‐mutant p53) cells using free fatty acids. We observed a different modulation of p53, different intracellular lipid content, and similar down‐regulation of the de novo lipid synthesis genes but opposite modulation of the fatty acid β‐oxidation pathway between HepG2 and Huh7.5.1. Accordingly, we found a diverse amount of apoptosis and reactive oxygen species between the two cell lines. Transfection of the wt‐p53 in Huh7.5.1 cells reverted the different lipid metabolism behavior observed in these cells. In conclusion, unlike the wt‐p53, the Y220C mutant provides a specific protection against steatosis and potentially against its progression. Our findings highlight for the first time an unknown role of a p53 mutant in the setting of steatosis. Being this mutation very frequent in human cancers, this study could be a breakthrough in explaining the occurrence of hepatocarcinoma in steatotic patients without NASH and cirrhosis. J. Cell. Physiol. 229: 1182–1192, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24550   open full text
• Definition of Novel Electrochemotherapy Parameters and Validation of their in Vitro and in Vivo Effectiveness.
  Enrico P. Spugnini, Alessandro Melillo, Lucio Quagliuolo, Mariarosaria Boccellino, Bruno Vincenzi, Paola Pasquali, Alfonso Baldi.
  Journal of Cellular Physiology. May 23, 2014
  Electrochemotherapy (ECT) is a cancer therapy that conjugates the administration of a chemotherapy agent to the delivery of permeabilizing pulses released singularly or as bursts. This approach results in higher number of anticancer molecules delivered to their biological targets, but is also associated to undesirable side effects such as pain and muscular contractions. A new electroporator delivering train of eight biphasic pulses at the voltage of 1,300 V/cm lasting 50 + 50 µsec each, with a frequency of 1 Hz, and with 10‐µsec interpulse intervals (total treatment time: 870 µsec/cm2 of treated area) was tested in vitro on the human lung cancer cell line A549 and in vivo, both in mice xenografts and privately owned rabbits with spontaneous tumors. The tumor cell line was treated with electroporation using the new parameters, that showed improved drug efficacy in causing cell death. Mice with chemoresistant xenografts were treated as well with either the new parameters and with a previous protocol, confirming the higher tolerability and efficacy of the novel parameters. Finally, a cohort of six pet rabbits with advanced skin neoplasms were enrolled in a compassionate trial using the new parameters in adjuvant fashion. In terms of efficacy, none of the rabbits experienced tumor recurrence, showing minimal discomfort during the ECT sessions. The data described, demonstrate that the new permeabilizing protocol adopting biphasic electric pulses displays a significant higher efficacy compared to previous ECT treatments and substantial reduction of the associated morbidity. J. Cell. Physiol. 229: 1177–1181, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24548   open full text
• Dependence of Castration‐Resistant Prostate Cancer (CRPC) Stem Cells on CRPC‐Associated Fibroblasts.
  Helty Adisetiyo, Mengmeng Liang, Chun‐Peng Liao, Joseph H. Jeong, Michael B. Cohen, Pradip Roy‐Burman, Baruch Frenkel.
  Journal of Cellular Physiology. May 23, 2014
  We previously established a role for cancer‐associated fibroblasts (CAF) in enhancing the self‐renewal and differentiation potentials of putative prostate cancer stem cells (CSC). Our published work focused on androgen‐dependent prostate cancer (ADPC) using the conditional Pten deletion mouse model. Employing the same model, we now describe the interaction of CAF and CSC in castration‐resistant prostate cancer (CRPC). CAF isolated from ADPC (ADPCAF) and from CRPC (CRPCAF) were compared in terms of their ability to support organoid formation and tumor initiation by CSC from CRPC (CRPCSC) in vitro and in vivo. CRPCSC formed spheroids in vitro and well‐differentiated glandular structures under the renal capsules of recipient mice in vivo more effectively in the presence of CRPCAF compared to ADPCAF. Furthermore, whereas CSC with CAF from ADPC formed mostly well‐differentiated tumors in our previous study, we now show that CRPCSC, when combined with CRPCAF (but not ADPCAF), can form aggressive, poorly‐differentiated tumors. The potential of CRPCAF to support organoid/tumor formation by CRPCSC remained greater even when compared to 10‐fold more ADPCAF, suggesting that paracrine factors produced specifically by CRPCAF preferentially potentiate the stemness and tumorigenic properties of the corresponding CSC. This apparently unique property of CRPCAF was notable when the CAF and CSC were grafted in either intact or castrated recipient mice. In both environments, CRPCAF induced in the epithelial compartment higher proliferative activity compared to ADPCAF, indicated by a higher Ki67 index. Factors released by CRPCAF to regulate CRPCSC may be targeted to develop novel therapeutic approaches to manage advanced prostate cancer. J. Cell. Physiol. 229: 1170–1176, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24546   open full text
• SKP2 Overexpression Is Associated With Increased Serine 10 Phosphorylation of p27 (pSer10p27) in Triple‐Negative Breast Cancer.
  Katerina D. Fagan‐Solis, Brian T. Pentecost, Joseph M. Gozgit, Brooke A. Bentley, Sharon M. Marconi, Christopher N. Otis, Douglas L. Anderton, Sallie Smith Schneider, Kathleen F. Arcaro.
  Journal of Cellular Physiology. May 23, 2014
  S‐phase kinase‐associated protein 2 (SKP2) is an important cell cycle regulator, targeting the cyclin‐dependent kinase (CDK) inhibitor p27 for degradation, and is frequently overexpressed in breast cancer. p27 regulates G1/S transition by abrogating the activity of cyclin/CDK complexes. p27 can undergo phosphorylation at serine 10 (pSer10p27). This phosphorylation event is associated with increased cell proliferation and poor prognosis in patients with glioma. The relationship between SKP2 and pSer10p27 in breast cancer has not been previously investigated. Immunohistochemistry (IHC) of SKP2, p27, pSer10p27, and other genes involved in this pathway, was analyzed in 188 breast tumors and 50 benign reduction mammoplasty samples. IHC showed SKP2 to be more highly expressed in estrogen receptor α (ERα)‐negative breast cancers and demonstrated that triple‐negative tumors were more likely to have high expression of SKP2 than were non‐triple negative, ERα‐negative tumors. A significant positive relationship was discovered for SKP2 and pSer10p27. High levels of SKP2 and pSer10p27 were observed significantly more often in ERα‐negative and triple‐negative than in ERα‐positive breast cancers. Use of the triple‐negative TMX2‐28 breast cancer cell line to address the role of SKP2 in cell cycle progression confirmed that SKP2 contributes to a more rapid cell cycle progression and may regulates pSer10p27 levels. Together, the results indicate that presence of high SKP2 plus high pSer10p27 levels in triple‐negative breast cancers is associated with aggressive growth, and highlight the validity of using SKP2 inhibitors as a therapeutic approach for treating this subset of breast cancers. J. Cell. Physiol. 229: 1160–1169, 2014. © 2014 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24545   open full text
• Basic Models Modeling Resistance Training: An Update for Basic Scientists Interested in Study Skeletal Muscle Hypertrophy.
  Jason Cholewa, Lucas Guimarães‐Ferreira, Tamiris da Silva Teixeira, Marshall Alan Naimo, Xia Zhi, Rafaele Bis Dal Ponte de Sá, Alice Lodetti, Mayara Quadros Cardozo, Nelo Eidy Zanchi.
  Journal of Cellular Physiology. May 23, 2014
  Human muscle hypertrophy brought about by voluntary exercise in laboratorial conditions is the most common way to study resistance exercise training, especially because of its reliability, stimulus control and easy application to resistance training exercise sessions at fitness centers. However, because of the complexity of blood factors and organs involved, invasive data is difficult to obtain in human exercise training studies due to the integration of several organs, including adipose tissue, liver, brain and skeletal muscle. In contrast, studying skeletal muscle remodeling in animal models are easier to perform as the organs can be easily obtained after euthanasia; however, not all models of resistance training in animals displays a robust capacity to hypertrophy the desired muscle. Moreover, some models of resistance training rely on voluntary effort, which complicates the results observed when animal models are employed since voluntary capacity is something theoretically impossible to measure in rodents. With this information in mind, we will review the modalities used to simulate resistance training in animals in order to present to investigators the benefits and risks of different animal models capable to provoke skeletal muscle hypertrophy. Our second objective is to help investigators analyze and select the experimental resistance training model that best promotes the research question and desired endpoints. J. Cell. Physiol. 229: 1148–1156, 2014. © 2013 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24542   open full text
• New Advances of microRNAs in Glioma Stem Cells, With Special Emphasis on Aberrant Methylation of microRNAs.
  Bing Zhao, Er‐Bao Bian, Jia Li, Jun Li.
  Journal of Cellular Physiology. May 23, 2014
  Malignant brain tumors are thought to be originate from a small population of cells that display stem cell properties, including the capacity of self‐renewal, multipotent differentiation, initiation of tumor tissues. Cancer stem cells (CSCs) have been identified in gliomas in which they are named as glioma stem cells (GSCs). GSCs, sharing some characteristics with normal neural stem cells (NSCs), contribute to the cellular origin for primary gliomas and the recurrence of malignant gliomas after current conventional therapy. Recently, increasing evidences have showed that miRNAs play a central role in GSCs. In this review we focus on the role of GSCs in gliomas and in the abnomal expression of miRNAs in GSCs. Furthermore, we also discuss epigenetic dysregulation of tumor‐suppressor miRNAs by promoter DNA methylation is involved in the regulation of GSCs biology. Recent advances in understanding dysregulated expression of miRNAs and methylation of tumor‐suppressor miRNAs in GSCs and their possible use as new therapeutic targets of gliomas. J. Cell. Physiol. 229: 1141–1147, 2014. © 2013 Wiley Periodicals, Inc.

  May 23, 2014   doi: 10.1002/jcp.24540   open full text
• Ras Oncoprotein Disrupts the TSH/CREB Signaling Upstream Adenylyl Cyclase in Human Thyroid Cell.
  Marcella Salzano, Eleonora Russo, Salvatore Salzano, Maurizio Bifulco, Mario Vitale.
  Journal of Cellular Physiology. May 13, 2014
  Activating mutations in RAS genes and p21 Ras overactivation are common occurrences in a variety of human tumors. p21 Ras oncoproteins deregulate a number of signaling pathways, dedifferentiating the thyroid cell, and negatively regulating the expression of thyroid specific genes. In rat thyroid cells, Ras oncoproteins inhibit the TSH pathway by reducing PKA activity and thus the expression of thyroid specific genes, while in mouse melanocytes, Ras oncoproteins reduce the αMSH‐stimulated cAMP signaling by increasing the expression of the phosphodiesterase‐4B. Given these cell‐dependent differences, we investigated if and how the TSH/CREB pathway is modulated by Ras oncoprotein in a human thyroid cell line. CREB phosphorylation was stimulated by TSH and forskolin in TAD‐2 cells. RasV12 expression negatively regulated the TSH‐stimulated CREB phosphorylation but was ineffective on forskolin‐stimulated CREB phosphorylation. Phosphodiesterase inhibition by IBMX enhanced TSH‐stimulated CREB phosphorylation, but did not restore TSH‐stimulated CREB phosphorylation inhibited by Ras oncoprotein. These data indicate that Ras oncoprotein disrupts the TSH/CREB pathway, upstream adenylyl cyclase, and highlight the existence of mechanisms of interaction between Ras and the cAMP pathway different in human and in rat thyroid cells. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24672   open full text
• A p53‐dependent tumor suppressor network is induced by selective miR‐125a‐5p inhibition in multiple myeloma cells in vitro.
  Marzia Leotta, Lavinia Biamonte, Lavinia Raimondi, Domenica Ronchetti, Maria Teresa Di Martino, Cirino Botta, Emanuela Leone, Maria Rita Pitari, Antonino Neri, Antonio Giordano, Pierosandro Tagliaferri, Pierfrancesco Tassone, Nicola Amodio.
  Journal of Cellular Physiology. May 13, 2014
  The analysis of deregulated microRNAs (miRNAs) is emerging as a novel approach to disclose the regulation of tumor suppressor or tumor promoting pathways in tumor cells. Targeting aberrantly expressed miRNAs is therefore a promising strategy for cancer treatment. By miRNA profiling of primary plasma cells from multiple myeloma (MM) patients, we previously reported increased miR‐125a‐5p levels associated to specific molecular subgroups. On these premises, we aimed at investigating the biological effects triggered by miR‐125a‐5p modulation in MM cells. Expression of p53 pathway‐related genes was down‐regulated in MM cells transfected with miR‐125a‐5p mimics. Luciferase reporter assays confirmed specific p53 targeting at 3'UTR level by miR‐125a‐5p mimics. Interestingly, bone marrow stromal cells (BMSCs) affected the miR‐125a‐5p/p53 axis, since adhesion of MM cells to BMSCs strongly up‐regulated miR‐125a‐5p levels, while reduced p53 expression. Moreover, ectopic miR‐125a‐5p reduced, while miR‐125–5p inhibitors promoted, the expression of tumor suppressor miR‐192 and miR‐194, transcriptionally regulated by p53. Lentiviral‐mediated stable inhibition of miR‐125a‐5p expression in wild‐type p53 MM cells dampened cell growth, increased apoptosis and reduced cell migration. Importantly, inhibition of in vitro MM cell proliferation and migration was also achieved by synthetic miR‐125a‐5p inhibitors and was potentiated by the co‐expression of miR‐192 or miR‐194. Taken together, our data indicate that miR‐125a‐5p antagonism results in the activation of p53 pathway in MM cells, underlying the crucial role of this miRNA in the biopathology of MM and providing the molecular rationale for the combinatory use of miR‐125a inhibitors and miR‐192 or miR‐194 mimics for MM treatment. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24669   open full text
• Expression of Adenine Nucleotide Translocase (ANT) Isoform Genes Is Controlled by Peroxisome Proliferator‐Activated Receptor‐γ Coactivator 1α (PGC‐1α) Through Different Transcription Factors.
  Aleix Gavaldà‐Navarro, Josep A. Villena, Anna Planavila, Octavi Viñas, Teresa Mampel.
  Journal of Cellular Physiology. May 13, 2014
  Adenine nucleotide translocase (ANT) isoforms are mitochondrial proteins encoded by nuclear DNA that catalyzes the exchange of ATP generated in the mitochondria for ADP produced in the cytosol. The aim of this study was to determine the role of the transcriptional coactivator PGC‐1α (peroxisome proliferator‐activated receptor‐γ [PPAR‐γ] coactivator 1α), a master regulator of mitochondrial oxidative metabolism, in the regulation of the expression of ANT isoform genes and to identify the transcription factors involved. We found that PGC‐1α overexpression induced the expression of all ANT human and mouse isoforms but to different degrees. The transcription factor ERRα was involved in PGC‐1α‐induced expression of all human ANT isoforms (hANT1‐3) in HeLa cells as well as in the regulation of mouse isoforms (mANT1‐2) in C2C12 myotubes and 3T3‐L1 adipocytes, even though ANT isoforms have important physiological differences and are regulated in a tissue‐specific manner. In addition to ERRα, PPARδ and mTOR pathways were involved in the induction of mANT1‐2 by PGC‐1α in C2C12 myotubes, while PPARγ was involved in PGC‐1α‐regulation of mANT1‐2 in 3T3‐L1 adipocytes. Furthermore, the regulation of mANT genes by PGC‐1α was also observed in vivo in knockout mouse models lacking PGC‐1α. In summary, our results show that the regulation of genes encoding ANT isoforms is controlled by PGC‐1α through different transcription factors depending on cell type. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24671   open full text
• High Insulin Level‐Induced Down‐Regulation of Erk‐1/IGF‐1R/FGFR‐1 Signaling Is Required for Oxidative Stress‐Mediated Apoptosis of Adipose Tissue‐Derived Stem Cells.
  Maria Giovanna Scioli, Valerio Cervelli, Gaetano Arcuri, Pietro Gentile, Elena Doldo, Alessandra Bielli, Elena Bonanno, Augusto Orlandi.
  Journal of Cellular Physiology. May 13, 2014
  Homeostasis of adipose tissue requires highly coordinated response between circulating factors and cell population. Human adult adipose tissue‐derived stem cells (ASCs) display multiple differentiation properties and are sensitive to insulin stimulation. Insulin resistance and high level of circulating insulin characterize patients with type 2 diabetes and obesity. At physiological concentration, insulin promoted proliferation and survival of ASCs in vitro, whereas high insulin level induced their dose‐dependent proliferative arrest and apoptosis. Insulin‐induced apoptotic commitment depended on the down‐regulation of Erk‐1, insulin growth factor‐1 receptor (IGF‐1R), and fibroblast growth factor receptor‐1 (FGFR‐1)‐mediated signaling. Specific inhibition of Erk‐1/2, IGF‐1R, and FGFR activity promoted ASC apoptosis but did not increase insulin effects, whereas EGFR and ErbB2 inhibition potentiated insulin‐induced apoptosis. FGFRs and EGFR inhibition reduced ASC adipogenic differentiation, whereas Erk‐1/2 and IGF‐1R inhibition was ineffective. Insulin‐induced apoptosis associated to reactive oxygen species (ROS) accumulation and inhibition of NADPH oxidase 4 (Nox4) activity prevented ASC apoptosis. Moreover, specific inhibition of Erk‐1/2, IGF‐1R, and FGFR‐1 activity promoted ROS generation and this effect was not cumulative with that of insulin alone. Our data indicate that insulin concentration is a critical regulatory switch between proliferation and survival of ASCs. High insulin level‐induced apoptotic machinery involves Nox4‐generated oxidative stress and the down‐regulation of a complex receptor signaling, partially distinct from that influencing adipogenic differentiation of ASCs. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24667   open full text
• mTORC1 Is Involved in Hypoxia‐Induced Pulmonary Hypertension through the Activation of Notch3.
  Wang Wang, Jie Liu, Aiping Ma, Ran Miao, Yuling Jin, Hongbing Zhang, Kaifeng Xu, Chen Wang, Jun Wang.
  Journal of Cellular Physiology. May 13, 2014
  Hypoxia‐induced pulmonary hypertension (HPH) is a clinical syndrome associated with high morbidity and mortality. However, the underlying mechanisms remain unclear. Both the mammalian target of rapamycin (mTOR) and the Notch3 signaling pathways have been reported to be involved in HPH; however, it is unknown whether there is a connection between these two signaling pathways in HPH. This study was designed to investigate the relationship between mTOR and Notch3 in HPH. After treatment with 10% O2 for 4 weeks, male C57BL/6 mice developed HPH with gradually increased right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), and pulmonary arteriolar remodeling accompanied by the activation of mTOR complex 1 (mTORC1) and Notch3 in the lung tissue and pulmonary arterioles. Pretreatment with the mTORC1 inhibitor rapamycin not only alleviated pulmonary arterial pressure and pulmonary arteriolar remodeling but also suppressed hypoxia‐induced mTORC1 and Notch3 activation. Prophylactic N‐[N‐(3,5‐difluorophenacetyl)‐L‐alanyl]‐S‐phenylglycine t‐butyl ester (DAPT) administration, a Notch signaling inhibitor, protected against the effects of hypoxia. These in vivo data were confirmed by in vitro experiments on human pulmonary arterial smooth muscle cell (PASMC) exposed to 3% O2. Furthermore, overexpression of Notch3 intracellular domain partially abrogated the inhibitory effects of rapamycin on human PASMC proliferation. These data indicate that both mTORC1 and Notch3 signaling are involved in HPH and the downstream effects of mTORC1 activation in HPH are partially dependent on the activation of Notch3 signaling. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24670   open full text
• Bradykinin regulates osteoblast differentiation by Akt/ERK/NFκB signalling axis.
  Swati Srivastava, Kirti Sharma, Narender Kumar, Partha Roy.
  Journal of Cellular Physiology. May 13, 2014
  Bradykinin (BK), a well known mediator of pain and inflammation, is also known to be involved in the process of bone resorption. The present study therefore evaluated the role of BK in osteoblast lineage commitment. Our data showed that BK inhibits the migration of bone marrow mesenchymal stem cells, but does not affect their viability. Moreover, BK also inhibits osteoblastic differentiation by significantly downregulating the levels of mRNAs for osteopontin, runX2, col24, osterix, osteocalcin genes and bone mineralization (p  < 0.05). Further, BK was found to elicit the BK receptors (BDKR1 and BDKR2) mediated activation of ERK1/2 and Akt pathways, which finally led to the activation of NFκB. BK also promoted the osteoclast differentiation of bone marrow derived preosteoclast cells by upregulating the expression of c‐fos, NFATC1, TRAP, clcn7, cathK, and OSCAR genes and increasing TRAP activity through NFκB pathway. In conclusion, our data suggests that BK decreases the differentiation of osteoblasts with concomitant increase in osteoclast formation and thus provides new insight into the mechanism of action of BK in modulating bone resorption. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 13, 2014   doi: 10.1002/jcp.24668   open full text
• Short‐Term Hypoxia Enhances the Migratory Capability of Dendritic Cell Through HIF‐1α and PI3K/Akt Pathway.
  Irene Filippi, Emilia Morena, Carlo Aldinucci, Fabio Carraro, Silvano Sozzani, Antonella Naldini.
  Journal of Cellular Physiology. May 12, 2014
  Hypoxia represents an inadequate oxygen supply to tissues, which can modulate cell functions, primarily through the hypoxia‐inducible transcription factor HIF‐1α. Dendritic cells (DC) are professional antigen‐presenting cells and their migration maybe affected by hypoxia, since the local microenvironment in lymphoid organs, as well as in inflamed and tumor tissues, is characterized by low oxygen tensions. In this study we observed an enhanced migratory capability of human monocyte‐derived DC, using in vitro migration assays performed under hypoxic conditions. Such enhancement was independent on either the chemoattractants involved or the maturation level of DC. However, HIF‐1α appeared to be crucial for the migration only of immature DC and not for mature DC under hypoxia, as indicated by HIF‐1α siRNA approaches. Furthermore, we observed that while Akt phosphorylation was enhanced in both immature and mature DC exposed to hypoxia, other signaling pathways, such as p38 and p42/p44 MAPK, were differently affected during hypoxic treatment. More interestingly, aspecific and specific inhibition of PI3K/Akt indicated that such pathway was relevant for the migration of both immature and matured DC under hypoxia, even when DC were transfected with HIF‐1α siRNA. Our results indicate that, besides HIF‐1α, several other pathways, including PI3K/Akt, may be involved in the response to hypoxia of immature and, more specifically, of mature DC to sustain their trafficking and functions within hypoxic microenvironments. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  May 12, 2014   doi: 10.1002/jcp.24666   open full text
• Negative Regulators of Brown Adipose Tissue (BAT)‐Mediated Thermogenesis.
  Bal Krishan Sharma, Mallikarjun Patil, Ande Satyanarayana.
  Journal of Cellular Physiology. May 08, 2014
  Brown adipose tissue (BAT) is specialized for energy expenditure, a process called adaptive thermogenesis. PET‐CT scans recently demonstrated the existence of metabolically active BAT in adult humans, which revitalized our interest in BAT. Increasing the amount and/or activity of BAT holds tremendous promise for the treatment of obesity and its associated diseases. PGC1α is the master regulator of UCP1‐mediated thermogenesis in BAT. A number of proteins have been identified to influence thermogenesis either positively or negatively through regulating the expression or transcriptional activity of PGC1α. Therefore, BAT activation can be achieved by either inducing the expression of positive regulators of PGC1α or by inhibiting the repressors of the PGC1α/UCP1 pathway. Here, we review the most important negative regulators of PGC1α/UCP1 signaling and their mechanism of action in BAT‐mediated thermogenesis. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 08, 2014   doi: 10.1002/jcp.24664   open full text
• RUNX3 Facilitates Growth of Ewing Sarcoma Cells.
  Krista L. Bledsoe, Meghan E. McGee‐Lawrence, Emily T. Camilleri, Xiaoke Wang, Andre J. van Wijnen, Andre M. Oliveira, Jennifer J. Westendorf.
  Journal of Cellular Physiology. May 08, 2014
  Ewing sarcoma is an aggressive pediatric small round cell tumor that predominantly occurs in bone. Approximately 85% of Ewing sarcomas harbor the EWS/FLI fusion protein, which arises from a chromosomal translocation, t(11:22)(q24:q12). EWS/FLI interacts with numerous lineage‐essential transcription factors to maintain mesenchymal progenitors in an undifferentiated state. We previously showed that EWS/FLI binds the osteogenic transcription factor RUNX2 and prevents osteoblast differentiation. In this study, we investigated the role of another Runt‐domain protein, RUNX3, in Ewing sarcoma. RUNX3 participates in mesenchymal‐derived bone formation and is a context dependent tumor suppressor and oncogene. RUNX3 was detected in all Ewing sarcoma cells examined, whereas RUNX2 was detected in only 73% of specimens. Like RUNX2, RUNX3 binds to EWS/FLI via its Runt domain. EWS/FLI prevented RUNX3 from activating the transcription of a RUNX‐responsive reporter, p6OSE2. Stable suppression of RUNX3 expression in the Ewing sarcoma cell line A673 delayed colony growth in anchorage independent soft agar assays and reversed expression of EWS/FLI‐responsive genes. These results demonstrate an important role for RUNX3 in Ewing sarcoma. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 08, 2014   doi: 10.1002/jcp.24663   open full text
• Bisphenol‐A Induces Podocytopathy With Proteinuria in Mice.
  Nuria Olea‐Herrero, María Isabel Arenas, Carmen Muñóz‐Moreno, Rafael Moreno‐Gómez‐Toledano, Marta González‐Santander, Ignacio Arribas, Ricardo J. Bosch.
  Journal of Cellular Physiology. May 08, 2014
  Bisphenol‐A, a chemical used in the production of the plastic lining of food and beverage containers, can be found in significant levels in human fluids. Recently, bisphenol‐A has been associated with low‐grade albuminuria in adults as well as in children. Since glomerular epithelial cells (podocytes) are commonly affected in proteinuric conditions, herein we explored the effects of bisphenol‐A on podocytes in vitro and in vivo. On cultured podocytes we first observed that bisphenol‐A—at low or high concentrations—(10 nM and 100 nM, respectively) was able to induce hypertrophy, diminish viability, and promote apoptosis. We also found an increase in the protein expression of TGF‐β1 and its receptor, the cyclin‐dependent kinase inhibitor p27Kip1, as well as collagen‐IV, while observing a diminished expression of the slit diaphragm proteins nephrin and podocin. Furthermore, mice intraperitoneally injected with bisphenol‐A (50 mg/Kg for 5 weeks) displayed an increase in urinary albumin excretion and endogenous creatinine clearance. Renal histology showed mesangial expansion. At ultrastructural level, podocytes displayed an enlargement of both cytoplasm and foot processes as well as the presence of condensed chromatin, suggesting apoptosis. Furthermore, immunohistochemistry for WT‐1 (specific podocyte marker) and the TUNEL technique showed podocytopenia as well as the presence of apoptosis, respectively. In conclusion, our data demonstrate that Bisphenol‐A exposure promotes a podocytopathy with proteinuria, glomerular hyperfiltration and podocytopenia. Further studies are needed to clarify the potential role of bisphenol‐A in the pathogenesis as well as in the progression of renal diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 08, 2014   doi: 10.1002/jcp.24665   open full text
• Big Data Bioinformatics.
  Casey S. Greene, Jie Tan, Matthew Ung, Jason H. Moore, Chao Cheng.
  Journal of Cellular Physiology. May 06, 2014
  Recent technological advances allow for high throughput profiling of biological systems in a cost‐efficient manner. The low cost of data generation is leading us to the “big data” era. The availability of big data provides unprecedented opportunities, but it also brings out challenges in data mining and analysis. In this review, we introduce key concepts in the analysis of big data, including both “machine learning” algorithms as well as “unsupervised” and “supervised” examples of each. We note packages for the R programming language that are available perform machine learning analyses. In addition to programming based solutions, we review webservers that allow users with limited or no programming background to perform these analyses on large data compendia. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  May 06, 2014   doi: 10.1002/jcp.24662   open full text
• Reactive Oxygen Species‐Dependent Nitric Oxide Production in Reciprocal Interactions of Glioma and Microglial Cells.
  Shing‐Chuan Shen, Ming‐Shun Wu, Hui‐Yi Lin, Liang‐Yo Yang, Yi‐Hsuan Chen, Yen‐Chou Chen.
  Journal of Cellular Physiology. April 29, 2014
  Conditional mediums (CMs) from glioma cells U87, GBM‐8401, and C6 significantly induced iNOS protein and NO production by microglial cells BV‐2 but without altering the cell viability or cell‐cycle progression of BV2 microglia. Significant increases in intracellular peroxide by U87‐CM and C6‐CM were detected by a DCHF‐DA assay, and vitamin (Vit) C and N‐acetyl cysteine (NAC)‐reduced intracellular peroxide levels elicited by CMs lead to inhibition of iNOS/NO production The extracellular signal‐regulated kinase (ERK) inhibitor, U0126, and c‐Jun N‐terminal kinase (JNK) inhibitor, SP600125, suppressed U87‐CM‐ and C6‐CM‐induced iNOS/NO production by respectively blocking phosphorylated ERK (pERK) and JNK (pJNK) protein expressions stimulated by U87‐CM and C6‐CM. Increased migration of U87 and C6 glioma cells by a co‐culture with BV‐2 microglial cells or adding the nitric oxide (NO) donor, sodium nitroprusside (SNP) was observed, and that was blocked by adding an NO synthase (NOS) inhibitor, N‐nitro l‐arginine methyl ester (NAME). Contributions of ROS, pERK, and pJNK to the migration of glioma cells was further demonstrated in a transwell coculture system of U87 and C6 gliomas with BV‐2 microglial cells. Furthermore, expressions of tumor necrosis factor (TNF)‐α and monocyte chemoattractant protein (MCP)‐1 messenger (m)RNA in U87 and C6 cells were detected by an RT‐PCR, and TNF‐α and MCP‐1 induced iNOS protein expression in time‐ and concentration‐dependent manners. Neutralization of TNF‐α or MCP‐1 in U87‐CM and C6‐CM using a TNF‐α or MCP‐1 antibody inhibited iNOS protein expression, and increased intracellular peroxide by TNF‐α or MCP‐1 was identified in BV‐2 cells. The reciprocal activation of glioma cells and microglia via ROS‐dependent iNOS/NO elevation at least partially mediated by TNF‐α and MCP‐1 is elucidated. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 29, 2014   doi: 10.1002/jcp.24659   open full text
• Statin‐Induced Impairment of Monocyte Migration Is Gender‐Related.
  Anna Ruggieri, Lucrezia Gambardella, Angela Maselli, Rosa Vona, Simona Anticoli, Alessia Panusa, Walter Malorni, Paola Matarrese.
  Journal of Cellular Physiology. April 29, 2014
  Statins, widely used for treatment of hypercholesterolemia, have been demonstrated to exert pleiotropic beneficial effects independently of their cholesterol‐lowering action, such as anti‐inflammatory activity. A gender disparity has been observed in their cholesterol lowering activity as well as in response to these “off label” effects. Monocytes play a central role in atherosclerotic disease and, more in general, in inflammatory responses, through their chemotactic function and cytokine production. On these bases, in the present work, we examined the effect of statins on homeostasis and migration properties of freshly isolated monocytes from male and female healthy donors. Two prototypic natural and synthetic statins with different polarity, that is, type 1 and type 2 statins, have been considered: simvastatin and atorvastatin. Freshly isolated monocytes from peripheral blood of male and female healthy donors were treated with these drugs in the absence or presence of lipopolysaccharide (LPS) stimulation. Results obtained indicated that the polar statin efficiently inhibited chemotaxis of monocytes more than the apolar statin and that this effect was more significantly induced in cells from females than in cells from males. Dissecting the mechanisms involved, we found that these results could mainly be due to differential effects on: (i) the release of key cytokines, for example, MCP‐1 and TNF‐α; (ii) the maintenance of the redox homeostasis; (iii) a target activity on microfilament network integrity and function. All in all these results could suggest a reappraisal of “off‐label” effects of statins taking into account either their chemical structure, that is, molecular polarity, or the gender issue. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 29, 2014   doi: 10.1002/jcp.24657   open full text
• Expression Variability and Function of the RET Gene in Adult Peripheral Blood Mononuclear Cells.
  Marta Rusmini, Paola Griseri, Ivana Matera, Elena Pontarini, Roberto Ravazzolo, Domenico Mavilio, Isabella Ceccherini.
  Journal of Cellular Physiology. April 29, 2014
  RET is a gene playing a key role during embryogenesis and in particular during the enteric nervous system development. High levels of RET gene expression are maintained in different human tissues also in adulthood, although their physiological role remains unclear. In particular, collected evidences of an RET contribution in the development and maintenance of the immune system prompted us to investigate its levels of surface expression on peripheral blood mononuclear cells (PBMCs) from adult healthy donors. Despite variability among samples, RET expression was conserved at similar levels in the different immune cell subsets, with higher correlations in similar lymphocyte populations (i.e. CD4+ and CD8+ T cells). Conversely, no correlation was found between the amount of RET receptor, the expression of its putative ligands and co‐receptors and the genotypes at the RET locus. Moreover, we investigated the RET‐associated inflammatory pathways in PBMCs from healthy donors both in resting conditions and upon glial cell derived neurotrophic factor (GDNF) and GPI‐linked co‐receptors alpha 1 (GFRα1) mediated RET activation. RET mRNA levels positively correlated with the transcript amount of interleukin‐8 (IL‐8), a cytokine produced by monocytes and macrophages, though we could not demonstrate its direct effect on RET expression by in vitro experiments on THP1 human monocytic cells. These results imply that RET expression might be influenced by either cis‐ and/or trans‐factors, which together would account for its high variability within the general population, and suggest a putative functional role of the RET gene in modulating immune cell responses during inflammation and carcinogenesis. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 29, 2014   doi: 10.1002/jcp.24660   open full text
• Sub‐Toxic Nicotine Concentrations Affect Extracellular Matrix and Growth Factor Signaling Gene Expressions in Human Osteoblasts.
  Lorella Marinucci, Maria Bodo, Stefania Balloni, Paola Locci, Tiziano Baroni.
  Journal of Cellular Physiology. April 29, 2014
  Exposure to nicotine and other compounds contained in cigarette smoking affects human health. This study examined the effects of exposure to a single or multiple sub‐toxic nicotine concentrations on human osteoblasts. Cell growth and expression of genes involved in bone differentiation, extracellular matrix (ECM) metabolism, and growth factor signaling pathways were investigated in nicotine‐treated cells compared to untreated cells. Depending on osteoblast concentration and maturation stages, nicotine differently regulated cell growth. Real‐time PCR showed regulated expressions of genes expressed by nicotine‐treated osteoblasts compared to untreated cells. Among ECM genes, type I collagen was down‐regulated and osteonectin was up‐regulated in nicotine‐treated osteoblasts; similarly, fibroblast growth factor‐1 (FGF1) and fibroblast growth factor‐2 (FGF2), two members of FGF signaling system, were discordantly modulated; genes involved in osteoblast maturation and differentiation such as alkaline phosphatase (ALP), runt‐related transcription factor‐2 (RUNX2), and bone sialoprotein (BSP) were over‐expressed after drug treatment. Our results show a positive association between nicotine exposure and osteoblast phenotype and illustrate for the first time a mechanism whereby acute or chronic exposure to sub‐toxic nicotine concentrations may affect bone formation through the impairment of growth factor signaling system and ECM metabolism. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 29, 2014   doi: 10.1002/jcp.24661   open full text
• The Paracrine Feedback Loop Between Vitamin D3 (1,25(OH)2D3) and PTHrP in Prehypertrophic Chondrocytes.
  Frances C. Bach, Kirsten R.T. Rutten, Kristyanne Hendriks, Frank M. Riemers, Peter Cornelissen, Alain de Bruin, Ger J.A. Arkesteijn, Richard W. Wubbolts, William A. Horton, Louis C. Penning, Marianna A. Tryfonidou.
  Journal of Cellular Physiology. April 29, 2014
  The endocrine feedback loop between vitamin D3 (1,25(OH)2D3) and parathyroid hormone (PTH) plays a central role in skeletal development. PTH‐related protein (PTHrP) shares homology and its receptor (PTHR1) with PTH. The aim of this study was to investigate whether there is a functional paracrine feedback loop between 1,25(OH)2D3 and PTHrP in the growth plate, in parallel with the endocrine feedback loop between 1,25(OH)2D3 and PTH. This was investigated in ATDC5 cells treated with 10−8 M 1,25(OH)2D3 or PTHrP, Col2‐pd2EGFP transgenic mice, and primary Col2‐pd2EGFP growth plate chondrocytes isolated by FACS, using RT‐qPCR, Western blot, PTHrP ELISA, chromatin immunoprecipitation (ChIP) assay, silencing of the 1,25(OH)2D3 receptor (VDR), immunofluorescent staining, immunohistochemistry, and histomorphometric analysis of the growth plate. The ChIP assay confirmed functional binding of the VDR to the PTHrP promoter, but not to the PTHR1 promoter. Treatment with 1,25(OH)2D3 decreased PTHrP protein production, an effect which was prevented by silencing of the VDR. Treatment with PTHrP significantly induced VDR production, but did not affect 1α‐ and 24‐hydroxylase expression. Hypertrophic differentiation was inhibited by PTHrP and 1,25(OH)2D3 treatment. Taken together, these findings indicate that there is a functional paracrine feedback loop between 1,25(OH)2D3 and PTHrP in the growth plate. 1,25(OH)2D3 decreases PTHrP production, while PTHrP increases chondrocyte sensitivity to 1,25(OH)2D3 by increasing VDR production. In light of the role of 1,25(OH)2D3 and PTHrP in modulating chondrocyte differentiation, 1,25(OH)2D3 in addition to PTHrP could potentially be used to prevent undesirable hypertrophic chondrocyte differentiation during cartilage repair or regeneration. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 29, 2014   doi: 10.1002/jcp.24658   open full text
• Tumor Vasculature Is Regulated by FGF/FGFR Signaling‐Mediated Angiogenesis and Bone Marrow‐Derived Cell Recruitment: This Mechanism Is Inhibited by SSR128129E, the First Allosteric Antagonist of FGFRs.
  Pierre Fons, Geneviève Gueguen‐Dorbes, Jean‐Pascal Herault, Fabien Geronimi, Joël Tuyaret, Dol Frédérique, Paul Schaeffer, Cécile Volle‐Challier, Jean‐Marc Herbert, Françoise Bono.
  Journal of Cellular Physiology. April 24, 2014
  Tumor angiogenesis is also accompanied by vasculogenesis which is involved in the differentiation and mobilization of human bone marrow cells. In order to further characterize the role of vasculogenesis in the tumor growth process, the effects of FGF2 on the differentiation of human bone marrow AC133+ cells (BM‐AC133+) into vascular precursors were studied in vitro. FGF2, like VEGFA, induced progenitor cell differentiation into cell types with endothelial cell characteristics. SSR128129E, a newly discovered specific FGFR antagonist acting by allosteric interaction with FGFR, abrogated FGF2‐induced endothelial cell differentiation, showing that FGFR signaling is essential during this process. To assess the involvement of the FGF/FRGR signaling in vivo, the pre‐clinical model of Lewis lung carcinoma (LL2) in mice was used. Subcutaneous injection of LL2 cells into mice induced an increase of circulating EPCs from peripheral blood associated with tumor growth and an increase of intra‐tumoral vascular index. Treatment with the FGFR antagonist SSR128129E strongly decreased LL2 tumor growth as well as the intra‐tumoral vascular index (41% and 50% decrease vs. vehicle‐treated mice respectively, P < 0.01). Interestingly, SSR128129E treatment significantly decreased the number of circulating EPCs from the peripheral blood (53% inhibition vs. vehicle‐treated mice, P < 0.01). These results demonstrate for the first time that the blockade of the FGF/FGFR pathway by SSR128129E reduces EPC recruitment during angiogenesis‐dependent tumor growth. In this context, circulating EPCs could be a reliable surrogate marker for tumor growth and angiogenic activity. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 24, 2014   doi: 10.1002/jcp.24656   open full text
• S100A1 and S100B Expression Patterns Identify Differentiation Status of Human Articular Chondrocytes.
  Jose Diaz‐Romero, Aurelie Quintin, Eric Schoenholzer, Chantal Pauli, Alain Despont, Matthias A. Zumstein, Sandro Kohl, Dobrila Nesic.
  Journal of Cellular Physiology. April 23, 2014
  Many studies in the field of cell‐based cartilage repair have focused on identifying markers associated with the differentiation status of human articular chondrocytes (HAC) that could predict their chondrogenic potency. A previous study from our group showed a correlation between the expression of S100 protein in HAC and their chondrogenic potential. The aims of the current study were to clarify which S100 proteins are associated with HAC differentiation status and to provide an S100‐based assay for measuring HAC chondrogenic potential. The expression patterns of S100A1 and S100B were investigated in cartilage and in HAC cultured under conditions promoting dedifferentiation (monolayer culture) or redifferentiation (pellet culture or BMP4 treatment in monolayer culture), using characterized antibodies specifically recognizing S100A1 and S100B, by immunohistochemistry, immunocytochemistry, Western blot, and gene expression analysis. S100A1 and S100B were expressed homogeneously in all cartilage zones, and decreased during dedifferentiation. S100A1, but not S100B, was re‐expressed in pellets and co‐localized with collagen II. Gene expression analysis revealed concomitant modulation of S100A1, S100B, collagen type II, and aggrecan: down‐regulation during monolayer culture and up‐regulation upon BMP4 treatment. These results strongly support an association of S100A1, and to a lesser extent S100B, with the HAC differentiated phenotype. To facilitate their potential application, we established an S100A1/B‐based flow cytometry assay for accurate assessment of HAC differentiation status. We propose S100A1 and S100B expression as a marker to develop potency assays for cartilage regeneration cell therapies, and as a redifferentiation readout in monolayer cultures aiming to investigate stimuli for chondrogenic induction. J. Cell. Physiol. 229: 1106–1117, 2014. © 2014 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24547   open full text
• Hypoxia Selectively Disrupts Brain Microvascular Endothelial Tight Junction Complexes Through a Hypoxia‐Inducible Factor‐1 (HIF‐1) Dependent Mechanism.
  Sabrina Engelhardt, Abraham J. Al‐Ahmad, Max Gassmann, Omolara O. Ogunshola.
  Journal of Cellular Physiology. April 23, 2014
  The blood–brain barrier (BBB) constitutes a critical barrier for the maintenance of central nervous system homeostasis. Brain microvascular endothelial cells line the vessel walls and express tight junction (TJ) complexes that restrict paracellular passage across the BBB, thereby fulfilling a crucial role in ensuring brain function. Hypoxia, an impaired O2 delivery, is known to cause BBB dysfunction but the mechanisms that drive this disruption remain unclear. This study discloses the relevance of the master regulator of the hypoxic response, hypoxia‐inducible factor‐1 (HIF‐1), in hypoxia‐induced barrier disruption using the rat brain endothelial cell line RBE4. Hypoxic exposure rapidly induced stabilization of the HIF‐1 oxygen‐dependent alpha subunit (HIF‐1α) concomitantly with BBB impairment and TJ disruption mainly through delocalization and increased tyrosine phosphorylation of TJ proteins. Similar observations were obtained by normoxic stabilization of HIF‐1α using CoCl2, deferoxamine, and dimethyloxalylglycine underlining the involvement of HIF‐1 in barrier dysfunction particularly via TJ alterations. In agreement inhibition of HIF‐1 stabilization by 2‐methoxyestradiol and YC‐1 improved barrier function in hypoxic cells. Overall our data suggests that activation of HIF‐1‐mediated signaling disrupts TJ resulting in increased BBB permeability. J. Cell. Physiol. 229: 1096–1105, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24544   open full text
• Epithelial Immune Response in Drosophila Malpighian Tubules: Interplay Between Diap2 and Ion Channels.
  Puja Verma, Madhu G. Tapadia.
  Journal of Cellular Physiology. April 23, 2014
  Systemic immune response via the Immune deficiency pathway requires Drosophila inhibitor of apoptosis protein 2 to activate the NF‐κB transcription factor Relish. Malpighian tubules (MTs), simple epithelial tissue, are the primary excretory organs, performing additional role in providing protection to Drosophila against pathogenic infections. MTs hold a strategic position in Drosophila as one of the larval tissues that are carried over to adults, unlike other larval tissues that are histolysed during pupation. In this paper we show that Diap2 is an important regulator of local epithelial immune response in MTs and depletion of Diap2 from MTs, increases susceptibility of flies to infection. In the absence of Diap2, activation and translocation of Relish to the nucleus is abolished and as a consequence the production of IMD pathway dependent AMPs are reduced. Ion channels, (Na+/K+)‐ATPase and V‐ATPase, are important for the immune response of MTs and expression of AMPs and the IMD pathway genes are impaired on inhibition of transporters, and they restrict the translocation of Relish into the nucleus. We show that Diap2 could be regulating ion channels, as loss of Diap2 consequently reduces the expression of ion channels and affects the balance of ion concentrations which results in reduced uric acid deposition. Thus Diap2 seems to be a key regulator of epithelial immune response in MTs, perhaps by modulating ion channels. J. Cell. Physiol. 229: 1078–1095, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24541   open full text
• Phosphorylation of PRAS40‐Thr246 Involved in Renal Lipid Accumulation of Diabetes.
  Jun Hao, Fan Li, Wei Liu, Qingjuan Liu, Shuxia Liu, Hongbo Li, Huijun Duan.
  Journal of Cellular Physiology. April 23, 2014
  Lipid accumulation of kidney is a threat to renal physiological function of diabetes. The previous studies on diabetic nephropathy have demonstrated that activated Akt was involved in renal lipogenesis through enhancing transcription factor SREBP‐1. PRAS40 is one of the downstream targets of activated Akt that was reported to involve in lipid metabolism in hepatic cells. However, it is still not clear whether PRAS40 is also involved in the renal lipogenesis of diabetes. Our study revealed that phosphorylation of PRAS40‐Thr246 known as inactivated style increased in renal tubular cells of diabetic rats accompanied with over‐expression of phospho‐Akt, SREBP‐1, and ADRP. In addition, in vitro experiment also found that high glucose enhanced expression of phospho‐PRAS40‐Thr246 followed by increased SREBP‐1 and lipid droplets in HKC cells. After treated with LY294002, high glucose‐induced HKC cells showed decreased phospho‐PRAS40‐Thr246, phospho‐Akt‐Ser473, and SREBP‐1. Furthermore, wild type PRAS40 vector‐caused increased phospho‐PRAS40‐Thr246 exaggerated lipid deposits in high glucose‐treated HKC cells, which was effectively prevented in cells transfected with mutant PRAS40 vector (T246A). These above data suggested that phosphorylation of PRAS40‐Thr246 mediated abnormal lipid metabolism in kidney of diabetes and might be the potential target for treating lipogenesis of diabetic nephropathy. J. Cell. Physiol. 229: 1069–1077, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24533   open full text
• Endocrine Disruptors Differently Influence Estrogen Receptor β and Androgen Receptor in Male and Female Rat VSMC.
  Marco Pellegrini, Pamela Bulzomi, Marco Lecis, Stefano Leone, Ilaria Campesi, Flavia Franconi, Maria Marino.
  Journal of Cellular Physiology. April 23, 2014
  Sex steroid hormones differently control the major physiological processes in male and female organisms. In particular, their effects on vascular smooth muscle cells (VSMCs) migration are at the root of sex/gender‐related differences reported in the cardiovascular system. Several exogenous substances, defined endocrine disruptor chemicals (EDCs), could interfere with these androgen and estrogen effects; however, the sex/gender‐related susceptibility of VSMC motility to EDCs is completely unknown. Here, the effect of naturally occurring (naringenin, Nar) and synthetic (bisphenol A, BPA) EDCs on male and female VSMC motility has been evaluated. 17β‐estradiol (E2, 0.1 nM–1 µM) induced a dose‐dependent inhibition of motility in female‐derived VSMC. In contrast, neither dihydrotestosterone (DHT, 0.01–100 nM) nor the common precursor of sex steroid hormones, testosterone (Tes, 0.01–100 nM) modified male‐derived VSMC motility. Estrogen receptor (ER) β subtype‐dependent activation of p38 was necessary for the E2 effect on cell motility. High BPA concentration prevented E2 effects in female‐derived cells being without any effect in male‐derived cells. Nar mimicked E2 effects on female‐derived cells even in the presence of E2 or BPA. Intriguingly, Nar also inhibited the male‐derived VSMC mobility. This latter effect was prevented by ERβ inhibitor, but not by the androgen receptor (AR) inhibitor. As a whole, ERβ‐dependent signals in VSMC results more susceptible to the impact of EDCs than AR signals suggesting a possible high and overall susceptibility of female to EDCs. However, several male‐derived cells, including VSMC, express ERβ, which could also serve as target of EDC disruption in male organisms. J. Cell. Physiol. 229: 1061–1068, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24530   open full text
• Identification of the PKR Nuclear Interactome Reveals Roles in Ribosome Biogenesis, mRNA Processing and Cell Division.
  William L. Blalock, Manuela Piazzi, Alberto Bavelloni, Mirco Raffini, Irene Faenza, Antonietta D'Angelo, Lucio Cocco.
  Journal of Cellular Physiology. April 23, 2014
  The double‐strand RNA‐dependent protein kinase, PKR, plays a central role in inflammatory/chronic stress‐mediated pathologies such as cancer, diabetes, and neuro/muscular degenerative diseases. Although a significant amount of research has been conducted to elucidate the role of PKR signaling in the cytosol, only recently has attention been paid to the role of PKR in the nuclear compartment. Previously our group reported that phosphorylated forms of PKR are present in the nucleus of acute leukemic cell lines, representing a reservoir of active kinase that responds to stress. Using the CCRF‐CEM acute T‐cell leukemia cell line, a PKR‐specific inhibitor, co‐immunoprecipitation and a proteomics approach, which included affinity purified mass spectrometry analysis (AP/MS), we identified the proteins present in active and inactive PKR nuclear complexes. Of the proteins identified in the PKR complexes, sixty‐nine (69) were specific to the active complex, while thirty‐eight (38) were specific to the inactive complex. An additional thirteen (13) proteins associated specifically with both complexes. The majority of the proteins identified are involved in, ribosome biogenesis, RNA splicing, mRNA stability, gene expression, cell cycle, or chromatin organization, including several with known significance to normal hematopoiesis and/or hematological disease. In agreement with the AP/MS data, basal‐ or over‐expression of PKR under normal growth conditions favored cell proliferation in the tested cell lines, whereas pharmacological inhibition of PKR or shRNA‐mediated knock‐down did not. PKR was also found to influence the isoform and the level of expression of the proto‐oncogene MYC. J. Cell. Physiol. 229: 1047–1060, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24529   open full text
• Involvement of IRS‐1 Interaction With ADAM10 in the Regulation of Neurite Extension.
  Jin Ying Wang, Armine Darbinyan, Martyn K. White, Nune Darbinian, Krzysztof Reiss, Shohreh Amini.
  Journal of Cellular Physiology. April 23, 2014
  The insulin‐like growth factor‐1 (IGF‐1) signaling pathway plays an important role in neuronal cell differentiation. Recent studies have shown that IGF‐1 has the capacity to counteract the retraction of neuronal processes in response to inflammatory cytokines such as TNF‐α, which is a known factor for neuronal injury in the central nervous system. This event is thought to be mediated via interference of TNF‐α‐induced interaction of β1‐integrin with insulin receptor substrate‐1 (IRS‐1). Here, we demonstrate the interaction of IRS‐1 with disintegrin and metalloproteinase ADAM10 through the N‐terminal domain of IRS‐1 and that this is involved in the regulation of neurite extension and retraction by IGF‐1 and TNF‐α, respectively. PC12 cells expressing the N‐terminal domain show enhanced neurite extension after IGF‐1 treatment and reduced neurite depletion relative to control cells after TNF‐α treatment. The level of ADAM10 was found to be increased in immunohistochemical studies of HIV encephalitis clinical samples and is present with TNF‐α and TNFR1 in both astrocytes and neurons. Altogether, these observations suggest a role for ADAM10 in the mechanism for IGF1/IRS‐1 signaling pathway in sustaining the stability of neuronal processes. J. Cell. Physiol. 229: 1039–1046, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24528   open full text
• Fenretinide Induces Ubiquitin‐Dependent Proteasomal Degradation of Stearoyl‐CoA Desaturase in Human Retinal Pigment Epithelial Cells.
  William Samuel, R. Krishnan Kutty, Todd Duncan, Camasamudram Vijayasarathy, Bryan C. Kuo, Krysten M. Chapa, T. Michael Redmond.
  Journal of Cellular Physiology. April 23, 2014
  Stearoyl‐CoA desaturase (SCD, SCD1), an endoplasmic reticulum (ER) resident protein and a rate‐limiting enzyme in monounsaturated fatty acid biosynthesis, regulates cellular functions by controlling the ratio of saturated to monounsaturated fatty acids. Increase in SCD expression is strongly implicated in the proliferation and survival of cancer cells, whereas its decrease is known to impair proliferation, induce apoptosis, and restore insulin sensitivity. We examined whether fenretinide, (N‐(4‐hydroxyphenyl)retinamide, 4HPR), which induces apoptosis in cancer cells and recently shown to improve insulin sensitivity, can modulate the expression of SCD. We observed that fenretinide decreased SCD protein and enzymatic activity in the ARPE‐19 human retinal pigment epithelial cell line. Increased expression of BiP/GRP78, ATF4, and GADD153 implicated ER stress. Tunicamycin and thapsigargin, compounds known to induce ER stress, also decreased the SCD protein. This decrease was completely blocked by the proteasome inhibitor MG132. In addition, PYR41, an inhibitor of ubiquitin activating enzyme E1, blocked the fenretinide‐mediated decrease in SCD. Immunoprecipitation analysis using anti‐ubiquitin and anti‐SCD antibodies and the blocking of SCD loss by PYR41 inhibition of ubiquitination further corroborate that fenretinide mediates the degradation of SCD in human RPE cells via the ubiquitin–proteasome dependent pathway. Therefore, the effect of fenretinide on SCD should be considered in its potential therapeutic role against cancer, type‐2 diabetes, and retinal diseases. J. Cell. Physiol. 229: 1028–1038, 2014. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

  April 23, 2014   doi: 10.1002/jcp.24527   open full text
• Choline Inadequacy Impairs Trophoblast Function and Vascularization in Cultured Human Placental Trophoblasts.
  Xinyin Jiang, Sara Jones, Benjamin Y. Andrew, Anita Ganti, Olga V. Malysheva, Natasa Giallourou, Patsy M. Brannon, Mark S. Roberson, Marie A. Caudill.
  Journal of Cellular Physiology. April 23, 2014
  Maternal choline intake during gestation may influence placental function and fetal health outcomes. Specifically, we previously showed that supplemental choline reduced placental and maternal circulating concentrations of the anti‐angiogenic factor, fms‐like tyrosine kinase‐1 (sFLT1), in pregnant women as well as sFLT1 production in cultured human trophoblasts. The current study aimed to quantify the effect of choline on a wider array of biomarkers related to trophoblast function and to elucidate possible mechanisms. Immortalized HTR‐8/SVneo trophoblasts were cultured in different choline concentrations (8, 13, and 28 µM [control]) for 96‐h and markers of angiogenesis, inflammation, apoptosis, and blood vessel formation were examined. Choline insufficiency altered the angiogenic profile, impaired in vitro angiogenesis, increased inflammation, induced apoptosis, increased oxidative stress, and yielded greater levels of protein kinase C (PKC) isoforms δ and ϵ possibly through increases in the PKC activators 1‐stearoyl‐2‐arachidonoyl‐sn‐glycerol and 1‐stearoyl‐2‐docosahexaenoyl‐sn‐glycerol. Notably, the addition of a PKC inhibitor normalized angiogenesis and apoptosis, and partially rescued the aberrant gene expression profile. Together these results suggest that choline inadequacy may contribute to placental dysfunction and the development of disorders related to placental insufficiency by activating PKC. J. Cell. Physiol. 229: 1016–1027, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24526   open full text
• Platelets in Tumor Progression: A Host Factor That Offers Multiple Potential Targets in the Treatment of Cancer.
  Deva Sharma, Kathleen E. Brummel‐Ziedins, Beth A. Bouchard, Chris E. Holmes.
  Journal of Cellular Physiology. April 23, 2014
  While platelets are well known to play a central role in hemostasis and thrombosis, there is emerging experimental evidence to suggest that they also mediate tumor cell growth, dissemination, and angiogenesis. An increase in platelet number (thrombocytosis) and activity is seen in patients with a wide spectrum of malignancies, and the former is correlated with a decrease in overall survival and poorer prognosis. Preclinical data suggest that circulating tumor cell partnerships with platelets in the blood facilitate tumor metastases through direct interactions and secreted bioactive proteins. Platelets form aggregates with tumor cells, thereby protecting them from host immune surveillance through physical shielding and induction of “platelet mimicry.” There is also laboratory evidence to suggest that activated platelets interact with cancer cells within the tumor microenvironment through paracrine signaling and direct contact, thereby promoting tumor cell growth and survival. For example, platelets release mediators of both tumor angiogenesis and osteoclast resorption. The interplay between platelets and tumor cells is complex and bidirectional with involvement of multiple other components within the tumor microenvironment, including immune cells, endothelial cells, and the extracellular matrix. We review the role of platelets in tumor progression, emphasizing the opportunity these interactions afford to target platelets and platelet function to improve patient outcomes in the cancer prevention and treatment setting. J. Cell. Physiol. 229: 1005–1015, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24539   open full text
• Prohibitins Role in Cellular Survival Through Ras‐Raf‐MEK‐ERK Pathway.
  Indrajit Chowdhury, Winston E. Thompson, Kelwyn Thomas.
  Journal of Cellular Physiology. April 23, 2014
  Prohibitins are members of a highly conserved protein family containing the stomatin/prohibitin/flotillin/HflK/C (SPFH) domain (also known as the prohibitin [PHB] domain) found in unicellular eukaryotes, fungi, plants, animals, and humans. Two highly homologous members of prohibitins expressed in eukaryotes are prohibitin (PHB; B‐cell receptor associated protein‐32, BAP‐32) and prohibitin 2/repressor of estrogen receptor activity (PHB2, REA, BAP‐37). Both PHB and REA/PHB2 are ubiquitously expressed and are present in multiple cellular compartments including the mitochondria, nucleus, and the plasma membrane. Multiple functions have been attributed to the mitochondrial and nuclear PHB and PHB2/REA including cellular differentiation, anti‐proliferation, and morphogenesis. One of the major functions of the prohibitins are in maintaining the functional integrity of the mitochondria and protecting cells from various stresses. In the present review, we focus on the recent research developments indicating that PHB and PHB2/REA are involved in maintaining cellular survival through the Ras‐Raf‐MEK‐Erk pathway. Understanding the molecular mechanisms by which the intracellular signaling pathways utilize prohibitins in governing cellular survival is likely to result in development of therapeutic strategies to overcome various human pathological disorders such as diabetes, obesity, neurological diseases, inflammatory bowel disease, and cancer. J. Cell. Physiol. 229: 998–1004, 2014. © 2013 Wiley Periodicals, Inc.

  April 23, 2014   doi: 10.1002/jcp.24531   open full text
• Isoforms of Receptors of Fibroblast Growth Factors.
  Siew‐Ging Gong.
  Journal of Cellular Physiology. April 15, 2014
  The breadth and scope of Fibroblast Growth Factor signaling is immense, with documentation of its role in almost every organism and system studied so far. FGF ligands signal through a family of four distinct tyrosine kinase receptors, the FGF receptors (FGFRs). One contribution to the diversity of function and signaling of FGFs and their receptors arises from the numerous alternative splicing variants that have been documented in the FGFR literature. The present review discusses the types and roles of alternatively spliced variants of the FGFR family members and the significant impact of alternative splicing on the physiological functions of five broad classes of FGFR isoforms. Some characterized known regulatory mechanisms of alternative splicing and future directions in studies of FGFR alternative splicing are also discussed. Presence, absence, and/or the combination of specific exons within each FGFR protein impart upon each individual isoform its unique function and expression pattern during normal function and in diseased states (e.g., in cancers and birth defects). A better understanding of the diversity of FGF signaling in different developmental contexts and diseased states can be achieved through increased knowledge of the presence of specific FGFR isoforms and their impact on downstream signaling and functions. Modern high‐throughput techniques afford an opportunity to explore the distribution and function of isoforms of FGFR during development and in diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 15, 2014   doi: 10.1002/jcp.24649   open full text
• Beclin‐1 Is Required for RANKL‐Induced Osteoclast Differentiation.
  Yeon‐Ho Chung, Youngsaeng Jang, Bongkun Choi, Da‐Hyun Song, Eun‐Jin Lee, Sang‐Min Kim, Youngsup Song, Sang‐Wook Kang, Seung‐Yong Yoon, Eun‐Ju Chang.
  Journal of Cellular Physiology. April 15, 2014
  Beclin‐1 plays a critical role in autophagy; however, it also contributes to other biological processes in a non‐autophagic manner. Although studies have examined the non‐autophagic role of autophagy proteins in the secretory function of osteoclasts (OC), the role of Beclin‐1 is unclear. Here, we examined the role of Beclin‐1 in OC differentiation, and found that mouse bone marrow macrophages (BMMs) showed increased expression of Beclin‐1 upon RANKL stimulation in a p38‐ and NF‐kappa B‐dependent manner. During OC differentiation, Beclin‐1 localized to the mitochondria, where it was involved in the production of mitochondrial intracellular reactive oxygen species. Knockdown of Beclin‐1 in RANKL‐primed BMMs led to a significant reduction in RANKL‐dependent osteoclastogenesis, which was accompanied by reduced NFATc1 induction. Furthermore, knockdown of Beclin‐1 inhibited RANKL‐mediated activation of JNK and p38, both of which act downstream of reactive oxygen species, resulting in the suppression of NFATc1 induction. Finally, overexpression of constitutively active NFATc1 rescued the phenotype induced by Beclin‐1 knockdown, indicating that Beclin‐1 mediates RANKL‐induced osteoclastogenesis by regulating NFATc1 expression. These findings show that Beclin‐1 plays a non‐autophagic role in RANKL‐induced osteoclastogenesis by inducing the production of reactive oxygen species and NFATc1. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 15, 2014   doi: 10.1002/jcp.24646   open full text
• Advanced Glycation End Products Induce Lipogenesis: Regulation by Natural Xanthone through Inhibition of ERK and NF‐κB.
  Sidhartha K. Mahali, Neeharika Verma, Sunil K. Manna.
  Journal of Cellular Physiology. April 15, 2014
  Advanced glycation end products (AGE) accumulate in diabetic patients and aged persons due to high amounts of 3‐ or 4‐carbon derivatives of glucose. Understanding the mechanism of AGE‐mediated signaling leading to these consequences, like oxidative stress, inflammation, apoptosis, etc. and its regulation would be a viable strategy to control diabetic complication and age‐related diseases. We have detected the probable mechanism by which AGE increases lipogenesis, the cause of fatty liver in diabetic patients. AGE increased lipid accumulation in different cells as shown by Oil Red O staining. AGE‐mediated regulation of several transcription factors was determined by gel shift assay. Antioxidants like NAC, PDTC, and vitamin C, except mangiferin, were unable to protect AGE‐induced activation of SREBP and subsequent lipid accumulation. AGE increased the phosphorylation of ERK, and IKK and also DNA binding ability of SREBP, thereby its dependent gene transcription. AGE induces NF‐κB which might suppress PPARγ activity, in turn reducing lipid breakdown and mobilization. Mangiferin not only inhibits AGE‐mediated ROI generation that requires NF‐κB activation, but also inhibits ERK and IKK activity, thereby suppression of SREBP activity and lipogenesis. Mangiferin has shown a double‐edged sword effect to suppress AGE‐mediated ailments by reducing ROI‐mediated responses as antioxidant and inhibiting SREBP activation thereby lipogenesis, suggesting its potential efficacy against diabetes and obesity‐related diseases. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 15, 2014   doi: 10.1002/jcp.24647   open full text
• Mg2+‐dependent Modulation of BKCa Channels by Genistein in Rat Arteriolar Smooth Muscle Cells.
  Xiaoran Wang, Tingting Zhao, Shanshan Zhou, Lina Sun, Liming Zhang, Guichun Yu.
  Journal of Cellular Physiology. April 14, 2014
  Genistein, a protein tyrosine kinase (PTK) inhibitor, regulates ion channel activities. However, the mechanism of action of genistein on large‐conductance calcium‐activated potassium (BKCa) channels is unclear. This study aimed to investigate whether the mechanism of Mg2+‐dependent modulation of BKCa channel activity in vascular smooth muscle cells involved inhibition of phosphorylation by genistein or direct interaction between genistein and BKCa channels. The whole‐cell and inside‐out patch‐clamp techniques were used to measure BKCa currents and the effects of genistein on BKCa channel activities in rat mesenteric arteriolar smooth muscle cells. We found that the effects of genistein on BKCa currents were Mg2+‐dependent. Genistein (50 μM) inhibited BKCa currents if the intracellular free magnesium concentration ([Mg2+]i) was 2 μM or 20 μM, but amplified BKCa currents if [Mg2+]i was 200 μM or 2000 μM. The inhibitory effect of genistein on BKCa currents was reversed by the protein tyrosine phosphatase inhibitor sodium orthovanadate (0.5 mM). Daidzein (50 μM), an inactive analogue of genistein, also amplified BKCa currents, and its amplification was insensitive to orthovanadate. Another PTK inhibitor, tyrphostin 23 (50 μM), reduced the open probability of BKCa channels. This inhibitory effect was weaker at 200 μM [Mg2+]i than at 2 μM [Mg2+]i, and was countered by orthovanadate. Our results suggest that genistein amplifies BKCa currents at a high [Mg2+]i, but inhibits BKCa currents at a low [Mg2+]i. The mechanism of this biphasic effects involves PTK‐independent amplification and [Mg2+]i‐PTK‐dependent inhibition. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 14, 2014   doi: 10.1002/jcp.24648   open full text
• Analysis of Global Changes in Gene Expression Induced by Human Polynucleotide Phosphorylase (hPNPaseold‐35 ).
  Upneet K. Sokhi, Manny D. Bacolod, Luni Emdad, Swadesh K. Das, Catherine I. Dumur, Michael F. Miles, Devanand Sarkar, Paul B. Fisher.
  Journal of Cellular Physiology. April 14, 2014
  As a strategy to identify gene expression changes affected by human polynucleotide phosphorylase (hPNPaseold‐35 ), we performed gene expression analysis of HeLa cells in which hPNPaseold‐35 was overexpressed. The observed changes were then compared to those of HO‐1 melanoma cells in which hPNPaseold‐35 was stably knocked down. Through this analysis, 90 transcripts, which positively or negatively correlated with hPNPaseold‐35 expression, were identified. The majority of these genes were associated with cell communication, cell cycle, and chromosomal organization gene ontology categories. For a number of these genes, the positive or negative correlations with hPNPaseold‐35 expression were consistent with transcriptional data extracted from the TCGA (The Cancer Genome Atlas) expression datasets for colon adenocarcinoma (COAD), skin cutaneous melanoma (SKCM), ovarian serous cyst adenocarcinoma (OV), and prostate adenocarcinoma (PRAD). Further analysis comparing the gene expression changes between Ad.hPNPaseold‐35 infected HO‐1 melanoma cells and HeLa cells overexpressing hPNPaseold‐35 under the control of a doxycycline‐inducible promoter, revealed global changes in genes involved in cell cycle and mitosis. Overall, this study provides further evidence that hPNPaseold‐35 is associated with global changes in cell cycle‐associated genes and identifies potential gene targets for future investigation. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 14, 2014   doi: 10.1002/jcp.24645   open full text
• CCL27 Is Downregulated by Interferon Gamma via Epidermal Growth Factor Receptor in Normal Human Epidermal Keratinocytes.
  Masaru Karakawa, Mayumi Komine, Yasushi Hanakawa, Hidetoshi Tsuda, Koji Sayama, Kunihiko Tamaki, Mamitaro Ohtsuki.
  Journal of Cellular Physiology. April 08, 2014
  The cutaneous T cell‐attracting chemokine (CTACK)/CCL27 is indispensable in skin inflammation. CTACK/CCL27 is exclusively produced by epidermal keratinocytes to attract CCR10‐expressing T lymphocytes to the skin. We investigated the mechanism of CTACK/CCL27 production from normal human epidermal keratinocytes (NHEKs) by the proinflammatory cytokines TNFα and IFNγ. CTACK/CCL27 production was induced by TNFα via ERK, JNK, p38, and NFκB. The induction of CTACK/CCL27 by TNFα was suppressed by IFNγ via a pathway dependent on JAK, STAT1, and STAT3. Our results also demonstrated that IFNγ and TNFα induced the phosphorylation of EGFR and the following phosphorylation of ERK, which is partly responsible for the suppressive effect of IFNγ on TNFα‐induced production of CTACK/CCL27. Peri‐lesional skin of psoriasis demonstrates early inflammatory changes as we have previously reported. CTACK/CCL27 expression was diffuse in the peri‐lesional epidermis, while it was restricted to basal layer in lesional epidermis, suggesting that CTACK/CCL27 expression was induced in the early stage of psoriatic plaque formation, and IFNγ could participate in the suppression of CTACK/CCL27 expression in the lesional epidermis, reflecting the later stage of psoriatic plaque formation. Our study suggests that CTACK/CCL27 may have a pivotal role in the early stage of psoriasis plaque formation, but should be downregulated in the later stage to induce inflammation characteristic for chronic psoriasis plaques. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 08, 2014   doi: 10.1002/jcp.24643   open full text
• Inactivation of the LOX‐1 pathway promotes the Golgi apparatus during cell differentiation of mural granulosa cells.
  J.M. Weitzel, A. Vernunft, B. Krüger, C. Plinski, T. Viergutz.
  Journal of Cellular Physiology. April 08, 2014
  In female mammals, granulosa cells of the ovarian follicle differentiate into the corpus luteum after ovulation of the pregnable oocyte into the fallopian tube. During these differentiation processes several morphological alterations have to occur and the molecular basis is not fully understood. As an endpoint estradiol production from granulosa cells has to switch off in favor for progesterone production from the proceeding corpus luteum to sustain the developing embryo. Previously, we demonstrated that the multiligand receptor LOX‐1 plays a critical role in steroid hormone synthesis of granulosa cells via intracellular calcium release from endoplasmic (ER)‐dependent and ER‐independent calcium pools. In the present study, we show that inhibition of LOX‐1 leads to a rearrangement of ceramide from the basal membrane toward the Golgi apparatus. This activity is accomplished by a calcium‐dependent phosphorylation of aromatase, the key step in estradiol production. Phosphorylated aromatase increased estradiol production in a dose‐dependent manner. Our data indicate that the ceramide cascade is essential for proper granulosa cell function and ceramide redistribution serves as a first step in order to proceed with the prosperous differentiation into a corpus luteum. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 08, 2014   doi: 10.1002/jcp.24644   open full text
• Hypomethylating Agent 5‐Aza‐2′‐deoxycytidine (DAC) Ameliorates Multiple Sclerosis in Mouse Models.
  Katia Mangano, Paolo Fagone, Klaus Bendtzen, Pier Luigi Meroni, Cinzia Quattrocchi, Santa Mammana, Michelino Di Rosa, Lucia Malaguarnera, Marinella Coco, Gaetano Magro, Roberto Di Marco, Ferdinando Nicoletti.
  Journal of Cellular Physiology. April 02, 2014
  Increasing evidence supports the role of epigenetics in the development of autoimmune disorders and the possibility of using epigenetic modifying drugs in the context of MS has not yet been investigated. We have explored the effect of the hypomethylating agent 5‐aza‐2′‐deoxycytidine (DAC) in two murine models of experimental allergic encephalomyelitis (EAE). DAC treatment was associated with a significant amelioration of the clinical and histological hallmarks of EAE in both models. These effects were observed both in prophylactic and therapeutic regimens. The milder course of the disease was associated with a reduction in the number of spinal cord infiltrating lymphocytes and amelioration of the histopathological signs associated with EAE. In addition, increased transcript levels of anti‐inflammatory cytokines and decreased mRNA expression of pro‐inflammatory mediators were also observed. Finally, DAC treatment increased the percentage of circulating regulatory T cells by inducing Foxp3 expression via demethylation of a CpG island in Foxp3. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 02, 2014   doi: 10.1002/jcp.24641   open full text
• Primary cilium regulates CaV1.2 expression through Wnt signaling.
  Brian S. Muntean, Xingjian Jin, Frederick E. Williams, Surya M. Nauli.
  Journal of Cellular Physiology. April 02, 2014
  Primary cilia are sensory organelles that provide a feedback mechanism to restrict Wnt signaling in the absence of endogenous Wnt activators. Abnormal Wnt signaling has been shown to result in polycystic kidney disease (PKD) although the exact mechanism has been debated. Previously, we reported that the calcium channel CaV1.2 functions in primary cilia. In this study, we show that CaV1.2 expression level is regulated by Wnt signaling. This occurs through modulation of mitochondrial mass and activity resulting in increased reactive oxygen species which generate oxidative DNA lesions. We found that the subsequent cellular DNA damage response triggers increased CaV1.2 expression. In the absence of primary cilia where Wnt signaling is upregulated, we found that CaV1.2 is overexpressed as a compensatory mechanism. We show for the first time that CaV1.2 knockdown in zebrafish results in classic primary cilia defects including renal cyst formation, hydrocephalus, and left‐right asymmetry defects. Our study shows that suppressed Wnt signaling prevents CaV1.2 expression ultimately resulting in PKD phenotypes. Thus, CaV1.2 expression is tightly regulated through Wnt signaling and plays an essential sensory role in primary cilia necessary for cellular homeostasis. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  April 02, 2014   doi: 10.1002/jcp.24642   open full text
• Ang II–AT1R Increases Cell Migration Through PI3K/AKT and NF‐κB Pathways in Breast Cancer.
  Yanbin Zhao, Hongbin Wang, Xiuli Li, Mengru Cao, Hailing Lu, Qingwei Meng, Hui Pang, Hailin Li, Christina Nadolny, Xiaoqun Dong, Li Cai.
  Journal of Cellular Physiology. April 01, 2014
  Angiotensin II (Ang II), a biologically active peptide of the renin–angiotensin system (RAS), plays an important role in promoting cell migration via Angiotensin II type 1 receptor (AT1R). In this study, we examined the mechanisms by which Ang II affected cell migration in AT1R‐positive MDA‐MB‐231 human breast cancer cells. Ang II increased cell migration and expression of matrix metalloproteinase (MMP)‐2,‐9 in a dose‐dependent manner. Ang II‐mediated cell migration was reduced by specific blocking of MMP‐2 and MMP‐9, as well as with pretreatment with inhibitors of AT1R, phosphatidylinositol 3‐kinase (PI3K), Akt, and NF‐κB. Similarly, Ang II‐mediated expression of MMP‐2,‐9 was downregulated by pretreatment with inhibitors of AT1R and PI3K. In addition, Ang II treatment significantly induced phosphorylation of PI3K, Akt, and resulted in increased NF‐κB activity. These findings suggest that Ang II activates the AT1R/PI3K/Akt pathway, which further activates IKKα/β and NF‐κB, resulting in enhanced expression of MMP‐2,‐9 and migration in human breast cancer cells. Therefore, targeting Ang II/AT1R/PI3K/Akt/NF‐κB signaling could be a novel anti‐metastatic therapy for breast cancer. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 01, 2014   doi: 10.1002/jcp.24639   open full text
• Heregulin Negatively Regulates Transcription of ErbB2/3 Receptors via an AKT‐Mediated Pathway.
  Smita Awasthi, Anne W. Hamburger.
  Journal of Cellular Physiology. April 01, 2014
  Despite the importance of the ErbB2/3 heterodimer in breast cancer progression, the negative regulation of these receptors is still poorly understood. We demonstrate here for the first time that the ErbB3/4 ligand heregulin (HRG) reduced both ErbB2 and ErbB3 mRNA and protein levels in human breast cancer cell lines. In contrast, EGFR levels were unaffected by HRG treatment. The effect was rapid with a decline in steady‐state mRNA levels first noted 2 h after HRG treatment. HRG reduced the rate of transcription of ErbB2 and ErbB3 mRNA, but did not affect ErbB2 or ErbB3 mRNA stability. To test if ErbB2 kinase activity was required for the HRG‐induced downregulation, we treated cells with the ErbB2/EGFR inhibitor lapatinib. Lapatinib diminished the HRG‐induced decrease in ErbB2 and ErbB3 mRNA and protein, suggesting that the kinase activity of EGFR/ErbB2 is involved in the HRG‐induced receptor downregulation. Further, HRG‐mediated decreases in ErbB2/3 mRNA transcription are reversed by inhibiting the AKT but not MAPK pathway. To examine the functional consequences of HRG‐mediated decreases in ErbB receptor levels, we performed cell‐cycle analysis. HRG blocked cell‐cycle progression and lapatinib reversed this block. Our findings support a role for HRG in the negative regulation of ErbB expression and suggest that inhibition of ErbB2/3 signaling by ErbB2 directed therapies may interfere with this process. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 01, 2014   doi: 10.1002/jcp.24637   open full text
• Combined EGFR and Autophagy Modulation Impairs Cell Migration and Enhances Radiosensitivity in Human Glioblastoma Cells.
  Silvia Palumbo, Paolo Tini, Marzia Toscano, Giulia Allavena, Francesca Angeletti, Federico Manai, Clelia Miracco, Sergio Comincini, Luigi Pirtoli.
  Journal of Cellular Physiology. April 01, 2014
  Glioblastoma (GBM) remains the most aggressive and lethal brain tumor due to its molecular heterogeneity and high motility and invasion capabilities of its cells, resulting in high resistance to current standard treatments (surgery, followed by ionizing radiation combined with Temozolomide chemotherapy administration). Locus amplification, gene overexpression, and genetic mutations of epidermal growth factor receptor (EGFR) are hallmarks of GBM that can ectopically activate downstream signaling oncogenic cascades such as PI3K/Akt/mTOR pathway. Importantly, alteration of this pathway, involved also in the regulation of autophagy process, can improve radioresistance in GBM cells, thus promoting the aggressive phenotype of this tumor. In this work, the endogenous EGFR expression profile and autophagy were modulated to increase radiosensitivity behavior of human T98G and U373MG GBM cells. Our results primarily indicated that EGFR interfering induced radiosensitivity according to a decrease of the clonogenic capability of the investigated cells, and an effective reduction of the in vitro migratory features. Moreover, EGFR interfering resulted in an increase of Temozolomide (TMZ) cytotoxicity in T98G TMZ‐resistant cells. In order to elucidate the involvement of the autophagy process as pro‐death or pro‐survival role in cells subjected to EGFR interfering, the key autophagic gene ATG7 was silenced, thereby producing a transient block of the autophagy process. This autophagy inhibition rescued clonogenic capability of irradiated and EGFR‐silenced T98G cells, suggesting a pro‐death autophagy contribution. To further confirm the functional interplay between EGFR and autophagy pathways, Rapamycin‐mediated autophagy induction during EGFR modulation promoted further impairment of irradiated cells, in terms of clonogenic and migration capabilities. Taken together, these results might suggest a novel combined EGFR‐autophagy modulation strategy, to overcome intrinsic GBM radioresistance, thus improving the efficacy of standard treatments. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 01, 2014   doi: 10.1002/jcp.24640   open full text
• Role of Caspase‐3 Cleaved IP3R1 on Ca2+ Homeostasis and Developmental Competence of Mouse Oocytes and Eggs.
  Nan Zhang, Rafael A. Fissore.
  Journal of Cellular Physiology. April 01, 2014
  Apoptosis in most cell types is accompanied by altered Ca2+ homeostasis. During apoptosis, caspase‐3 mediated cleavage of the type 1 inositol 1,4,5‐trisphosphate receptor (IP3R1) generates a 95‐kDa C‐terminal fragment (C‐IP3R1), which represents the channel domain of the receptor. Aged mouse eggs display abnormal Ca2+ homeostasis and express C‐IP3R1, although whether or not C‐IP3R1 expression contributes to Ca2+ misregulation or a decrease in developmental competency is unknown. We sought to answer these questions by injecting in mouse oocytes and eggs cRNAs encoding C‐IP3R1. We found that: (1) expression of C‐IP3R1 in eggs lowered the Ca2+ content of the endoplasmic reticulum (ER), although, as C‐IP3R1 is quickly degraded at this stage, its expression did not impair pre‐implantation embryo development; (2) expression of C‐IP3R1 in eggs enhanced fragmentation associated with aging; (3) endogenous IP3R1 is required for aging associated apoptosis, as its down‐regulation prevented fragmentation, and expression of C‐IP3R1 in eggs with downregulated IP3R1 partly restored fragmentation; (4) C‐IP3R1 expression in GV oocytes resulted in persistent levels of protein, which abolished the increase in the ER releasable Ca2+ pool that occurs during maturation, undermined the Ca2+ oscillatory ability of matured eggs and their activation potential. Collectively, this study supports a role for IP3R1 and C‐IP3R1 in regulating Ca2+ homeostasis and the ER Ca2+ content during oocyte maturation. Nevertheless, the role of C‐IP3R1 on Ca2+ homeostasis in aged eggs seems minor, as in MII eggs the majority of endogenous IP3R1 remains intact and C‐IP3R1 undergoes rapid turnover. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  April 01, 2014   doi: 10.1002/jcp.24638   open full text
• Biological Effects of Insulin and Its Analogs on Cancer Cells With Different Insulin Family Receptor Expression.
  Laura Sciacca, Maria Francesca Cassarino, Marco Genua, Paolo Vigneri, Maria Giovanna Pennisi, Pasqualino Malandrino, Sebastiano Squatrito, Vincenzo Pezzino, Riccardo Vigneri.
  Journal of Cellular Physiology. March 28, 2014
  Hyperinsulinemia is a likely cause of the increased cancer incidence and mortality in diabetic patients, but its role is difficult to define in vivo. Previous in vitro studies testing the mitogenic potential of insulin and its analogs provided incomplete and sometimes contradictory results. To better evaluate cancer cell responsiveness to insulin, to its analogs and to IGF‐I, we measured under identical experimental conditions cell proliferation, invasiveness, and foci formation in six cancer cell lines with different insulin receptor family expression levels. The cancer cells studied have a different expression of insulin receptor (IR), its isoforms (IR‐A and IR‐B), and of the IGF‐I receptor. The data indicate that insulin stimulates proliferation in all cancer cell lines, invasiveness in some, and foci formation in none. Cancer cell responses to insulin (and IGF‐I) are not related to receptor expression levels; moreover, hormone‐stimulated proliferation and invasiveness are not correlated. IGF‐I is a more potent stimulator than insulin in most but not all cancer cell lines. Insulin analogs including M1 and M2 Glargine metabolites stimulate cancer cells similar to insulin. However, exceptions occur for specific analogs in particular cancer cells. In conclusion, in vitro insulin is an effective growth factor for all cancer cells but the biological response to insulin cannot be predicted on the basis of receptor expression levels. In the clinical setting, these observations should be taken in account when deciding treatment for diabetic patients who are at risk of undiscovered cancer or survivors of oncological diseases. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  March 28, 2014   doi: 10.1002/jcp.24635   open full text
• Probiotic L. reuteri Treatment Prevents Bone Loss in a Menopausal Ovariectomized Mouse Model.
  Robert A. Britton, Regina Irwin, Darin Quach, Laura Schaefer, Jing Zhang, Taehyung Lee, Narayanan Parameswaran, Laura R. McCabe.
  Journal of Cellular Physiology. March 27, 2014
  Estrogen deficiency is a major risk factor for osteoporosis that is associated with bone inflammation and resorption. Half of women over the age of 50 will experience an osteoporosis related fracture in their lifetime, thus novel therapies are needed to combat post‐menopausal bone loss. Recent studies suggest an important role for gut‐bone signaling pathways and the microbiota in regulating bone health. Given that the bacterium Lactobacillus reuteri ATCC PTA 6475 (L. reuteri) secretes beneficial immunomodulatory factors, we examined if this candidate probiotic could reduce bone loss associated with estrogen deficiency in an ovariectomized (Ovx) mouse menopausal model. Strikingly, L. reuteri treatment significantly protected Ovx mice from bone loss. Osteoclast bone resorption markers and activators (Trap5 and RANKL) as well as osteoclastogenesis are significantly decreased in L. reuteri treated mice. Consistent with this, L. reuteri suppressed Ovx‐induced increases in bone marrow CD4+ T‐lymphocytes (which promote osteoclastogenesis) and directly suppressed osteoclastogenesis in vitro. We also identified that L. reuteri treatment modifies microbial communities in the Ovx mouse gut. Together, our studies demonstrate that L. reuteri treatment suppresses bone resorption and loss associated with estrogen deficiency. Thus, L. reuteri treatment may be a straightforward and cost‐effective approach to reduce post‐menopausal bone loss. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  March 27, 2014   doi: 10.1002/jcp.24636   open full text
• Autophagy Is Modulated in Human Neuroblastoma Cells Through Direct Exposition to Low Frequency Electromagnetic Fields.
  Nicoletta Marchesi, Cecilia Osera, Lorenzo Fassina, Marialaura Amadio, Francesca Angeletti, Martina Morini, Giovanni Magenes, Letizia Venturini, Marco Biggiogera, Giovanni Ricevuti, Stefano Govoni, Salvatore Caorsi, Alessia Pascale, Sergio Comincini.
  Journal of Cellular Physiology. March 27, 2014
  In neurogenerative diseases, comprising Alzheimer's (AD), functional alteration in autophagy is considered one of the pathological hallmarks and a promising therapeutic target. Epidemiological investigations on the possible causes undergoing these diseases have suggested that electromagnetic fields (EMF) exposition can contribute to their etiology. On the other hand, EMF have therapeutic implications in reactivating neuronal functionality. To partly clarify this dualism, the effect of low‐frequency EMF (LF‐EMF) on the modulation of autophagy was investigated in human neuroblastoma SH‐SY5Y cells, which were also subsequently exposed to Aβ peptides, key players in AD. The results primarily point that LF‐EMF induce a significant reduction of microRNA 30a (miR‐30a) expression with a concomitant increase of Beclin1 transcript (BECN1) and its corresponding protein. Furthermore, LF‐EMF counteract the induced miR‐30a up‐regulation in the same cells transfected with miR‐30a mimic precursor molecules and, on the other side, rescue Beclin1 expression after BECN1 siRNA treatment. The expression of autophagy‐related markers (ATG7 and LC3B‐II) as well as the dynamics of autophagosome formation were also visualized after LF‐EMF exposition. Finally, different protocols of repeated LF‐EMF treatments were assayed to contrast the effects of Aβ peptides in vitro administration. Overall, this research demonstrates, for the first time, that specific LF‐EMF treatments can modulate in vitro the expression of a microRNA sequence, which in turn affects autophagy via Beclin1 expression. Taking into account the pivotal role of autophagy in the clearance of protein aggregates within the cells, our results indicate a potential cytoprotective effect exerted by LF‐EMF in neurodegenerative diseases such as AD. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  March 27, 2014   doi: 10.1002/jcp.24631   open full text
• Sox18 preserves the pulmonary endothelial barrier under conditions of increased shear stress.
  Christine M. Gross, Saurabh Aggarwal, Sanjiv Kumar, Jing Tian, Anita Kasa, Natalia Bogatcheva, Sanjeev A. Datar, Alexander D. Verin, Jeffrey R. Fineman, Stephen M. Black.
  Journal of Cellular Physiology. March 27, 2014
  Shear stress secondary to increased pulmonary blood flow (PBF) is elevated in some children born with congenital cardiac abnormalities. However, the majority of these patients do not develop pulmonary edema, despite high levels of permeability inducing factors. Previous studies have suggested that laminar fluid shear stress can enhance pulmonary vascular barrier integrity. However, little is known about the mechanisms by which this occurs. Using microarray analysis, we have previously shown that Sox18, a transcription factor involved in blood vessel development and endothelial barrier integrity, is up‐regulated in an ovine model of congenital heart disease with increased PBF (shunt). By subjecting ovine pulmonary arterial endothelial cells (PAEC) to laminar flow (20 dyn/cm2), we identified an increase in trans‐endothelial resistance (TER) across the PAEC monolayer that correlated with an increase in Sox18 expression. Further, the TER was also enhanced when Sox18 was over‐expressed and attenuated when Sox18 expression was reduced, suggesting that Sox18 maintains the endothelial barrier integrity in response to shear stress. Further, we found that shear stress up‐regulates the cellular tight junction protein, Claudin‐5, in a Sox18 dependent manner, and Claudin‐5 depletion abolished the Sox18 mediated increase in TER in response to shear stress. Finally, utilizing peripheral lung tissue of 4 week old shunt lambs with increased PBF, we found that both Sox18 and Claudin‐5 mRNA and protein levels were elevated. In conclusion, these novel findings suggest that increased laminar flow protects endothelial barrier function via Sox18 dependent up‐regulation of Claudin‐5 expression.

  March 27, 2014   doi: 10.1002/jcp.24633   open full text
• Functional Inhibition of Aquaporin‐3 With a Gold‐Based Compound Induces Blockage of Cell Proliferation.
  Ana Serna, Ana Galán‐Cobo, Claudia Rodrigues, Ismael Sánchez‐Gomar, Juan José Toledo‐Aral, Teresa F. Moura, Angela Casini, Graça Soveral, Miriam Echevarría.
  Journal of Cellular Physiology. March 27, 2014
  AQP3 has been correlated with higher transport of glycerol, increment of ATP content, and larger proliferation capacity. Recently, we described the gold(III) complex Auphen as a very selective and potent inhibitor of AQP3's glycerol permeability (Pgly). Here we evaluated Auphen effect on the proliferation of various mammalian cell lines differing in AQP3 expression level: no expression (PC12), moderate (NIH/3T3) or high (A431) endogenous expression, cells stably expressing AQP3 (PC12‐AQP3), and human HEK293T cells transiently transfected (HEK‐AQP3) for AQP3 expression. Proliferation was evaluated in the absence or presence of Auphen (5 μM) by counting number of viable cells and analyzing 5‐bromo‐2′‐deoxyuridine (BrdU) incorporation. Auphen reduced ≈50% the proliferation in A431 and PC12‐AQP3, ≈15% in HEK‐AQP3 and had no effect in wt‐PC12 and NIH/3T3. Strong arrest in the S‐G2/M phases of the cell cycle, supported by analysis of cyclins (A, B1, D1, E) levels, was observed in AQP3‐expressing cells treated with Auphen. Flow‐cytometry of propidium iodide incorporation and measurements of mitochondrial dehydrogenases activity confirmed absence of cytotoxic effect of the drug. Functional studies evidenced ≈50% inhibition of A431 Pgly by Auphen, showing that the compound's anti‐proliferative effect correlates with its ability to inhibit AQP3 Pgly. Role of Cys‐40 on AQP3 permeability blockage by Auphen was confirmed by analyzing the mutated protein (AQP3‐Ser‐40). Accordingly, cells transfected with mutated AQP3 gained resistance to the antiproliferative effect of Auphen. These results highlight an Auphen inhibitory effect on proliferation of cells expressing AQP3 and suggest a targeted therapeutic effect on carcinomas with large AQP3 expression. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  March 27, 2014   doi: 10.1002/jcp.24632   open full text
• Osteoactivin Induces Transdifferentiation of C2C12 Myoblasts Into Osteoblasts.
  Gregory R. Sondag, Sibel Salihoglu, Suzanne L. Lababidi, Douglas C. Crowder, Fouad M. Moussa, Samir M. Abdelmagid, Fayez F. Safadi.
  Journal of Cellular Physiology. March 21, 2014
  Osteoactivin (OA) is a novel osteogenic factor important for osteoblast differentiation and function. Previous studies showed that OA stimulates matrix mineralization and transcription of osteoblast specific genes required for differentiation. OA plays a role in wound healing and its expression was shown to increase in post fracture calluses. OA expression was reported in muscle as OA is upregulated in cases of denervation and unloading stress. The regulatory mechanisms of OA in muscle and bone have not yet been determined. In this study, we examined whether OA plays a role in transdifferentiation of C2C12 myoblast into osteoblasts. Infected C2C12 with a retroviral vector overexpressing OA under the CMV promoter were able to transdifferentiate from myoblasts into osteoblasts. Immunofluorescence analysis showed that skeletal muscle marker MF‐20 was severely downregulated in cells overexpressing OA and contained significantly less myotubes compared to uninfected control. C2C12 myoblasts overexpressing OA showed an increase in expression of bone specific markers such as alkaline phosphatase and alizarin red staining, and also showed an increase in Runx2 protein expression. We also detected increased levels of phosphorylated focal adhesion kinase (FAK) in C2C12 myoblasts overexpressing OA compared to control. Taken together, our results suggest that OA is able to induce transdifferentiation of myoblasts into osteoblasts through increasing levels of phosphorylated FAK. J. Cell. Physiol. 229: 955–966, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24512   open full text
• BMP‐2 Induction of Dlx3 Expression Is Mediated by p38/Smad5 Signaling Pathway in Osteoblastic MC3T3‐E1 Cells.
  Guobin Yang, Guohua Yuan, Xiaoyan Li, Pingxian Liu, Zhi Chen, Mingwen Fan.
  Journal of Cellular Physiology. March 21, 2014
  Dlx3 is essential for osteoblast differentiation and bone formation, and its expression is regulated by bone morphogenetic protein‐2 (BMP‐2). However, the intimate mechanism of BMP‐2 regulation of Dlx3 transcription in osteoblasts is still unknown. Considering the important roles of Smad5 and p38 in osteoblast differentiation, we hypothesized that Smad5 and p38 mediated BMP‐2‐induced Dlx3 transcription in osteoblasts. We found activation of Smad5 and p38 increased the expression of Dlx3, whereas knocking down Smad5 or inactivation of p38 inhibited BMP‐2‐induced Dlx3 expression. Both Smad5 and p38 were able to activate Dlx3 promoter activity and p38/Smad5 response elements were located from −698 to −368 in Dlx3 promoter. Two Smad5 binding sites (SBEI and SBEII, TGTCT box) were identified in this region by EMSA and ChIP assay. Deletions and mutagenesis study of the Dlx3 promoter region indicated that the TGTCT boxes are crucial for p38/Smad5‐induced Dlx3 promoter activity. At last, we found a cross‐talk between p38 and Smad5, and that activation of p38 is necessary for BMP‐2‐induced Smad5 phosphorylation and nuclear translocation. Overall, we provide a novel insight that BMP‐2‐induced Dlx3 expression is regulated by p38/Smad5 signaling pathway in osteoblasts. J. Cell. Physiol. 229: 943–954, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24525   open full text
• Regulation of Plasticity and Fibrogenic Activity of Trabecular Meshwork Cells by Rho GTPase Signaling.
  Padmanabhan P. Pattabiraman, Rupalatha Maddala, Ponugoti Vasantha Rao.
  Journal of Cellular Physiology. March 21, 2014
  Glaucoma, a prevalent blinding disease is commonly associated with increased intraocular pressure due to impaired aqueous humor (AH) drainage through the trabecular meshwork (TM). Although increased TM tissue contraction and stiffness in association with accumulation of extracellular matrix (ECM) are believed to be partly responsible for increased resistance to AH outflow, the extracellular cues and intracellular mechanisms regulating TM cell contraction and ECM production are not well defined. This study tested the hypothesis that sustained activation of Rho GTPase signaling induced by lysophosphatidic acid (LPA), TGF‐β, and connective tissue growth factor (CTGF) influences TM cell plasticity and fibrogenic activity which may eventually impact resistance to AH outflow. Various experiments performed using human TM cells revealed that constitutively active RhoA (RhoAV14), TGF‐β2, LPA, and CTGF significantly increase the levels and expression of Fibroblast Specific Protein‐1 (FSP‐1), α‐smooth muscle actin (αSMA), collagen‐1A1 and secretory total collagen, as determined by q‐RT‐PCR, immunofluorescence, immunoblot, flow cytometry and the Sircol assay. Significantly, these changes appear to be mediated by Serum Response Factor (SRF), myocardin‐related transcription factor (MRTF‐A), Slug, and Twist‐1, which are transcriptional regulators known to control cell plasticity, myofibroblast generation/activation and fibrogenic activity. Additionally, the Rho kinase inhibitor‐Y27632 and anti‐fibrotic agent‐pirfenidone were both found to suppress the TGF‐β2‐induced expression of αSMA, FSP‐1, and collagen‐1A1. Taken together, these observations demonstrate the significance of RhoA/Rho kinase signaling in regulation of TM cell plasticity, fibrogenic activity, and myofibroblast activation, events with potential implications for the pathobiology of elevated intraocular pressure in glaucoma patients. J. Cell. Physiol. 229: 927–942, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24524   open full text
• Krüppel‐Like Factor 4 Regulates Blood‐Tumor Barrier Permeability via ZO‐1, Occludin and Claudin‐5.
  Jun Ma, Ping Wang, Yunhui Liu, Lini Zhao, Zhen Li, Yixue Xue.
  Journal of Cellular Physiology. March 21, 2014
  Blood‐tumor barrier (BTB) constitutes an efficient organization of tight junctions which significantly reduce permeability for chemotherapy drugs. Krüppel‐like factor 4 (KLF4), a member of the Krüppel‐like family, has been documented in endothelial cells and may serve as an essential regulator of endothelial barrier function. However, our knowledge about the expression and function of KLF4 in the endothelial cells of BTB still remains unclear. In this study, we sought to investigate the role of KLF4 in regulation of BTB function as well as the potential molecular mechanisms. Quantitative RT‐PCR, Western blot, and immunofluorescence assays demonstrated that KLF4 was down‐regulated in the glioma endothelial cells (GECs) which were obtained through endothelial cells co‐cultured with glioma cells. Short hairpin RNA targeting KLF4 impaired the integrity of BTB detected by trans‐endothelial electric resistance assay, and meanwhile reduced the expression of ZO‐1, occludin and claudin‐5, demonstrated by quantitative RT‐PCR, Western blot, and immunofluorescence assays. Depletion of KLF4 increased BTB permeability to small molecules detected by permeability assays. Furthermore, luciferase assays and chromatin immunoprecipitation assays showed that KLF4 up‐regulated the promoter activities and interacted with “CACCC” DNA sequence presented in the promoters of ZO‐1, occludin, and claudin‐5. GATA‐1, GATA‐6, Sp1, and Sp3 factors participated in KLF4 regulation of promoter activities through binding to the promoters of tight junctions related proteins. Collectively, our results indicated that KLF4 is a key transcriptional regulator of BTB function by regulating expressions of tight junction related proteins, which would draw growing attention to KLF4 as a potential target for glioma therapy. J. Cell. Physiol. 229: 916–926, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24523   open full text
• MEK1/2 Overactivation Can Promote Growth Arrest by Mediating ERK1/2‐Dependent Phosphorylation of p70S6K.
  Jean‐Philippe Guégan, Frédéric Ezan, Luc Gailhouste, Sophie Langouët, Georges Baffet.
  Journal of Cellular Physiology. March 21, 2014
  The extracellular signal‐regulated kinase (ERK)1/2 mitogen‐activated protein (MAP) kinase pathway has been involved in the positive and negative regulation of cell proliferation. Upon mitogen stimulation, ERK1/ERK2 activation is necessary for G1‐ to S‐phase progression whereas when hyperactived, this pathway could elicit cell cycle arrest. The mechanisms involved are not fully elucidated but a kinase‐independent function of ERK1/2 has been evidenced in the MAPK‐induced growth arrest. Here, we show that p70S6K, a central regulator of protein biosynthesis, is essential for the cell cycle arrest induced by overactivation of ERK1/2. Indeed, whereas MEK1 silencing inhibits cell cycle progression, we demonstrate that active mutant form of MEK1 or MEK2 triggers a G1 phase arrest by stimulating an activation of p70S6K by ERK1/2 kinases. Silencing of ERK1/2 activity by shRNA efficiently suppresses p70S6K phosphorylation on Thr421/Ser424 and S6 phosphorylation on Ser240/244 as well as p21 expression, but these effects can be partially reversed by the expression of kinase‐dead mutant form of ERK1 or ERK2. In addition, we demonstrate that the kinase p70S6K modulates neither the p21 gene transcription nor the stability of the protein but enhances the translation of the p21 mRNA. In conclusion, our data emphasizes the importance of the translational regulation of p21 by the MEK1/2‐ERK1/2‐p70S6K pathway to negatively control the cell cycle progression. J. Cell. Physiol. 229: 903–915, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24521   open full text
• Metabolomics Biomarkers of Frailty in Elderly Breast Cancer Patients.
  Giuseppe Corona, Jerry Polesel, Lucia Fratino, Gianmaria Miolo, Flavio Rizzolio, Diana Crivellari, Riccardo Addobbati, Silvia Cervo, Giuseppe Toffoli.
  Journal of Cellular Physiology. March 21, 2014
  Metabolome analysis has emerged as a powerful technique for detecting and define specific physio‐pathological phenotypes. In this investigation the diagnostic potential of metabolomics has been applied to better characterize the multiple biochemical alterations that concur in the definition of the frailty phenotype observed in elderly breast cancer patients. The study included 89 women with breast cancer (range 70–97 years) classified as Fit (n = 49), Unfit (n = 23), or Frail (n = 17) according to comprehensive geriatric assessment. The serum metabolomic profile was performed by tandem mass spectrometry and included different classes of metabolites such as amino acids, acylcarnitines, sphingo‐, and glycerol‐phospolipids. ANOVA was applied to identify the metabolites differing significantly among Fit, Unfit, and Frail patients. In patients carrying the frail phenotype, the amino acid perturbations involve serine, tryptophan, hydroxyproline, histidine, its derivate 3‐methyl‐hystidine, cystine, and β‐aminoisobutyric acid. With regard to lipid metabolism, the frailty phenotype was characterized by a decrease of a wide number of glycerol‐ and sphingo‐phospholipid metabolites. These metabolomics biomarkers may give a further insight into the biochemical processes involved in the development of frailty in breast cancer patients. Moreover, they might be useful to refine the comprehensive geriatric assessment model. J. Cell. Physiol. 229: 898–902, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24520   open full text
• Overexpression of Calreticulin Contributes to the Development and Progression of Pancreatic Cancer.
  Weiwei Sheng, Chuanping Chen, Ming Dong, Jianping Zhou, Qingfeng Liu, Qi Dong, Feng Li.
  Journal of Cellular Physiology. March 21, 2014
  We studied the clinicopathological significance for Calreticulin (CRT) expression in pancreatic cancer (PC), and its functional relationship with other signaling genes (especially with p53) in regulating the biological behavior of PC cells. IHC, IF, IB, and real‐time PCR were used to detect CRT expression in PC, while transfection and drug intervention were used to investigate the functional relationship of CRT with other signaling genes. IHC showed both CRT and p53 expression was significantly increased in PC, compared to that in paired non‐cancerous pancreatic tissues (P < 0.001). High expression of CRT was positively associated with tumor UICC stage and lymph nodes metastasis (P = 0.034 and P = 0.015), and was an independent adverse prognostic indicator in patients with PC. No relationship was found between CRT and p53 expression in spearman's rank correlation test. Altered expression of CRT did not change p53, MDM2, pho‐AKT, pho‐p38, and pho‐JNK expression, but had a specific regulation on pho‐ERK. Meanwhile, CRT‐regulated cell proliferation, migration, and invasion of PC cells in MEK/ERK pathway dependent manner. In addition, CRT knockdown significantly decreased pho‐ERK expression and cell chemoresistance independent of activated p53 and caspase‐3‐related apoptosis in gemcitabine‐ or oxaliplatin‐treated Capan‐2 cells. Our study first demonstrated that overexpression of CRT contributed to the development and progression of PC through MEK/ERK‐signaling pathway but independent of p53. The interaction between CRT and MEK/ERK pathway might provide a new idea for revealing malignant biology and supplying new gene targeted chemotherapy of PC. J. Cell. Physiol. 229: 887–897, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24519   open full text
• Effect of Mechanical Strain on the Collagen VI Pericellular Matrix in Anterior Cruciate Ligament Fibroblasts.
  Francesca Sardone, Francesco Traina, Francesca Tagliavini, Camilla Pellegrini, Luciano Merlini, Stefano Squarzoni, Spartaco Santi, Simona Neri, Cesare Faldini, Nadir Maraldi, Patrizia Sabatelli.
  Journal of Cellular Physiology. March 21, 2014
  Cell–extracellular matrix interaction plays a major role in maintaining the structural integrity of connective tissues and sensing changes in the biomechanical environment of cells. Collagen VI is a widely expressed non‐fibrillar collagen, which regulates tissues homeostasis. The objective of the present investigation was to extend our understanding of the role of collagen VI in human ACL. This study shows that collagen VI is associated both in vivo and in vitro to the cell membrane of knee ACL fibroblasts, contributing to the constitution of a microfibrillar pericellular matrix. In cultured cells the localization of collagen VI at the cell surface correlated with the expression of NG2 proteoglycan, a major collagen VI receptor. The treatment of ACL fibroblasts with anti‐NG2 antibody abolished the localization of collagen VI indicating that collagen VI pericellular matrix organization in ACL fibroblasts is mainly mediated by NG2 proteoglycan. In vitro mechanical strain injury dramatically reduced the NG2 proteoglycan protein level, impaired the association of collagen VI to the cell surface, and promoted cell cycle withdrawal. Our data suggest that the injury‐induced alteration of specific cell–ECM interactions may lead to a defective fibroblast self‐renewal and contribute to the poor regenerative ability of ACL fibroblasts. J. Cell. Physiol. 229: 878–886, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24518   open full text
• The ZnR/GPR39 Interacts With the CaSR to Enhance Signaling in Prostate and Salivary Epithelia.
  Hila Asraf, Shimrit Salomon, Andrey Nevo, Israel Sekler, Doris Mayer, Michal Hershfinkel.
  Journal of Cellular Physiology. March 21, 2014
  Zinc signaling is mediated by the zinc sensing receptor, ZnR, recently suggested to be the same receptor as G‐protein coupled receptor 39, GPR39. However, it is unknown if GPR39 is mediating Zn2+‐dependent signaling in prostate and salivary tissue where changes in zinc concentrations are frequent and of physiological significance. Here, we show that GPR39 is mediating Zn2+‐dependent Ca2+ responses and is regulating activity of MAP and PI3 pathways in prostate cancer cells, PC3, and ductal salivary gland cells, HSY. We next ask whether ZnR/GPR39 interacts with other GPCR family members. We find that endogenous ZnR/GPR39 activity is regulated by the expression and activity of another cation sensing GPCR, the Ca2+‐sensing receptor (CaSR). Although CaSR is not activated by Zn2+, co‐expression of CaSR and ZnR/GPR39 synergistically enhances Ca2+ responses in PC3 and HSY cells. Silencing of the CaSR using siRNA or a dominant negative construct reduces the Zn2+‐dependent signaling. Importantly, overexpression of GPR39 in HEK293 cells is sufficient to trigger Zn2+‐dependent responses. Nevertheless, application of the CaSR agonist spermine, at concentration below its threshold, enhanced Zn2+‐dependent Ca2+ response. Our results suggest that the CaSR interacts with ZnR/GPR39 and thereby regulates its activity. Finally, we show that in PC3 cells ZnR/GPR39 is required for mediating the Zn2+‐dependent activation of MAPK and PI3K, pathways leading to enhanced cell growth. Importantly, Zn2+‐dependent activation of ZnR/GPR39 also enhances the expression of the Ca2+‐binding protein S100A4 that is linked to invasion of prostate cancer cells. J. Cell. Physiol. 229: 868–877, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24514   open full text
• ERK5 Pathway Regulates Transcription Factors Important for Monocytic Differentiation of Human Myeloid Leukemia Cells.
  Xuening Wang, Stella Pesakhov, Jonathan S. Harrison, Michael Danilenko, George P. Studzinski.
  Journal of Cellular Physiology. March 21, 2014
  Mitogen‐activated protein kinases (MAPKs) are important transducers of external signals for cell growth, survival, and other cellular responses including cell differentiation. Several MAPK cascades are known with the MEK1/2‐ERK1/2, JNK, and p38MAPKs receiving most attention, but the role of MEK5‐ERK5 in intracellular signaling deserves more scrutiny, as this pathway transmits signals that can complement ERK/2 signaling. We hypothesized that the ERK5 pathway plays a role in the control of monocytic differentiation, which is disturbed in myeloid leukemia. We therefore examined the cellular phenotype and key molecular events which occur when human myeloid leukemia cells, acute (AML) or chronic (CML), are forced to differentiate by vitamin D derivatives (VDDs). This study was performed using established cell lines HL60 and U937, and primary cultures of blasts from 10 patients with ML. We found that ERK5 and its direct downstream target transcription factor MEF2C are upregulated by 1,25D in parallel with monocytic differentiation. Further, inhibition of ERK5 activity by specific pharmacological agents BIX02189 and XMD8‐92 alters the phenotype of these cells by reducing the abundance of the VDD‐induced surface monocytic marker CD14, and concomitantly increasing surface expression of the general myeloid marker CD11b. Similar results were obtained when the expression of ERK5 was reduced by siRNA or short hairpin (sh) RNA. ERK5 inhibition resulted in an expected decrease in MEF2C activation. We also found that in AML cells the transcription factor C/EBPβ is positively regulated, while C/EBPα is negatively regulated by ERK5. These findings provide new understanding of dysregulated differentiation in human myeloid leukemia. J. Cell. Physiol. 229: 856–867, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24513   open full text
• Acetylsalicylic Acid Inhibits IL‐18‐Induced Cardiac Fibroblast Migration Through the Induction of RECK.
  Jalahalli M. Siddesha, Anthony J. Valente, Siva S.V.P. Sakamuri, Jason D. Gardner, Patrice Delafontaine, Makoto Noda, Bysani Chandrasekar.
  Journal of Cellular Physiology. March 21, 2014
  The pathogenesis of cardiac fibrosis and adverse remodeling is thought to involve the ROS‐dependent induction of inflammatory cytokines and matrix metalloproteinases (MMPs), and the activation and migration of cardiac fibroblasts (CF). Here we investigated the role of RECK (reversion‐inducing‐cysteine‐rich protein with Kazal motifs), a unique membrane‐anchored MMP regulator, on IL‐18‐induced CF migration, and the effect of acetylsalicylic acid (ASA) on this response. In a Matrigel invasion assay, IL‐18‐induced migration of primary mouse CF was dependent on both IKK/NF‐κB‐ and JNK/AP‐1‐mediated MMP9 induction and Sp1‐mediated RECK suppression, mechanisms that required Nox4‐dependent H2O2 generation. Notably, forced expression of RECK attenuated IL‐18‐induced MMP9 activation and CF migration. Further, therapeutic concentrations of ASA inhibited IL‐18‐induced H2O2 generation, MMP9 activation, RECK suppression, and CF migration. The salicylic acid moiety of ASA similarly attenuated IL‐18‐induced CF migration. Thus, ASA may exert potential beneficial effect in cardiac fibrosis through multiple protective mechanisms. J. Cell. Physiol. 229: 845–855, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24511   open full text
• Epigenetics and Ocular Diseases: From Basic Biology to Clinical Study.
  Biao Yan, Jin Yao, Zhi‐Fu Tao, Qin Jiang.
  Journal of Cellular Physiology. March 21, 2014
  Epigenetics is an emerging field in ophthalmology and has opened a new avenue for understanding ocular development and ocular diseases related to aging and environment. Epigenetic mechanisms, including DNA methylation, histone modifications, chromatin remodeling, and deployment of non‐coding RNAs, result in the heritable silencing of gene expression without any change in DNA sequence. Accumulating evidence suggests a potential link between gene expression, chromatin structure, non‐coding RNAs, and cellular differentiation during ocular development. Disruption of the balance of epigenetic networks could become the etiology of several ocular diseases. Here, we summarized the current knowledge about epigenetic regulatory mechanisms in ocular development and diseases. J. Cell. Physiol. 229: 825–833, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24522   open full text
• miR‐663 Induces Castration‐Resistant Prostate Cancer Transformation and Predicts Clinical Recurrence.
  Li Jiao, Zhen Deng, Chuanliang Xu, Yongwei Yu, Yun Li, Chun Yang, Junyi Chen, Zhiyong liu, Gang Huang, Long‐Cheng Li, Yinghao Sun.
  Journal of Cellular Physiology. March 21, 2014
  Castration‐resistant prostate cancer (CRPC) and its treatment are challenging issues in prostate cancer management. Here, we report that miR‐663 is upregulated in CRPC tissues. Overexpression of miR‐663 in prostate LNCaP cells promotes cell proliferation and invasion, neuroendocrine differentiation, and reduction in dihydrotestosterone‐induced upregulation of prostate‐specific antigen expression. Furthermore, results of in situ hybridization show that miR‐663 expression is correlated with Gleason score and TNM stage and is an independent prognostic predictor of clinical recurrence. Together, these findings suggest that miR‐663 is a potential oncomiR for CRPC and may serve as a tumor biomarker for the early diagnosis of CRPC. J. Cell. Physiol. 229: 834–844, 2014. © 2013 Wiley Periodicals, Inc.

  March 21, 2014   doi: 10.1002/jcp.24510   open full text
• RhoA‐Mediated Inhibition of Vascular Endothelial Cell Mobility: Positive Feedback Through Reduced Cytosolic p21 and p27.
  Yung‐Ho Hsu, Chih‐Cheng Chang, Nian‐Jie Yang, Yi‐Hsuan Lee, Shu‐Hui Juan.
  Journal of Cellular Physiology. February 18, 2014
  We previously identified that activation of the aryl hydrocarbon receptor (AhR) by 3‐methylcholanthrene (3MC) exerts antiproliferative and antimigratory effects on human umbilical vein endothelial cells (HUVECs) through the upregulation of p21/p27 transcription and RhoA activation. In this study, we investigated the mechanisms of 3MC‐mediated downregulation of cytosolic p21/ p27, and the effects of 3MC on RhoA activation and cell migration, in mouse cerebral vascular endothelial cells (MCVECs). Our results indicated that 3MC reduced the phosphorylation of p21/p27 through AhR/RhoA/PTEN‐mediated PI3K/Akt inactivation, which reduced cytosolic p21/p27 retention, causing RhoA activation through positive feedback. Downregulation of p21/p27 by siRNA, and cytosolic p21/p27 by the nuclear export blocker leptomycin B, further reduced cell migration in the 3MC‐treated cells. Reduced cytosolic p21/p27 expression led to reduced interaction between RhoA and the RhoA inhibitor p190RhoGAP, causing RhoA activation. Treatment with YS‐49 activated PI3K/Akt, a downstream target of RhoA, to reduce RhoA/PTEN activation in the 3MC‐treated cells, whereas treatment with wortmannin, a PI3K inhibitor, activated RhoA/PTEN. Gain‐ and loss‐of‐function analyses revealed that constitutively active (CA) Akt1, but not CA Akt2, inactivated RhoA and stimulated migratory activity. Considering the essential role of RhoA activation in cell migration, we evaluated the potential use of simvastatin, a RhoA inhibitor, as a therapeutic intervention in vivo using matrigel plug formation assays. Our results provide a molecular basis for the therapeutic application of simvastatin to reduce RhoA/PTEN activation, restore cytosolic levels of phosphorylated p21/p27, and induce angiogenic processes. J. Cell. Physiol. 9999: XX–XX, 2014. © 2014 Wiley Periodicals, Inc.

  February 18, 2014   doi: 10.1002/jcp.24583   open full text
• Pancreatic Cancer‐Induced Cachexia Is Jak2‐Dependent in Mice.
  Marine Gilabert, Ezequiel Calvo, Ana Airoldi, Tewfik Hamidi, Vincent Moutardier, Olivier Turrini, Juan Iovanna.
  Journal of Cellular Physiology. February 14, 2014
  Cancer cachexia syndrome is observed in 80% of patients with advanced‐stage cancer, and it is one of the most frequent causes of death. Severe wasting accounts for more than 80% in patients with advanced pancreatic cancer. Here we wanted to define, by using an microarray approach and the Pdx1‐cre;LSL‐KrasG12D;INK4a/arffl/fl mice model, the pathways involved in muscle, liver and white adipose tissue wasting. These mice, which develop systematically pancreatic cancer, successfully reproduced many human symptoms afflicted with this disease, and particularly cachexia. Using the profiling analysis of pancreatic cancer‐dependent cachectic tissues we found that Jak2/Stat3 pathways, p53 and NFkB results activated. Thus, our interest was focused on the Jak2 pathways because it is pharmacologically targetable with low toxicity and FDA approved drugs are available. Therefore, Pdx1‐cre;LSL‐KrasG12D;INK4a/arffl/fl mice were treated with the Jak2 inhibitor AG490 compound daily starting at 7 weeks‐old and for a period of 3 weeks and animals were sacrificed a 10 weeks old. Body weight for control mice was 27.84 ± 2.14 gr, for untreated Pdx1‐cre;LSL‐KrasG12D;INK4a/arffl/fl was 14.97 ± 1.99 gr, whereas in animals treated with the AG490 compound the weight loss was significantly less to 24.53 ± 2.04 gr. Treatment with AG490 compound was efficient since phosphorylation of Jak2 and circulating IL6 levels were significantly reduced in cachectic tissues and in mice respectively. In conclusion, we found that Jak2/Stat3‐dependent intracellular pathway plays an essential role since its pharmacological inhibition strongly attenuates cachexia progression in a lethal transgenic pancreatic cancer model. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 14, 2014   doi: 10.1002/jcp.24580   open full text
• Link Between DNA Damage and Centriole Disengagement/Reduplication in Untransformed Human Cells.
  Stephen Douthwright, Greenfield Sluder.
  Journal of Cellular Physiology. February 14, 2014
  The radiation and radiomimetic drugs used to treat human tumors damage DNA in both cancer cells and normal proliferating cells. Centrosome amplification after DNA damage is well established for transformed cell types but is sparsely reported and not fully understood in untransformed cells. We characterize centriole behavior after DNA damage in synchronized untransformed human cells. One hour treatment of S phase cells with the radiomimetic drug, Doxorubicin, prolongs G2 by at least 72 hours, though 14% of the cells eventually go through mitosis in that time. By 72 hours after DNA damage we observe a 52% incidence of centriole disengagement plus a 10% incidence of extra centrioles. We find that either APC/C or Plk activities can disengage centrioles after DNA damage, though they normally work in concert. All disengaged centrioles are associated with γ‐tubulin and maturation markers and thus, should in principle be capable of reduplicating and organizing spindle poles. The low incidence of reduplication of disengaged centrioles during G2 is due to the p53 dependent expression of p21 and the consequent loss of Cdk2 activity. We find that 26% of the cells going through mitosis after DNA damage contain disengaged or extra centrioles. This could produce genomic instability through transient or persistent spindle multipolarity. Thus, for cancer patients the use of DNA damaging therapies raises the chances of genomic instability and evolution of transformed characteristics in proliferating normal cell populations. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 14, 2014   doi: 10.1002/jcp.24579   open full text
• Adiponectin as Novel Regulator of Cell Proliferation in Human Glioblastoma.
  Carola Porcile, Erika Di Zazzo, Maria Ludovica Monaco, Giorgia D'Angelo, Daniela Passarella, Claudio Russo, Alfonso Di Costanzo, Alessandra Pattarozzi, Monica Gatti, Adriana Bajetto, Gianluigi Zona, Federica Barbieri, Giovannangelo Oriani, Bruno Moncharmont, Tullio Florio, Aurora Daniele.
  Journal of Cellular Physiology. February 14, 2014
  Adiponectin (Acrp30) is an adipocyte‐secreted hormone with pleiotropic metabolic effects, whose reduced levels were related to development and progression of several malignancies. We looked at the presence of Acrp30 receptors in human glioblastomas (GBM), hypothesizing a role for Acrp30 also in this untreatable cancer. Here we demonstrate that human GBM express Acrp30 receptors (AdipoR1 and AdipoR2), which are often co‐expressed in GBM samples (70% of the analyzed tumors). To investigate the effects of Acrp30 on GBM growth, we used human GBM cell lines U87‐MG and U251, expressing both AdipoR1 and AdipoR2 receptors. In these cells, Acrp30 treatment inhibits DNA synthesis and cell proliferation rate, inducing arrest in G1 phase of the cell cycle. These effects were correlated to a sustained activation of ERK1/2 and Akt kinases, upon Acrp30 treatment. Our results suggest that Acrp30 may represent a novel endogenous negative regulator of GBM cell proliferation, to be evaluated for the possible development of novel pharmacological approaches. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 14, 2014   doi: 10.1002/jcp.24582   open full text
• GRP78 Mediates Cell Growth and Invasiveness in Endometrial Cancer.
  Gaetano Calì, Luigi Insabato, Domenico Conza, Giuseppe Bifulco, Luca Parrillo, Paola Mirra, Francesca Fiory, Claudia Miele, Gregory Alexander Raciti, Bruno Di Jeso, Giuseppe Terrazzano, Francesco Beguinot, Luca Ulianich.
  Journal of Cellular Physiology. February 13, 2014
  Recent studies have indicated that endoplasmic reticulum stress, the unfolded protein response activation and altered GRP78 expression can play an important role in a variety of tumors development and progression. Very recently we reported for the first time that GRP78 is increased in endometrial tumors. However, whether GRP78 could play a role in the growth and/or invasiveness of endometrial cancer cells is still unknown. Here we report that the silencing of GRP78 expression affects both cell growth and invasiveness of Ishikawa and AN3CA cells, analyzed by the (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide) and transwell migration assay, respectively. At variance with Ishikawa cells, AN3CA cells showed, besides an endoplasmic reticulum, also a plasma membrane GRP78 localization, evidenced by both immunofluorescence and cell membrane biotinylation experiments. Intriguingly, flow cytometry experiments showed that the treatment with a specific antibody targeting GRP78 C‐terminal domain caused apoptosis in AN3CA but not in Ishikawa cells. Induction of apoptosis in AN3CA cells was not mediated by the p53 pathway activation but was rather associated to reduced AKT phosphorylation. Interestingly, immunofluorescence analysis evidenced that endometrioid adenocarcinoma tissues displayed, similarly to AN3CA cells, also a GRP78 plasma membrane localization. These data suggest that GRP78 and its plasma membrane localization, might play a role in endometrial cancer development and progression and might constitute a novel target for the treatment of endometrial cancer. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 13, 2014   doi: 10.1002/jcp.24578   open full text
• Reelin/DAB‐1 Signaling in the Embryonic Limb Regulates the Chondrogenic Differentiation of Digit Mesodermal Progenitors.
  Manuel J. Diaz‐Mendoza, Carlos I. Lorda‐Diez, Juan A. Montero, Juan A. Garcia‐Porrero, Juan M. Hurle.
  Journal of Cellular Physiology. February 12, 2014
  Reelin is a bioactive component of some extracellular matrices. Most studies on this signaling glycoprotein have been performed in the developing nervous system, where Reelin binds to the very‐low‐density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2) of target cells. This induces phosphorylation of the intracellular adaptor protein Disabled‐1 (Dab‐1), which subsequently activates downstream effectors to regulate important aspects of neuroblast biology. Here, we show that the components of the Reelin signaling pathway exhibit a dynamic expression pattern during the development of the digits in chick and mouse embryonic limbs. Reelin and Dab‐1 are highly expressed in the differentiating digit cartilages and tendinous blastemas. Immunolabeling of phospho‐Dab‐1 indicates that the pattern of gene expression correlates with zones of active signaling. Intense signaling is also present in the early stages of cartilage differentiation in micromass cultures of digit mesodermal progenitors. In this in vitro assay, disruption of the Reelin signaling pathway by gene silencing causes cystoskeletal and cell shape modifications accompanied by reduced chondrogenesis and down‐regulation of specific cartilage molecular markers. Of note, Scleraxis and Six2, which are master genes of tendinous blastemas, become upregulated in these experiments. We further show that the receptors ApoER2 and VLDLR are differentially expressed in cartilage and tendons and that these receptors show temporal expression differences in the micromass cultures. Sox9 and other chondrogenic markers were downregulated in micromass cultures after ApoER2 gene silencing, while gene silencing of VLDLR up‐regulates Scleraxis. In summary our findings provide evidence of a role for Reelin signaling in skeletogenesis that promotes chondrogenesis through ApoER2 and inhibits tenogenic differentiation through VLDLR. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 12, 2014   doi: 10.1002/jcp.24576   open full text
• Altered Histone Mark Deposition and DNA Methylation at Homeobox Genes in Human Oral Squamous Cell Carcinoma Cells.
  Katarzyna M Marcinkiewicz, Lorraine J Gudas.
  Journal of Cellular Physiology. February 12, 2014
  We recently reported a role of Polycomb repressive complex 2 (PRC2) and PRC2 trimethylation of histone 3 lysine 27 (H3K27me3) in the regulation of homeobox (HOX) (Marcinkiewicz and Gudas, 2013) gene transcript levels in human oral keratinocytes (OKF6‐TERT1R) and tongue squamous cell carcinoma (SCC) cells. Here, we assessed both the levels of various histone modifications at a subset of homeobox genes and genome wide DNA methylation patterns in OKF6‐TERT1R and SCC‐9 cells by using ERRBS (enhanced reduced representation bisulfite sequencing). We detected the H3K9me3 mark at HOXB7, HOXC10, HOXC13 and HOXD8 at levels higher in OKF6‐TERT1R than in SCC‐9 cells; at IRX1 and SIX2 the H3K9me3 levels were conversely higher in SCC‐9 than in OKF6‐TERT1R. The H3K79me3 mark was detectable only at IRX1 in OKF6‐TERT1R and at IRX4 in SCC‐9 cells. The levels of H3K4me3 and H3K36me3 marks correlate with the transcript levels of the assessed homeobox genes in both OKF6‐TERT1R and SCC‐9. We detected generally lower CpG methylation levels on DNA in SCC‐9 cells at annotated genomic regions which were differentially methylated between OKF6‐TERT1R and SCC‐9 cells; however, some genomic regions, including the HOX gene clusters, showed DNA methylation at higher levels in SCC‐9 than OKF6‐TERT1R. Thus, both altered histone modification patterns and changes in DNA methylation are associated with dysregulation of homeobox gene expression in human oral cavity SCC cells, and this dysregulation potentially plays a role in the neoplastic phenotype of oral keratinocytes. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 12, 2014   doi: 10.1002/jcp.24577   open full text
• Non‐Cell Autonomous or Secretory Tumor Suppression.
  Christelle En Lin Chua, Shu Ning Chan, Bor Luen Tang.
  Journal of Cellular Physiology. February 06, 2014
  Many malignancies result from deletions or loss‐of‐function mutations in one or more tumor suppressor genes, the products of which curb unrestrained growth or induce cell death in those with dysregulated proliferative capacities. Most tumor suppressors act in a cell autonomous manner, and only very few proteins are shown to exert a non‐cell autonomous tumor suppressor function on other cells. Examples of these include members of the secreted frizzled‐related protein (SFRP) family and the secreted protein acidic and rich in cysteine (SPARC)‐related proteins. Very recent findings have, however, considerably expanded our appreciation of non‐cell autonomous tumor suppressor functions. Broadly, this may occur in two ways. Intracellular tumor suppressor proteins within cells could in principle inhibit aberrant growth of neighboring cells by conditioning an antitumor microenvironment through secreted factors. This is demonstrated by an apparent non‐cell autonomous tumor suppressing property of p53. On the other hand, a tumor suppressor produced by a cell may be secreted extracellularly, and taken up by another cell with its activity intact. Intriguingly, this has been recently shown to occur for the phosphatase and tensin homolog (PTEN) by both conventional and unconventional modes of secretion. These recent findings would aid the development of therapeutic strategies that seek to reinstate tumor suppression activity in therapeutically recalcitrant tumor cells, which have lost it in the first place. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  February 06, 2014   doi: 10.1002/jcp.24574   open full text
• Calcineurin Regulates Progressive Motility Activation of Rhinella (Bufo) arenarum Sperm Through Dephosphorylation of PKC Substrates.
  Dario Krapf, Emma ÓBrien, María P. Maidagán, Enrique O. Morales, Pablo E. Visconti, Silvia E. Arranz.
  Journal of Cellular Physiology. January 31, 2014
  Animals with external fertilization, as amphibians, store their sperm in a quiescent state in the testis. When spermatozoa are released into natural fertilization media, the hypotonic shock triggers activation of sperm motility. Rhinella (Bufo) arenarum sperm are immotile in artificial seminal plasma (ASP, resembling testicular plasma tonicity) but acquire in situ flagellar beating upon dilution. However, if components from the egg shelly coat are added to this medium, motility shifts to a progressive pattern. Recently, we have shown that the signal transduction pathway required for in situ motility activation involves a rise in intracellular cAMP through a transmembrane adenylyl cyclase and activation of PKA, mostly in the midpiece and in the sperm head. In this report, we demonstrate that activation of calcineurin (aka PP2B and PPP3) is required for the shift from in situ to progressive sperm motility. The effect of calcineurin is manifested by dephosphorylation of PKC substrates, and can be promoted by intracellular calcium rise by Ca2+ ionophore. Both phosphorylated PKC substrates and calcineurin localized to the flagella, indicating a clear differentiation between compartmentalization of PKA and calcineurin pathways. Moreover, no crosstalk is observed between these signaling events, even though both pathways are required for progressive motility acquisition as discussed. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  January 31, 2014   doi: 10.1002/jcp.24571   open full text
• CD40 Signaling Drives B Lymphocytes Into an Intermediate Memory‐Like State, Poised Between Naïve and Plasma Cells.
  Mala Upadhyay, G.Krishna Priya, P. Ramesh, M.B. Madhavi, Satyajit Rath, Vineeta Bal, Anna George, Tushar Vaidya.
  Journal of Cellular Physiology. January 31, 2014
  Immunological memory comprising of antigen‐specific B and T cells contributes to the acquisition of long‐term resistance to pathogens. Interactions between CD40 on B cells and CD40L on T cells are responsible for several aspects of acquired immune responses including generation of memory B cells. In order to gain insights into events leading to memory B cell formation, we analyzed the genome‐wide expression profile of murine naive B cells stimulated in the presence of anti‐CD40. We have identified over 8000 genes whose expression is altered minimally 1.5 fold at least at one time point over a three day time course. The array analysis indicates that changes in expression level of maximum number of these genes occur within 24 hours of anti‐CD40 treatment. In parallel, we have studied the events following CD40 ligation by examining the expression of known regulators of naive B cell to plasma cell transition, including Pax5 and BLIMP1. The expression profile of these regulatory genes indicates firstly, that CD40 signaling activates naïve B cells to a phenotype that is intermediate between the naive and plasma cell stages of the B cell differentiation. Secondly, the major known regulator of plasma cell differentiation, BLIMP1, gets irreversibly down regulated upon anti‐CD40 treatment. Additionally, our data reveals that CD40 signaling mediated BLIMP1 down regulation occurs by non‐Pax5/non‐Bcl6 dependent mechanisms, indicating novel mechanisms at work that add to the complexity of understanding of B cell master regulatory molecules like BLIMP1 and Pax5. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  January 31, 2014   doi: 10.1002/jcp.24572   open full text
• Midbrain Cues Dictate Differentiation of Human Dental Pulp Stem Cells Towards Functional Dopaminergic Neurons.
  Mohammad Kanafi, Debanjana Majumdar, Ramesh Bhonde, Indrani Datta.
  Journal of Cellular Physiology. January 29, 2014
  Dental pulp originating from the neural crest is considered a better source of postnatal stem cells for cell‐based therapies in neurodegenerative diseases. Dental Pulp Stem Cells (DPSCs) have been shown to differentiate into cell‐types of cranial neural crest ontology; however, their ability to differentiate to functional neurons of the central nervous system remains to be studied. We hypothesized that midbrain cues might commit DPSCs to differentiate to functional dopaminergic cell‐type. As expected, DPSCs in their naïve state spontaneously expressed early and mature neuronal markers like nestin, musashi12, β tubulin III and Map2ab. On exposure to midbrain cues (sonic hedgehog, fibroblast growth factor 8 and basic fibroblast growth factor), DPSCs showed upregulation of dopaminergic neuron‐specific transcription factors Nuclear Receptor related protein 1 (Nurr1), Engrailed 1 (En1) and paired‐like homeodomain transcription factor 3 (Pitx3) as revealed by real‐time RT‐PCR. Immunofluorescence and flow cytometry analysis showed enhanced expression of mature neuronal marker Map2ab and dopaminergic‐neuronal markers [tyrosine hydroxylase (TH), En1, Nurr1 and Pitx3], with nearly 77% of the induced DPSCs positive for TH. Functional studies indicated that the induced DPSCs could secrete dopamine constitutively and upon stimulation with potassium chloride (KCl) and adenosine triphosphate (ATP), as measured by dopamine ELISA. Additionally, the induced DPSCs showed intracellular Ca2+ influx in the presence of KCl, unlike control DPSCs. ATP‐stimulated Ca2+ influx was observed in control and induced DPSCs, but only the induced cells secreted dopamine. Our data clearly demonstrates for the first time that DPSCs in the presence of embryonic midbrain cues show efficient propensity towards functional dopaminergic cell‐type. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  January 29, 2014   doi: 10.1002/jcp.24570   open full text
• Phosphorylation‐Regulated Degradation of the Tumor‐Suppressor Form of PED by Chaperone‐Mediated Autophagy in Lung Cancer Cells.
  Cristina Quintavaller, Stefaniadi Costanzo, Ciro Zanca, Immaculada Tasset, Alessandro Fraldi, Mariarosaria Incoronato, Peppino Mirabelli, Maria Monti, Andrea Ballabio, Piero Pucci, Ana Maria Cuervo, Aerolama Condorelli.
  Journal of Cellular Physiology. January 29, 2014
  PED/PEA‐15 is a death effector domain (DED) family member with a variety of effects on cell growth and metabolism. To get further insight into the role of PED in cancer, we aimed to find new PED interactors. Using tandem affinity purification, we identified HSC70 (Heat Shock Cognate Protein of 70 kDa)—which, among other processes, is involved in chaperone‐mediated autophagy (CMA)—as a PED‐interacting protein. We found that PED has two CMA‐like motifs (i.e., KFERQ), one of which is located within a phosphorylation site, and demonstrate that PED is a bona fide CMA substrate and the first example in which phosphorylation modifies the ability of HSC70 to access KFERQ‐like motifs and target the protein for lysosomal degradation. Phosphorylation of PED switches its function from tumor suppression to tumor promotion, and we show that HSC70 preferentially targets the unphosphorylated form of PED to CMA. Therefore, we propose that the up‐regulated CMA activity characteristic of most types of cancer cell enhances oncogenesis by shifting the balance of PED function toward tumor promotion. This mechanism is consistent with the notion of a therapeutic potential for targeting CMA in cancer, as inhibition of this autophagic pathway may help restore a physiological ratio of PED forms. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  January 29, 2014   doi: 10.1002/jcp.24569   open full text
• Wnt5a Suppresses Colon Cancer by Inhibiting Cell Proliferation and Epithelial–Mesenchymal Transition.
  Runfen Cheng, Baocun Sun, Zhiyong Liu, Xiulan Zhao, Lisha Qi, Yixian Li, Qiang Gu.
  Journal of Cellular Physiology. January 24, 2014
  Colon cancer remains one of the lethal malignancies in the world. Aberrant activation of canonical Wnt/β‐catenin signaling pathway has been observed in colon cancer. In contrast, the non‐canonical Wnt signaling functions remain obscure. Wnt5a is a representative non‐canonical Wnt ligand which has gained extensive attention nowadays. Wnt5a has been shown to play an important role in EMT in prostate cancer and melanoma, but its role in colon cancer is still ambiguous. Here we have evaluated Wnt5a expression in a large cohort of 217 colon cancers by immunohistochemistry and analyzed its correlation with clinicopathologic characteristics. We found that expression of Wnt5a was diminished significantly in majority of primary colon cancers and negatively related with EMT biomarkers. To further enlighten the mechanism which Wnt5a regulates EMT in vitro, we established ectopic Wnt5a expression models. Protein analysis demonstrated that Wnt5a inhibited EMT and antagonized canonical Wnt signaling in colon cancer cells. Overexpression of Wnt5a impaired cell motility and invasion and inhibited cell proliferation by manipulating Bax. Moreover, Wnt5a suppressed the tumor growth in nude mice and impaired tumorigenicity in vivo. Wnt5a also induced intracellular calcium and activated non‐canonical Wnt/Ca2+ signaling in colon cancer. In summary, although Wnt5a was down‐regulated in majority of colon cancers, enhanced Wnt5a expression predict preferable outcome in colon cancer patients. Our findings indicate that Wnt5a might act as tumor suppressor by inhibiting cell proliferation and attenuating EMT in colon cancer cells. Wnt5a could be used as a novel prognostic marker and/or therapeutic target for colon cancer in the future. J. Cell. Physiol. © 2014 Wiley Periodicals, Inc.

  January 24, 2014   doi: 10.1002/jcp.24566   open full text
• In Vivo Deletion of CAR Resulted in High Bone Mass Phenotypes in Male Mice.
  Hwa Young Cho, Ju‐Yeon Jung, Hyojung Park, Jae‐Yeon Yang, Solip Jung, Jee Hyun An, Sun Wook Cho, Sang Wan Kim, Seong Yeon Kim, Jung Eun Kim, Young Joo Park, Chan Soo Shin.
  Journal of Cellular Physiology. January 21, 2014
  Constitutive androstane receptor (CAR) was originally identified as xenobiotic sensor that regulates the expression of cytochrome P450 genes. However, recent studies suggest that this nuclear receptor is also involved in the regulation of energy metabolism including glucose and lipid homeostasis. This study investigated the role of CAR in the regulation of bone mass in vivo using CAR−/− mice. Endogenous mRNA expression of CAR was observed in both primary osteoblasts and osteoclast precursors. CAR−/− mice have exhibited significant increase in whole body bone mineral density (BMD) by 9.5% (P < 0.01) and 5.5% (P < 0.05) at 10 and 15 weeks of age, respectively, compared with WT mice in males. Microcomputed tomography analysis of proximal tibia demonstrated a significant increase in trabecular bone volume (62.7%), trabecular number (54.1%) in male CAR−/− mice compared with WT mice. However, primary culture of calvarial cells exhibited no significant changes in osteogenic differentiation potential between CAR−/− and WT. In addition, the number of tartrate‐resistant acid‐phosphatase positive osteoclasts in the femur and serum level of CTx was not different between CAR−/− and WT mice. The higher BMD and microstructural parameters were not observed in female mice. Interestingly, serum level of testosterone in male CAR−/− mice was 2.5‐fold higher compared with WT mice and the mRNA expressions of Cyp2b9 and 2b10 in the liver, which regulate testosterone metabolism, were significantly down‐regulated in male CAR−/− mice. Furthermore, the difference in BMD between CAR−/− and WT mice disappeared at 8 weeks after performing orchiectomy. CAR−/− mice also exhibited significant increase in serum 1,25(OH)2D3 levels but Cyp 27B1 which converts 25(OH)D3 to 1,25(OH)2D3 was significantly down‐regulated compared to WT mice. These results suggest that in vivo deletion of CAR resulted in higher bone mass, which appears to be a result from reduced metabolism of testosterone due to down‐regulation of Cyp2b. J. Cell. Physiol. 229: 561–571, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24478   open full text
• Hypoxia Simultaneously Alters Satellite Cell‐Mediated Angiogenesis and Hepatocyte Growth Factor Expression.
  K.L. Flann, C.R. Rathbone, L.C. Cole, X. Liu, R.E. Allen, R.P. Rhoads.
  Journal of Cellular Physiology. January 21, 2014
  Skeletal muscle regeneration is a multifaceted process requiring the spatial and temporal coordination of myogenesis as well as angiogenesis. Hepatocyte growth factor (HGF) plays a pivotal role in myogenesis by activating satellite cells (SC) in regenerating muscle and likely plays a role as a contributor to revascularization. Moreover, repair of a functional blood supply is critical to ameliorate tissue ischemia and restore skeletal muscle function, however effects of hypoxia on satellite cell‐mediated angiogenesis remain unclear. The objective of this study was to examine the role of HGF and effect of hypoxia on the capacity of satellite cells to promote angiogenesis. To characterize the role of HGF, a microvascular fragment (MVF) culture model coupled with satellite cell conditioned media (CM) was employed. The activity of HGF was specifically blocked in SC CM reducing sprout length compared to control CM. In contrast, MVF sprout number did not differ between control or HGF‐deficient SC CM media. Next, we cultured MVF in the presence of CM from satellite cells exposed to normoxic (20% O2) or hypoxic (1% O2) conditions. Hypoxic CM recapitulated a MVF angiogenic response identical to HGF deficient satellite cell CM. Hypoxic conditions increased satellite cell HIF‐1α protein abundance and VEGF mRNA abundance but decreased HGF mRNA abundance compared to normoxic satellite cells. Consistent with reduced HGF gene expression, HGF promoter activity decreased during hypoxia. Taken together, this data indicates that hypoxic modulation of satellite cell‐mediated angiogenesis involves a reduction in satellite cell HGF expression. J. Cell. Physiol. 229: 572–579, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24479   open full text
• Amsacrine suppresses matrix metalloproteinase‐2 (MMP‐2)/MMP‐9 expression in human leukemia cells.
  Wen‐Hsin Liu, Ying‐Jung Chen, Jen‐Hung Chien, Long‐Sen Chang.
  Journal of Cellular Physiology. January 21, 2014
  This study explores the suppression mechanism of amsacrine (4‐(9‐Acridinylamino)‐N‐(methanesulfonyl)‐m‐anisidine hydrochloride) on matrix metalloproteinase‐2 (MMP‐2) and MMP‐9 expression in human leukemia cells. Amsacrine attenuated cell invasion with decreased MMP‐2/MMP‐9 protein expression and mRNA levels in U937, Jurkat, HL‐60, K562, KU812, and MEG‐01 cells. Moreover, amsacrine reduced both MMP‐2/MMP‐9 promoter luciferase activity and MMP‐2/MMP‐9 mRNA stability in leukemia cells. Studies on amsacrine‐treated U937 cells revealed that amsacrine‐elicited ROS generation induced JNK and p38 MAPK activation but reduced the phospho‐ERK level. Amsacrine‐induced ERK inactivation and p38 MAPK/JNK activation were demonstrated to suppress MMP‐2/MMP‐9 promoter luciferase activity and promote MMP‐2/MMP‐9 mRNA decay, respectively. p38 MAPK/JNK activation led to up‐regulation of protein phosphatase 2A catalytic subunit α (PP2Acα) in amsacrine‐treated U937 cells. Okadaic acid (PP2A inhibitor) treatment increased MMP‐2/MMP‐9 mRNA stability in amsacrine‐treated cells, whereas PP2Acα over‐expression increased MMP‐2/MMP‐9 mRNA decay. Amsacrine‐induced MMP‐2/MMP‐9 down‐regulation was also related to PP2Acα up‐regulation on Jurkat, HL‐60, K562, KU812, and MEG‐01 cells. Collectively, our data indicate that amsacrine induces MMP‐2/MMP‐9 down‐regulation via simultaneous suppression of genetic transcription and mRNA stability in human leukemia cells. J. Cell. Physiol. 229: 588–598, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24481   open full text
• Fn14 Promotes Differentiation of Human Mesenchymal Stem Cells into Heart Valvular Interstitial Cells by Phenotypic Characterization.
  Wei Huang, Ding‐Zhang Xiao, Yigang Wang, Zhi‐Xin Shan, Xiao‐Ying Liu, Qiu‐Xiong Lin, Min Yang, Jian Zhuang, Yangxin Li, Xi‐Yong Yu.
  Journal of Cellular Physiology. January 21, 2014
  Despite the fact that tissue engineered heart valves (TEHV) hold great promise for heart valve disease treatment, one of the challenges is to find suitable seeding cells. Bone marrow derived mesenchymal stem cells (MSCs) were considered to be one of the best seed cell sources. In this study we propose a novel approach to promote stem cell differentiation into the seed cells of TEHV, valvular interstitial cells (VICs). Newly induced MSCs (iMSCs) were created from a co‐culture niche in which healthy human donor derived MSCs were co‐cultured with cardiac fibroblasts (H9C2 cell line). Then iMSCs were transfected with either a mock vector (iMSCsmock) as controls or with a vector that overexpresses thefibroblast inducible factor 14 (Fn14) gene (iMSCsFn14). Immunofluorescence staining was performed to assay VIC differentiation. Western blot analysis was performed to analyze the involved signaling pathway. The results demonstrate that the expression of α‐smooth muscle actin (SMA) was significantly higher in iMSCsFn14 as compared with iMSCmock, and MSC, and also had higher co‐alignment of α‐actinin and stress fiber (F‐actin) in bundles. Additionally, increased biosynthesis of extracellular matrix (ECM) proteins including collagen I, collagen III, and fibronection were observed in iMSCsFn14 in comparison with iMSCsmock. These data observed in iMSCsFn14 were in accordance with VIC phenotype from normal heart valves. In addition, the PI3K/Akt signaling pathway was activated in iMSCsFn14 which allowed higher Akt phosphorylation (p‐Akt) levels and SMA levels, whereas, it was attenuated by LY294002 (PI3K/Akt inhibitor). These new findings of the effect of Fn14 on VIC‐like cell differentiation may provide a novel therapeutic strategy for heart valve disease treatment. J. Cell. Physiol. 229: 580–587, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24480   open full text
• RNA Polymerase II CTD Modifications: How Many Tales From a Single Tail.
  Giuliana Napolitano, Luigi Lania, Barbara Majello.
  Journal of Cellular Physiology. January 21, 2014
  Eukaryote's RNA polymerases II (RNAPII) have the feature to contain, at the carbossi‐terminal region of their largest subunit Rpb1, a unique CTD domain. Rpb1‐CTD is composed of an increasing number of repetitions of the Y1S2P3T4S5P6S7 heptad that goes in parallel with the developmental level of organisms. Because of its composition, the CTD domain has a huge structural plasticity; virtually all the residues can be subjected to post‐translational modifications and the two prolines can either be in cis or trans conformations. In light of these features, it is reasonable to think that different specific nuances of CTD modification and interacting factors take place not only on different gene promoters but also during different stages of the transcription cycle and reasonably might have a role even if the polymerase is on or off the DNA template. Rpb1‐CTD domain is involved not only in regulating transcriptional rates, but also in all co‐transcriptional processes, such as pre‐mRNA processing, splicing, cleavage, and export. Moreover, recent studies highlight a role of CTD in DNA replication and in maintenance of genomic stability and specific CTD‐modifications have been related to different CTD functions. In this paper, we examine results from the most recent CTD‐related literature and give an overview of the general function of Rpb1‐CTD in transcription, transcription‐related and non transcription‐related processes in which it has been recently shown to be involved in. J. Cell. Physiol. 229: 538–544, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24483   open full text
• Endocan Elicits Severe Vascular Inflammatory Responses In Vitro and In Vivo.
  Wonhwa Lee, Sae‐Kwang Ku, Shin‐Woo Kim, Jong‐Sup Bae.
  Journal of Cellular Physiology. January 21, 2014
  Endocan is a proteoglycan secreted by endothelial cells under the control of inflammatory cytokines. The aim of this study was to evaluate the effects of endocan on proinflammatory responses and on septic mice and underlying mechanisms. Human umbilical vein endothelial cells (HUVECs) or mice were exposed to lipopolysaccharide (LPS) or endocan with or without neutralizing endocan antibody. Mice were subjected to cecal ligation and puncture (CLP) surgery with or without neutralizing endocan antibody. Endocan was highly released by LPS and it enhanced proinflammatory responses. In a CLP‐induced sepsis model, endocan was also highly released, but this release was prevented by administration of neutralizing endocan antibody. Circulating levels of endocan measured in patients admitted to the intensive care unit with sepsis were significantly elevated compared with control donors. Furthermore, the administration of endocan antibody reduced CLP‐induced sepsis mortality. This study shows endocan can elicit severe inflammatory responses and inhibiting endocan release offers a potential strategy for treating sepsis. J. Cell. Physiol. 229: 620–630, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24485   open full text
• Mathematical Model of Liver Regeneration in Human Live Donors.
  V. Periwal, J.R. Gaillard, L. Needleman, C. Doria.
  Journal of Cellular Physiology. January 21, 2014
  Liver regeneration after injury occurs in many mammals. Rat liver regenerates after partial hepatectomy over a period of 2 weeks while human liver regeneration takes several months. Notwithstanding this enormous difference in time‐scales, with new data from five human live liver transplant donors, we show that a mathematical model of rat liver regeneration can be transferred to human, with all biochemical interactions and signaling unchanged. Only six phenomenological parameters need change, and three of these parameter changes are rescalings of rate constants by the ratio of human lifespan to rat lifespan. Data from three donor subjects with approximately equal resections were used to fit the three parameters and the data from the other two donor subjects was used to independently verify the fit. J. Cell. Physiol. 229: 599–606, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24482   open full text
• Microvesicle‐Shuttled miR‐130b Reduces Fat Deposition in Recipient Primary Cultured Porcine Adipocytes by Inhibiting PPAR‐γ Expression.
  Shifeng Pan, Xiaojing Yang, Yimin Jia, Runsheng Li, Ruqian Zhao.
  Journal of Cellular Physiology. January 21, 2014
  Obesity is a worldwide epidemic, and a risk factor for cardiovascular disease and type 2 diabetes. Consequently, the development of safe and effective anti‐obesity drugs is an area of ongoing clinical interest. MicroRNAs play a vital role in anti‐obesity by inhibiting the expression of genes involved in adipogenesis and lipogenesis. However, the clinical application of miRNAs has been limited by a lack of appropriate delivery systems. The discovery of microvesicles (MVs) has shed new light on the search for more efficient drug transport tools. In a previous study, we demonstrated that miRNA‐130b suppressed fat deposition by inhibiting PPAR‐γ expression. In order to demonstrate whether miRNA‐130b can be packaged into MVs and function as an endogenous form of miRNA‐130b in recipient cells, we transfected HeLa‐229 cells with plasmid to overexpress miRNA‐130b. This enabled HeLa‐229 cells to selectively package miRNA‐130b into MVs and actively secrete the miRNA‐130b enriched MVs into the culture media. We further verified that MVs enriched with miRNA‐130b contain elevated concentrations of Argonaute 2 and heat shock protein 90α which are known to protect the circulating miRNAs from degradation. Exposure of primary cultured porcine adipocytes to purified, miRNA‐130b‐enriched MVs resulted in a significant down‐regulation of PPAR‐γ expression which was associated with reduced adipogenesis and lipogenesis. Taken together, our results suggest that MVs may provide an effective transport systems for the deliver of miRNAs for therapeutic use. We also showed that MV‐shuttled miRNA‐130b inhibited adipogenesis and lipogenesis, and reduced fat deposition in recipient adipocytes by targeting PPAR‐γ. J. Cell. Physiol. 229: 631–639, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24486   open full text
• Smad6 Suppresses the Growth and Self‐Renewal of Hepatic Progenitor Cells.
  Ze‐Yang Ding, Hui‐Fang Liang, Guan‐Nan Jin, Wei‐Xun Chen, Wei Wang, Pran K. Datta, Ming‐Zhi Zhang, Bixiang Zhang, Xiao‐Ping Chen.
  Journal of Cellular Physiology. January 21, 2014
  Activation of hepatic progenitor cells (HPCs) is commonly observed in chronic liver disease and Wnt/β‐catenin signaling plays a crucial role in the expansion of HPCs. However, the molecular mechanisms that regulate the activation of Wnt/β‐catenin signaling in the liver, especially in HPCs, remain largely elusive. Here, we reported that ectopic expression of Smad6 suppressed the proliferation and self‐renewal of WB‐F344 cells, a HPC cell line. Mechanistically, we found that Smad6 inhibited Wnt/β‐catenin signaling through promoting the interaction of C‐terminal binding protein (CtBP) with β‐catenin/T‐cell factor (TCF) complex to inhibit β‐catenin mediated transcriptional activation in WB‐F344 cells. We used siRNA targeting β‐catenin to demonstrate that Wnt/β‐catenin signaling was required for the proliferation and self‐renewal of HPCs. Taken together, these results suggest that Smad6 is a regulatory molecule which regulates the proliferation, self‐renewal and Wnt/β‐catenin signaling in HPCs. J. Cell. Physiol. 229: 651–660, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24488   open full text
• Adaptive Responses of Mitochondria to Mild Copper Deprivation Involve Changes in Morphology, OXPHOS Remodeling and Bioenergetics.
  Lina María Ruiz, Erik L. Jensen, Rodrigo I. Bustos, Graciela Argüelloa, Ricardo Gutierrez‐Garcia, Mauricio González, Claudia Hernández, Rodolfo Paredes, Felipe Simon, Claudia Riedel, David Ferrick, Alvaro A. Elorza.
  Journal of Cellular Physiology. January 21, 2014
  Copper is an essential cofactor of complex IV of the electron transfer chain, and it is directly involved in the generation of mitochondrial membrane potential. Its deficiency induces the formation of ROS, large mitochondria and anemia. Thus, there is a connection between copper metabolism and bioenergetics, mitochondrial dynamics and erythropoiesis. Copper depletion might end in cellular apoptosis or necrosis. However, before entering into those irreversible processes, mitochondria may execute a series of adaptive responses. Mitochondrial adaptive responses (MAR) may involve multiple and diverse mechanisms for preserving cell life, such as mitochondrial dynamics, OXPHOS remodeling and bioenergetics output. In this study, a mild copper deficiency was produced in an animal model through intraperitoneal injections of bathocuproine disulfonate in order to study the MAR. Under these conditions, a new type of mitochondrial morphology was discovered in the liver. Termed the “butternut squash” mitochondria, it coexisted with normal and swollen mitochondria. Western blot analyses of mitochondrial dynamics proteins showed an up‐regulation of MFN‐2 and OPA1 fusion proteins. Furthermore, isolated liver mitochondria displayed OXPHOS remodeling through a decrease in supercomplex activity with a concomitant increase at an individual level of complexes I and IV, higher respiratory rates at complex I and II levels, higher oligomycin‐insensitive respiration, and lower respiratory control ratio values when compared to the control group. As expected, total ATP and ATP/ADP values were not significantly different, since animal's health was not compromised. As a whole, these results describe a compensatory and adaptive response of metabolism and bioenergetics under copper deprivation. J. Cell. Physiol. 229: 607–619, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24484   open full text
• TNF‐α Upregulates Sclerostin Expression in Obese Mice Fed a High‐Fat Diet.
  Kyunghwa Baek, Hyo Rin Hwang, Hyun‐Jung Park, Arang Kwon, Abdul S. Qadir, Seong‐Hee Ko, Kyung Mi Woo, Hyun‐Mo Ryoo, Gwan‐Shik Kim, Jeong‐Hwa Baek.
  Journal of Cellular Physiology. January 21, 2014
  Sclerostin decreases bone mass by antagonizing the Wnt signaling pathway. We examined whether obesity‐induced bone loss is associated with the expression of sclerostin. Five‐week‐old male mice were assigned to one of two groups (n = 10 each) and fed either a control diet (10% kcal from fat; CON) or a high‐fat diet (60% kcal from fat; HF) for 12 weeks. Thex final body weight and whole body fat mass of the HF mice were higher than those of the CON mice. The distal femur cancellous bone mineral density and bone formation rate was lower in HF mice than in CON mice. The percent erosion surface was higher in the HF mice than the CON mice. The serum levels and femoral osteocytic protein expression levels of tumor necrosis factor‐α (TNF‐α) were significantly higher in HF mice than in CON mice. Sclerostin mRNA levels and osteocytic sclerostin protein levels in femoral cortex were also higher in HF mice than in CON mice. Sclerostin expression in MLO‐Y4 osteocytes increased with TNF‐α treatment, and TNF‐α‐induced sclerostin expression was blocked by the inhibition of NF‐κB activation. Chromatin immunoprecipitation and a luciferase reporter assay demonstrated that NF‐κB directly binds to the NF‐κB binding elements on the mouse sost promoter and stimulates sclerostin expression. These results support a model in which, in the context of obesity or other inflammatory diseases that increase the production of TNF‐α, TNF‐α upregulates the expression of sclerostin through NF‐κB signaling pathway, thus contributing to bone loss. J. Cell. Physiol. 229: 640–650, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24487   open full text
• Impact of S100A8/A9 Expression on Prostate Cancer Progression In Vitro and In Vivo.
  Sina Grebhardt, Karin Müller‐Decker, Felix Bestvater, Michal Hershfinkel, Doris Mayer.
  Journal of Cellular Physiology. January 21, 2014
  The proinflammatory S100A8/A9 proteins, which are expressed in myeloid cells under physiological conditions, are strongly expressed in human prostate cancer epithelial cells. Their role in the tumor cells and in tumor progression is largely unclear. We established a prostate cancer epithelial cell line (PC‐3 TO‐A8/A9) expressing S100A8 and S100A9 simultaneously under doxycycline control, to study the role of S100A8/A9 on tumor growth and infiltration of immune cells in subcutaneous xenografts in male NMRI nu/nu mice. Colonization of distant organs was studied after intracardial injection of the tumor cells in male NOD/SCID mice. PC‐3 TO‐A8/A9 cells grown in vitro and subcutaneous xenografts in mice not treated with doxycycline expressed high levels of S100A8/A9 mRNA and protein, whereas doxycycline treatment suppressed S100A8/A9 expression. S100A8/A9 expression did not significantly alter growth rate and invasion of the subcutaneous tumors into surrounding tissues. However, S100A8/A9 expression caused increased infiltration of immune cells, especially neutrophils. In intracardially injected mice sporadic tumor settlement was observed in muscle and lymph nodes. Colonies of tumor cells and micro‐metastases were observed in the lung of 64.3% (9 out of 14) of mice not treated with doxycycline and in 33.3% (5 out of 15) of mice treated with doxycycline. Our data demonstrate for the first time that S100A8/A9 expression in epithelial cancer cells causes enhanced infiltration of immune cells, especially neutrophils, and stimulates settlement of the cancer cells in the lung. J. Cell. Physiol. 229: 661–671, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24489   open full text
• Connective Tissue Growth Factor (CTGF/CCN2) Negatively Regulates BMP‐2 Induced Osteoblast Differentiation and Signaling.
  Christina Mundy, Maureen Gannon, Steven N. Popoff.
  Journal of Cellular Physiology. January 21, 2014
  Connective tissue growth factor (CTGF/CCN2) and bone morphogenetic protein (BMP)‐2 are both produced and secreted by osteoblasts. Both proteins have been shown to have independent effects in regulating osteoblast proliferation, maturation and mineralization. However, how these two proteins interact during osteoblast differentiation remains unknown. In this study, we utilized two cell culture model systems, osteoblasts derived from CTGF knockout (KO) mice and osteoblasts infected with an adenovirus which over‐expresses CTGF (Ad‐CTGF), to investigate the effects of CTGF and BMP‐2 on osteoblast development and function in vitro. Contrary to a previously published report, osteoblast maturation and mineralization were similar in osteogenic cultures derived from KO and WT calvaria in the absence of BMP‐2 stimulation. Interestingly, in KO and WT osteoblast cultures stimulated with BMP‐2, the KO osteoblasts exhibited enhanced osteoblast differentiation. This increase in osteoblast differentiation was accompanied by increased protein levels of phosphorylated Smad 1/5/8 and mRNA expression levels of bone morphogenetic protein receptor Ib. We also examined osteoblast differentiation in cultures that were infected with an adenoviral‐CTGF vector (Ad‐CTGF) and in controls. Continuous over‐expression of CTGF resulted in decreased osteoblast maturation and mineralization in both unstimulated and BMP‐2 stimulated cultures. Impaired osteoblast differentiation in cultures over‐expressing CTGF was accompanied by decreased protein levels of phosphorylated Smad 1/5/8. Collectively, the data from these studies demonstrate that CTGF acts to negatively regulate BMP‐2 induced signaling and osteoblast differentiation, and warrant additional studies to determine the precise mechanism(s) responsible for this effect. J. Cell. Physiol. 229: 672–681, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24491   open full text
• The Pivotal Role of microRNA‐155 in the Control of Cancer.
  Zhaolin Chen, Taotao Ma, Cheng Huang, Tingting Hu, Jun Li.
  Journal of Cellular Physiology. January 21, 2014
  microRNAs (miRNAs) are emerging as important gene expression regulators linked to various biological processes at a posttranscriptional level. miRNAs have been known to play important roles in cell proliferation, cell differentiation, and apoptosis. Recently, accumulate studies indicate that up‐regulation of miR‐155 has been described in several types of human tumors. miR‐155 has been considered to act as an oncogene or a tumor suppressor, depending on tumor system. Silencing oncomiRs or gene therapy approaches could be an effective therapeutic approach against tumor. Here we review the current knowledge on the functional role of miR‐155 in the control of various cancers. J. Cell. Physiol. 229: 545–550, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24492   open full text
• Transcription Factor Sp1 Regulates T‐Type Ca2+ Channel CaV3.1 Gene Expression.
  Ricardo González‐Ramírez, Elizabeth Martínez‐Hernández, Alejandro Sandoval, Ricardo Felix.
  Journal of Cellular Physiology. January 21, 2014
  Voltage‐gated T‐type Ca2+ (CaV3) channels mediate a number of physiological events in developing and mature cells, and are implicated in neurological and cardiovascular diseases. In mammals, there are three distinct T‐channel genes (CACNA1G, CACNA1H, and CACNA1I) encoding proteins (CaV3.1–CaV3.3) that differ in their localization as well as in molecular, biophysical, and pharmacological properties. The CACNA1G is a large gene that contains 38 exons and is localized in chromosome 17q22. Only basic characteristics of the CACNA1G gene promoter region have been investigated classifying it as a TATA‐less sequence containing several potential transcription factor‐binding motifs. Here, we cloned and characterized a proximal promoter region and initiated the analysis of transcription factors that control CaV3.1 channel expression using the murine Cacna1g gene as a model. We isolated a ∼1.5 kb 5′‐upstream region of Cacna1g and verified its transcriptional activity in the mouse neuroblastoma N1E‐115 cell line. In silico analysis revealed that this region possesses a TATA‐less minimal promoter that includes two potential transcription start sites and four binding sites for the transcription factor Sp1. The ability of one of these sites to interact with the transcription factor was confirmed by electrophoretic mobility shift assays. Consistent with this, Sp1 over‐expression enhanced promoter activity while siRNA‐mediated Sp1 silencing significantly decreased the level of CaV3.1 protein and reduced the amplitude of whole‐cell T‐type Ca2+ currents expressed in the N1E‐115 cells. These results provide new insights into the molecular mechanisms that control CaV3.1 channel expression. J. Cell. Physiol. 229: 551–560, 2014. © 2013 Wiley Periodicals, Inc.

  January 21, 2014   doi: 10.1002/jcp.24432   open full text
• Induced Neural Lineage Cells as Repair Kits: So Close, Yet So Far away.
  Fahimeh Mirakhori, Bahman Zeynali, Ghasem Hosseini Salekdeh, Hossein Baharvand.
  Journal of Cellular Physiology. November 15, 2013
  Transdifferentiation or direct reprogramming of somatic cells into neural lineage cells has provided an invaluable new tool to advance the regenerative neural medicine. Here, we provide an overview of the various strategies currently available for producing of induced neural lineage cells in vitro as well as the direct reprogramming of neural cells in vivo. We also discussing some of the challenges faced in harnessing the potential of induced neural lineage cells for biomedical applications. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  November 15, 2013   doi: 10.1002/jcp.24509   open full text
• The Dynamic Architectural and Epigenetic Nuclear Landscape: Developing the Genomic Almanac of Biology and Disease.
  Phillip W. L. Tai, Sayyed K. Zaidi, Hai Wu, Rodrigo A. Grandy, Martin M. Montecino, André J. van Wijnen, Jane B. Lian, Gary S. Stein, Janet L. Stein.
  Journal of Cellular Physiology. November 15, 2013
  Compaction of the eukaryotic genome into the confined space of the cell nucleus must occur faithfully throughout each cell cycle to retain gene expression fidelity. For decades, experimental limitations to study the structural organization of the interphase nucleus restricted our understanding of its contributions towards gene regulation and disease. However, within the past few years, our capability to visualize chromosomes in vivo with sophisticated fluorescence microscopy, and to characterize chromosomal regulatory environments via massively‐parallel sequencing methodologies have drastically changed how we currently understand epigenetic gene control within the context of three‐dimensional nuclear structure. The rapid rate at which information on nuclear structure is unfolding brings challenges to compare and contrast recent observations with historic findings. In this review, we discuss experimental breakthroughs that have influenced how we understand and explore the dynamic structure and function of the nucleus, and how we can incorporate historical perspectives with insights acquired from the ever‐evolving advances in molecular biology and pathology. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  November 15, 2013   doi: 10.1002/jcp.24508   open full text
• Ivabradine Protects Against Ventricular Arrhythmias in Acute Myocardial Infarction in the Rat.
  Urszula Mackiewicz, Joseph Y. Gerges, Sandy Chu, Monika Duda, Halina Dobrzynski, Bohdan Lewartowski, Michał Mączewski.
  Journal of Cellular Physiology. November 09, 2013
  Background Ventricular arrhythmias are an important cause of mortality in the acute myocardial infarction (MI). Objective To elucidate effect of ivabradine, pure heart rate (HR) reducing drug, on ventricular arrhythmias within 24 hours after non‐reperfused MI in the rat. Methods ECG was recorded for 24 hours after MI in untreated and ivabradine treated rats and episodes of ventricular tachycardia/fibrillation (VT/VF) were identified. 45 minutes and 24 hours after MI epicardial monophasic action potentials (MAPs) were recorded, cardiomyocyte Ca2+ handling was assessed and expression and function of ion channels were studied. Results Ivabradine reduced average HR by 17%. Combined VT/VF incidence and arrhythmic mortality were higher in MI vs. MI + Ivabradine rats. MI resulted in (1) increase of Ca2+ sensitivity of ryanodine receptors 24 hours after MI; (2) increase of HCN4 expression in the left ventricle (LV) and funny current (IF) in LV cardiomyocytes 24 hours after MI, and (3) dispersion of MAP duration both 45 minutes and 24 hours after MI. Ivabradine partially prevented all these three potential proarrhythmic effects of MI. Conclusions Ivabradine is antiarrhythmic in the acute MI in the rat. Potential mechanisms include prevention of: diastolic Ca2+‐leak from sarcoplasmic reticulum, upregulation of IF current in LV and dispersion of cardiac repolarization. Ivabradine could be an attractive antiarrhythmic agent in the setting of acute MI. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  November 09, 2013   doi: 10.1002/jcp.24507   open full text
• Direct Differentiation of Homogeneous Human Adipose Stem Cells into Functional Hepatocytes by Mimicking Liver Embryogenesis.
  Xueyang Li, Jie Yuan, Weihong Li, Sicheng Liu, Mingxi Hua, Xin Lu, Haiyan Zhang.
  Journal of Cellular Physiology. October 25, 2013
  The potential of adult human adipose tissue stem cells (hASCs) to differentiate into hepatocytes has generated much excitement over the possible use of hASCs in therapeutic applications. An understanding of the molecular mechanisms that underlie the plasticity of hASCs toward hepatocytes will help to make this possibility a reality. Herein, we show that a homogenous population of hASCs characterized by a high level of CD73, CD90, and CD105 express the pluripotent transcription factors OCT4, SOX2, NANOG, and SALL4 under proliferation conditions. A high level of activin A allows for hASCs acquiring the fate of definitive endoderm (DE) cells and expressing the specific transcription factors HEX, FOXA2, SOX17, and GATA4 synchronously. Using a reproducible three‐stage method by mimicking liver embryogenesis, hASCs were directed to differentiate into functional hepatocytes. In the first stage, hASCs were induced to become DE cells by 2 days cultured in serum‐free medium and 3 days of activin A treatment. Next, the presence of fibroblast growth factor (FGF) 4 and bone morphogenetic protein (BMP) 2 in the medium for 5 days induced efficient hepatic differentiation from DE cells. After 10 days of further maturated by the sequential exposure to hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone (DEX), the hASC‐derived hepatocytes expressed mature hepatocytes marker and exhibited functional characterization, including albumin secretion, glycogen storage, urea production, activity of drug transporters, and cytochrome P450 activity. These findings will be useful for the implementation of hASC‐derived hepatocytes in therapeutic purposes, metabolic analyses, drug toxicity screening, and studies of hepatocyte function. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24501   open full text
• 15‐LO/15‐HETE Mediated Vascular Adventitia Fibrosis via p38 MAPK‐Dependent TGF‐β.
  Li Zhang, Yumei Li, Minggang Chen, Xiaojie Su, Dan Yi, Ping Lu, Daling Zhu.
  Journal of Cellular Physiology. October 25, 2013
  15‐Lipoxygenase/15‐hydroxyeicosatetraenoic acid (15‐LO/15‐HETE) is known to modulate pulmonary vascular medial hypertrophy and intimal endothelial cells migration and angiogenesis after hypoxia. However, it is unclear whether 15‐HETE affects the adventitia of the pulmonary arterial wall. We performed immunohistochemistry, adventitia fibrosis, pulmonary artery fibroblasts phenotype and extracellular matrix (ECM) deposition to determine the role of 15‐HETE in hypoxia‐induced pulmonary vascular adventitia remodeling. Our studies showed that O2 deprivation induced adventitia hypertrophy of pulmonary arteries with ECM accumulation in both humans with pulmonary arterial hypertension and hypoxic rats. Hypoxia induced 15‐LO expression in adventitia. With the inhibitor, NDGA depressed the hypoxia induced ECM deposition and 15‐LO production in hypoxic rats. Hypoxia up‐regulated the expression of α‐SMA, type‐Ia collagen and fibronectin in cultured fibroblasts, which seemed to be due to the increased 15‐LO/15‐HETE. Exogenous 15‐HETE mediated the ECM and phenotypic alterations of the fibroblasts as well. The 15‐LO/15‐HETE induced adventitia fibrosis and fibroblasts phenotypic alterations depended on signaling of the transforming growth factor‐β1 (TGF‐β1)/Smad2/3 pathway. P38 mitogen‐activated protein kinase (p38 MAPKs) was likely to mediate 15‐LO induced TGF‐β1 and Smad2/3 activation after hypoxia. The results suggest that adventitia fibrosis is an important event in the hypoxia induced pulmonary arterial remodeling, which relies on 15‐LO/15‐HETE induced p38 MAPK‐dependent TGF‐β1/Smad2/3 intracellular signaling systems. J. Cell. Physiol. 229: 245–257, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24443   open full text
• Neuroprotective Activity of pDING in Response to HIV‐1 Tat.
  Nune Darbinian, Kamel Khalili, Shohreh Amini.
  Journal of Cellular Physiology. October 25, 2013
  Although neurons are not productively infected with HIV‐1, neuronal injury and death are frequently seen in the brains of AIDS patients with neurological and neurocognitive disorders. Evidently, viral proteins including Tat and cellular inflammatory factors released by activated and/or infected microglia, macrophages, and astrocytes contribute to neuronal cell death. Several studies have demonstrated that HIV‐1 associated neuronal cell injury is mediated by dysregulation of signaling pathways that are controlled, in part, by a class of serine/threonine kinases. In this study, we demonstrate that pDING, a novel plant‐derived phosphate binding protein has the capacity to reduce the severity of injury and death caused by HIV‐1 and its neurotoxic Tat protein. We demonstrate that pDING, also called p27SJ/p38SJ, protects cells from the loss of neuronal processes induced by Tat and promotes neuronal outgrowth after Tat‐mediated injury. Further, expression of pDING prevents Tat‐induced oxidative stress and mitochondrial permeability. With its profound phosphatase activity, pDING controls the activity of several kinases including MAPK, Cdk5, and their downstream target protein, MEF2, which is implicated in neuronal cell protection. Our results show that expression of pDING in neuronal cells diminishes the level of hyperphosphorylated forms of Cdk5 and MEF2 caused by Tat and the other neurotoxic agents that are secreted by the HIV‐1 infected cells. These observations suggest that pDING, through its phosphatase activity, has the ability to manipulate the state of phosphorylation and activity of several factors involved in neuronal cell health in response to HIV‐1. J. Cell. Physiol. 229: 153–161, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24392   open full text
• Induction of Mesenchymal Stem Cell Chondrogenesis Through Sequential Administration of Growth Factors Within Specific Temporal Windows.
  Andrew M. Handorf, Wan‐Ju Li.
  Journal of Cellular Physiology. October 25, 2013
  Human mesenchymal stem cells (hMSCs) are capable of differentiating into chondrocyte‐like cells but fail to produce the quality or quantity of cartilage matrix compared to articular chondrocytes using current differentiation protocols. In this study, we aim to improve the chondrogenic differentiation of hMSCs through the sequential administration of multiple growth factors (GFs). We began by looking at differentiating hMSCs' cell surface GF receptor expression every 3 days throughout differentiation using flow cytometry and found that not only was receptor expression dynamic throughout differentiation, but ligand sensitivity was positively correlated with receptor expression, suggesting that differentiating hMSCs may have varying GF requirements depending on their stage of differentiation. We then constructed GF sequences by administering several prochondrogenic GFs singly every 3 days throughout differentiation and assaying the expression of a variety of cartilage‐related genes using qPCR. The resulting chondrocytic phenotype of sequentially induced hMSCs was then compared to that of hMSCs induced under standard culture conditions using qPCR, dimethylmethylene blue assay, and histology. We found that while the initial GF sequence was unable to improve hMSC chondrogenesis, withdrawal of GF treatment at Day 9 of differentiation in pellet culture vastly improved the success of differentiation beyond that induced by TGFβ1 alone. Additional modifications allowed us to further improve chondrogenesis to levels comparable to that obtained by co‐administration of TGFβ1 and BMP7 throughout differentiation. Taken together, we demonstrated the ability to improve the chondrocytic phenotype of differentiated hMSCs through the sequential administration of multiple GFs. J. Cell. Physiol. 229: 162–171, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24428   open full text
• CD43 Signals Prepare Human T Cells to Receive Cytokine Differentiation Signals.
  A.O. Galindo‐Albarrán, O. Ramírez‐Pliego, R.G. Labastida‐Conde, E.I. Melchy‐Pérez, A. Liquitaya‐Montiel, F.R. Esquivel‐Guadarrama, G. Rosas‐Salgado, Y. Rosenstein, M.A. Santana.
  Journal of Cellular Physiology. October 25, 2013
  T cells are increasingly used for passive immunotherapy and bone marrow transplantation. Proper ex‐vivo management of the cells is important for the desired therapeutic effects. For differentiation into effector cells of the Th1 and Th2 phenotypes, T‐cells require signals from IFNγ and IL‐4, respectively. Naïve cells have an extremely low expression of the specific receptors that recognize these cytokines, indicating that in order to differentiate, cells need to perceive other signals that will enable them to sense the cytokine milieu. CD43 has been proposed as one of the molecules that make the initial contacts with antigen presenting cells. We report here that in cord blood, adult naïve and total human T cells, CD43 signals induced the expression of both IFNγ and IL‐4 receptors, mediate their capping, increased their signaling and augmented differentiation mediated by these receptors. CD43 signals also stimulated the expression of IFNγ and in neonatal cells that of IL‐4 as well. These data demonstrate an important role for CD43 signals in T‐cell preparedness for differentiation into effector cells. J. Cell. Physiol. 229: 172–180, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24430   open full text
• TMEM16E (GDD1) Exhibits Protein Instability and Distinct Characteristics in Chloride Channel/Pore Forming Ability.
  Ta To Tran, Kei Tobiume, Chikara Hirono, Shinichi Fujimoto, Kuniko Mizuta, Kazumi Kubozono, Hiroshi Inoue, Mitsuo Itakura, Makoto Sugita, Nobuyuki Kamata.
  Journal of Cellular Physiology. October 25, 2013
  TMEM16E/GDD1 has been shown to be responsible for the bone‐related late‐onset disease gnathodiaphyseal dysplasia (GDD), with the dominant allele (TMEM16Egdd) encoding a missense mutation at Cys356. Additionally, several recessive loss‐of‐function alleles of TMEM16E also cause late‐onset limb girdle muscular dystrophy. In this study, we found that TMEM16E was rapidly degraded via the proteasome pathway, which was rescued by inhibition of the PI3K pathway and by the chemical chaperone, sodium butyrate. Moreover, TMEM16Egdd exhibited lower stability than TMEM16E, but showed similar propensity to be rescued. TMEM16E did not exhibit cell surface calcium‐dependent chloride channel (CaCC) activity, which was originally identified in TMEM16A and TMEM16B, due to their intracellular vesicle distribution. A putative pore‐forming domain of TMEM16E, which shared 39.8% similarity in 98 amino acids with TMEM16A, disrupted CaCC activity of TMEM16A via domain swapping. However, the Thr611Cys mutation in the swapped domain, which mimicked conserved cysteine residues between TMEM16A and TMEM16B, reconstituted CaCC activity. In addition, the GDD‐causing cysteine mutation made in TMEM16A drastically altered CaCC activity. Based on these findings, TMEM16E possesses distinct function other than CaCC and another protein‐stabilizing machinery toward the TMEM16E and TMEM16Egdd proteins should be considered for the on‐set regulation of their phenotypes in tissues. J. Cell. Physiol. 229: 181–190, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24431   open full text
• RECK Inhibits Stemness Gene Expression and Tumorigenicity of Gastric Cancer Cells by Suppressing ADAM‐Mediated Notch1 Activation.
  Kun‐Jing Hong, Deng‐Chyang Wu, Kuang‐Hung Cheng, Li‐Tzong Chen, Wen‐Chun Hung.
  Journal of Cellular Physiology. October 25, 2013
  The Reversion‐inducing Cysteine‐rich Protein with Kazal Motifs (RECK) gene encodes a membrane‐anchored glycoprotein that exhibits strong inhibitory activity against various matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinase 10 (ADAM10). RECK functions as a tumor suppressor by inhibiting migration, invasion, and angiogenesis. However, whether RECK can modulate the stem‐like phenotypes of cancer cells is not known. In this study, we demonstrate that RECK is down‐regulated in gastric cancer cells and is further reduced in CD133‐positive cancer stem‐like cells. Ectopic expression of RECK induces down‐regulation of the expression of stemness genes including Sox2, Oct4, and Nanog and the cancer stem cell marker CD133. Treatment of DAPT (a γ‐secretase inhibitor) or TAPI‐2 (a hydroxamate‐based inhibitor of MMPs, tumor necrosis factor α converting enzyme and ADAM17) reduces Notch1 shedding and activation which results in attenuation of stemness genes and CD133. Our data show that ADAM10 and ADAM17 are co‐pulled down by RECK suggesting a physical interaction between RECK and ADAMs on cell surface. In addition, RECK suppresses sphere formation and sphere size of CD133‐positive gastric cancer cells. Overexpression of Notch intracellular domain (NICD) or ADAM17 effectively reverse the inhibitory effect of RECK in CD133‐positive cells. More importantly, RECK reduces tumorigenic activity of CD133‐positive cells in vivo. Conversely, knockdown of RECK in non‐tumorigenic GI2 cells increases stemness and CD133 expression and sphere forming ability. Collectively, these results indicate that RECK represses stemness gene expression and stem‐like properties by inhibiting ADAM‐mediated Notch1 shedding and activation. J. Cell. Physiol. 229: 191–201, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24434   open full text
• Inhibition of Wnt/β‐Catenin Signaling Promotes Engraftment of Mesenchymal Stem Cells to Repair Lung Injury.
  Zhaorui Sun, Xuemin Gong, Huiming Zhu, Cong Wang, Xiaomeng Xu, Di Cui, Weiping Qian, Xiaodong Han.
  Journal of Cellular Physiology. October 25, 2013
  We sought to explore the treatment effects and the repair mechanisms of bone marrow derived mesenchymal stem cells (MSCs) during HCl‐induced acute lung injury (ALI). MSCs were delivered through the tail veins of rats 24 h after intranasal instillation of HCl. The results showed that MSCs did not ameliorate the histopathologic changes of ALI and pulmonary fibrosis. We found that the activated Wnt/β‐catenin signaling may regulate the differentiation of MSCs and is associated with lung fibroblasts activation, pulmonary fibrosis and tissue repair process in ALI rats. Immunofluorescence and histology analysis indicated that activated canonical Wnt/β‐catenin signaling induced most MSCs to differentiate into myofibroblasts or fibroblasts in vivo. However, inhibition of Wnt/β‐catenin signaling by Dickkopf‐1 (DKK1) promotes epithelial differentiation of MSCs induced by native alveolar epithelial cells which are beneficial to repair the injured lung epithelium. Inhibition of Wnt/β‐catenin signaling after MSCs transplantation ameliorated pulmonary fibrosis and improved pulmonary function which attenuated the lung injury. In vitro study, activation of the Wnt/β‐catenin signaling stimulated MSCs to express myofibroblasts markers, which was attenuated by DKK1. Furthermore, Wnt3α activated Wnt/β‐catenin signaling in lung fibroblasts to enhance the expression of collagen I, vimentin and α‐smooth muscle actin, but DKK1 attenuated these proteins expression. These findings demonstrated that canonical Wnt/β‐catenin signaling plays a key role in regulating differentiation of MSCs in vivo or in vitro and the pathogenesis of fibrotic diseases. Our study suggested that inhibition of abnormal activated Wnt/β‐catenin signaling would promote MSCs epithelial differentiation to repair lung injury and reduce pulmonary fibrosis. J. Cell. Physiol. 229: 213–224, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24436   open full text
• BKCa and hEag1 Channels Regulate Cell Proliferation and Differentiation in Human Bone Marrow‐Derived Mesenchymal Stem Cells.
  Ying‐Ying Zhang, Jianbo Yue, Hui Che, Hai‐Ying Sun, Hung‐Fat Tse, Gui‐Rong Li.
  Journal of Cellular Physiology. October 25, 2013
  Human bone marrow‐derived mesenchymal stem cells (MSCs) serve as a reservoir for the continuous renewal of various mesenchymal tissues; however, cellular physiology of ion channels is not fully understood. The present study investigated potential roles of large‐conductance Ca2+‐activated potassium (BKCa) channels and ether‐à‐go‐go potassium (hEag1 or Kv10.1) channels in regulating cell proliferation and differentiation in human MSCs. We found that inhibition of BKCa with paxilline or hEag1 with astemizole, or knockdown of BKCa with shRNAs targeting KCa1.1 or hEag1 channels with shRNAs targeting KCNH1 arrested the cells at G0/G1 phase. In addition, silencing BKCa or hEag1 channels significantly reduced adipogenic differentiation with decrease of lipid accumulation and expression of the adipocyte marker PPARγ, and decreased osteogenic differentiation with reduction of mineral precipitation and osteocalcin. These effects were accompanied with a reduced cyclin D1, cyclin E, p‐ERK1/2, and p‐Akt. Our results demonstrate that BKCa and hEag1 channels not only regulate cell proliferation, but also participate in the adipogenic and osteogenic differentiations in human MSCs, which indicates that BKCa and hEag1 channels may be essential in maintaining bone marrow physiological function and bone regeneration. J. Cell. Physiol. 229: 202–212, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24435   open full text
• A Commentary on iPS Cells: Potential Applications in Autologous Transplantation, Study of Illnesses and Drug Screening.
  Gaetano Romano, Fátima Morales, Ignazio R. Marino, Antonio Giordano.
  Journal of Cellular Physiology. October 25, 2013
  Undoubtedly, the focus of the field of stem cell research is predominantly aimed at the artificial reprogramming of human somatic cells for the production of induced pluripotent stem (iPS) cells. This relatively new technology may circumvent the ethical issues of using human embryonic stem (hES) cells for the potential applications in cell replacement therapy. Besides such ethical issues, iPS cell technology offers the advantage of obtaining patient‐derived tissues and/or cells, which may be utilized for autologous transplantation and tissue regeneration, investigation of a variety of human illnesses and for the screening of new drugs. The field of stem cell research has placed a major emphasis in understanding the genetic and epigenetic codes for pluripotency, in order to control and optimize autologous transplantation techniques and avoid teratoma formation. J. Cell. Physiol. 229: 148–152, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24437   open full text
• PAP/HIP Protein Is an Obesogenic Factor.
  Veronique Secq, Cecilia Mallmann, Meritxell Gironella, Belen Lopez, Daniel Closa, Stéphane Garcia, Laurence Christa, Giuseppe Montalto, Nelson Dusetti, Juan L. Iovanna.
  Journal of Cellular Physiology. October 25, 2013
  In this article we report the obesogenic role of the acute phase protein PAP/HIP. We found that the transgenic TgPAP/HIP mice develop spontaneous obesity under standard nutritional conditions, with high levels of glucose, leptin, and LDL and low levels of triglycerides and HDL in blood. Accordingly, PAP/HIP‐deficient mice are skinny under standard nutritional conditions. We also found that expression of PAP/HIP is induced in intestinal epithelial cells in response to gavage with olive oil and this induction is AG490 sensitive. We demonstrated that incubation of 3T3‐L1 preadipocytes with a low concentration as 1 ng/ml of recombinant PAP/HIP results in accelerated BrdU incorporation in vitro. PAP/HIP‐dependent adipocytes growth is sensitive to the MEK inhibitor U0126. Finally, patients with severe obesity present higher blood levels of PAP/HIP than non‐obese control individuals. Altogether our data suggest that PAP/HIP could be a mediator of fat tissue development, released by the intestine and induced by the presence of food into the gut. J. Cell. Physiol. 229: 225–231, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24438   open full text
• Induction of Dopaminergic Neurons From Human Wharton's Jelly Mesenchymal Stem Cell by Forskolin.
  Emanuela Paldino, Carlo Cenciarelli, Adele Giampaolo, Luisa Milazzo, Mario Pescatori, Hamisa Jane Hassan, Patrizia Casalbore.
  Journal of Cellular Physiology. October 25, 2013
  The purpose of this study was to investigate the Wharton's jelly mesenchymal stem cells differentiation ability toward neuronal fate. Human Wharton's jelly mesenchymal stem cells (hWJMSC) have been isolated from human umbilical cord of full‐term births and characterized by flow cytometry analysis for their stem mesenchymal properties through specific surface markers expression (CD73, CD90, and CD105). hWJMSC mesodermal lineage differentiation ability and karyotype analysis were assessed. The trans‐differentiation of hWJMSC into neural lineage was investigated in presence of forskolin, an agent known to increase the intracellular levels of cAMP. A molecular profile of differentiated hWJMSC was performed by microarray technology which revealed 1,532 statistically significant modulated genes respect to control cells. Most of these genes are mainly involved in functional neuronal signaling pathways and part of them are specifically required for the neuronal dopaminergic induction. The acquisition of the dopaminergic phenotype was evaluated via immunocytochemistry and Western blot analysis revealed the significant induction of Nurr1, NeuroD1, and TH proteins expression in forskolin‐induced hWJMSC. Moreover, the treatment with forskolin promoted, in hWJMSC, a strong upregulation of the neurotrophin Trk receptors related to the high release of brain‐derived neurotrophic factor. Taken together these findings show that hWJMSC may be represent an optimal therapeutic strategy for neurological diseases. J. Cell. Physiol. 229: 232–244, 2014. © 2013 Wiley Periodicals, Inc.

  October 25, 2013   doi: 10.1002/jcp.24442   open full text
• M2 Receptors Exert Analgesic Action on DRG Sensory Neurons by Negatively Modulating VR1 Activity.
  Federica De Angelis, Sara Marinelli, Bernard Fioretti, Luigi Catacuzzeno, Fabio Franciolini, Flaminia Pavone, Ada Maria Tata.
  Journal of Cellular Physiology. October 24, 2013
  The peripheral application of the M2 cholinergic agonist arecaidine on sensory nerve endings shows antinociceptive properties. In this work we analyze in vitro, the mechanisms downstream M2 receptor activation causing the analgesic effects, and in vivo the effects produced by M2 agonist arecaidine administration on nociceptive responses in a murine model of NGF‐induced pain. Cultured DRG neurons treated with arecaidine showed a decreased level of VR1 and SP transcripts. Conversely we found an increased expression of VR1 and SP transcripts in DRG from M2/M4‐/‐ mice compared to WT and M1‐/‐ mice, confirming the inhibitory effect in particular of M2 receptors on SP and VR1 expression. Patch‐clamp experiments in the whole‐cell configuration showed that arecaidine treatment caused a reduction of the fraction of capsaicin‐responsive cells, without altering the mean capsaicin‐activated current in responsive cells. We also demonstrated that arecaidine prevents PKCϵ translocation to the plasma membrane after inflammatory agent stimulation, mainly in medium‐small sensory neurons. Finally, in mice we have observed that intraperitoneal injection of arecaidine reduces VR1 expression blocking hyperalgesia and allodynia caused by NGF intraplantar administration. In conclusion, our data demonstrate that in vivo M2 receptor activation induces desensitization to mechanical and heat stimuli by a down‐regulation of VR1 expression and by the inhibition of PKCϵ activity hindering its translocation to the plasma membrane, as suggested by in vitro experiments. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 24, 2013   doi: 10.1002/jcp.24499   open full text
• Stable Genetic Alterations of β‐Catenin and ROR2 Mediated by Lentiviral Vectors Regulate the Wnt Pathway and Affect the Fate of MSCs.
  Shi‐xia Cai, Ai‐ran Liu, Hong‐li He, Qi‐hong Chen, Yi Yang, Feng‐mei Guo, Ying‐zi Huang, Ling Liu, Hai‐bo Qiu.
  Journal of Cellular Physiology. October 24, 2013
  The Wnt pathways have been shown to be critical for the fate of Mesenchymal stem cells (MSCs) in vitro, but their roles in MSCs in vivo remain poorly characterized due to the lack of stable alterations in their signaling. In the present study, we constructed long‐term and stable mMSCs lines with activated and inactivated β‐catenin (the key molecule of the canonical Wnt signaling pathway) or ROR2 (the key molecule of the noncanonical Wnt5a/ROR2 signaling pathway) modifications with lentiviral vectors. We found that the transduction efficiencies mediated by the lentiviral vectors were 92.61%‐97.04% and were maintained over 20 passages of mMSCs. Transfection by lentiviral vectors not only regulated the mRNA and protein expression of β‐catenin or ROR2 but also regulated nuclear β‐catenin accumulation or the Wnt5a/JNK and Wnt5a/PKC pathways belonging to the canonical Wnt and noncanonical Wnt5a/ROR2 pathways, respectively. β‐catenin or ROR2 gene overexpression promoted mMSC proliferation, migration and differentiation into osteoblasts, while inhibiting the adipogenic differentiation of mMSCs. In contrast, inactivation of the β‐catenin or ROR2 genes resulted in the opposite effects. Therefore, these results confirm that lentiviral vector transduction can facilitate sustained and efficient gene modification of the Wnt pathway in mMSCs. This study provides a method to investigate the effects of the Wnt pathway on the fate of mMSCs in vivo and for the further improvement of MSC‐based therapies. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 24, 2013   doi: 10.1002/jcp.24500   open full text
• GKN2 Contributes to the Homeostasis of Gastric Mucosa by Inhibiting GKN1 Activity.
  Olga Kim, Jung Hwan Yoon, Won Suk Choi, Hassan Ashktorab, Duane T. Smoot, Suk Woo Nam, Jung Young Lee, Won Sang Park.
  Journal of Cellular Physiology. October 22, 2013
  Gastrokine 1 (GKN1) plays an important role in maintaining gastric mucosa integrity. Here, we investigated whether gastrokine 2 (GKN2) contributes to the homeostasis of gastric epithelial cells by regulating GKN1 activity. We analyzed cell viability, proliferation, and death in AGS cells transfected with GKN1, GKN2, GKN1 plus GKN2 using MTT, BrdU incorporation, and apoptosis assays, respectively. In addition, the expression levels of the cell cycle‐ and apoptosis‐related proteins, miR‐185, DNMT1, and EZH2 were determined. We also compared the expression of GKN1, GKN2, and CagA in 50 non‐neoplastic gastric mucosae and measured GKN2 expression in 169 gastric cancers by immunohistochemistry. GKN2 inhibited anti‐proliferative and pro‐apoptotic activities, miR‐185 induction, and anti‐epigenetic modifications of GKN1. There was a positive correlation between GKN1 and GKN2 expression (P = 0.0074), and the expression of GKN1, but not GKN2, was significantly lower in H. pylori CagA‐positive gastric mucosa (P = 0.0013). Interestingly, ectopic GKN1 expression in AGS cells increased GKN2 mRNA and protein expression in a time‐dependent manner (P = 0.01). Loss of GKN2 expression was detected in 126 (74.6%) of 169 gastric cancers by immunohistochemical staining and was closely associated with GKN1 expression and differentiation of gastric cancer cells (P = 0.0002 and P = 0.0114, respectively). Overall, our data demonstrate that in the presence of GKN2, GKN1 loses its ability to decrease cell proliferation, induce apoptosis, and inhibit epigenetic alterations in gastric cancer cells. Thus, we conclude that GKN2 may contribute to the homeostasis of gastric epithelial cells by inhibiting GKN1 activity. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 22, 2013   doi: 10.1002/jcp.24496   open full text
• MicroRNA‐200b Stimulates Tumour Growth in TGFBR2‐Null Colorectal Cancers by Negatively Regulating p27/kip1.
  Yuxuan Fu, Xianghua Liu, Ningtian Zhou, Lijian Du, Yu Sun, Xiang Zhang, Yingbin Ge.
  Journal of Cellular Physiology. October 22, 2013
  Colorectal cancer (CRC) remains the most common malignancy worldwide. TGF‐β1 is often overexpressed in late stages of colorectal carcinogenesis and promotes tumour growth and metastasis. Several reports have verified that the loss of functional TGFBRII expression contributed to escape the tumour suppressor activity of TGF‐β1 and that the epithelial‐to‐mesenchymal transition (EMT) responded to TGF‐β1 involved in tumour invasion and metastasis. However, the mechanisms by which TGF‐β1 confers a growth advantage to TGFBRII‐null colorectal cancer cells have not been elucidated. MicroRNAs (miRNAs) are post‐transcriptional inhibitory regulators of gene expression that act by directly binding complementary mRNA and are key determinants of cancer initiation and progression. In this study, we revealed a role for miR‐200b in colorectal cancer. MiR‐200b was highly expressed in TGFBRII‐null tumour tissues and colorectal cancer cell lines and positively correlated with cell proliferation in tumour tissues and cell lines. In contrast, decreasing the miR‐200b level in TGFBRII‐null cells suppressed cell proliferation and cell cycle progression. Furthermore, in vivo studies also suggested a stimulating effect of miR‐200b on TGFBRII‐null cell‐derived xenografts. CDKN1B (p27/kip1) and RND3 (RhoE) have miR‐200b binding sequences within their 3' untranslated regions and were confirmed to be direct targets of miR‐200b using fluorescent reporter assays. Meanwhile, CDKN1B (p27/kip1) played a role in miR‐200b‐stimulated TGFBR‐null CRC. This study suggests that miR‐200b plays a tumour‐promoting role by targeting CDKN1B (p27/kip1) in CRCs. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 22, 2013   doi: 10.1002/jcp.24497   open full text
• Non‐genomic estrogen/estrogen receptor α promotes cellular malignancy of immature ovarian teratoma in vitro.
  Yao‐Chin Hung, Wei‐Chun Chang, Lu‐Min Chen, Ying‐Yi Chang, Ling‐Yu Wu, Wei‐Min Chung, Tze‐Yi Lin, Liang‐Chi Chen, Wen‐Lung Ma.
  Journal of Cellular Physiology. October 19, 2013
  Malignant immature ovarian teratomas (IOT) most often occur in women of reproductive age. It is unclear, however, what roles estrogenic signaling plays in the development of IOT. In this study, we examined whether estrogen receptors (ERα and β) promote the cellular malignancy of IOT. Estradiol (E2), PPT (propylpyrazole), and DPN (diarylpropionitrile) (ERα‐ and β‐specific agonists, respectively), as well as ERα‐ or ERβ‐specific short hairpin (sh)RNA were applied to PA‐1 cells, a well‐characterized IOT cell line. Cellular tumorigenic characteristics, e.g., cell migration/invasion, expression of the cancer stem/progenitor cell marker CD133, and evidence for epithelial‐mesenchymal transition (EMT) were examined. In PA‐1 cells that expressed ERα and ERβ, we found that ERα promoted cell migration and invasion. We also found that E2/ERα signaling altered cell behavior through non‐classical transactivation function. Our data show non‐genomic E2/ERα activations of focal adhesion kinase‐Ras homolog gene family member A (FAK‐RhoA) and ERK governed cell mobility capacity. Moreover, E2/ERα signaling induces EMT and overexpression of CD133 through upregulation micro‐RNA 21 (miR21; IOT stem/progenitor promoter), and ERK phosphorylations. Furthermore, E2/ERα signaling triggers a positive feedback regulatory loop within miR21 and ERK. At last, expression levels of ERα, CD133, and EMT markers in IOT tissue samples were examined by immunohistochemistry. We found that cytosolic ERα was co‐expressed with CD133 and mesenchymal cell markers but not epithelial cell markers. In conclusion, estrogenic signals exert malignant transformation capacity of cancer cells, exclusively through non‐genomic regulation in female germ cell tumors. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 19, 2013   doi: 10.1002/jcp.24495   open full text
• Platelet‐derived growth factor BB mimics serum‐induced dispersal of pancreatic epithelial cell clusters.
  Sahar Hiram‐Bab, Liora S. Katz, Hagit Shapira, Judith Sandbank, Marvin C. Gershengorn, Yoram Oron.
  Journal of Cellular Physiology. October 15, 2013
  We showed previously that proliferating human islet‐derived de‐differentiated cells (DIDs) exhibit many characteristics of mesenchymal stem cells. Dispersed DIDs can be induced by serum deprivation to undergo mesenchymal‐to‐epithelial transition and aggregate into epithelial cell clusters (ECCs). Conversely, ECCs can be induced to disperse and undergo epithelial‐to‐mesenchymal transition (EMT) by re‐addition of mammalian sera. In this study, we show that platelet‐derived growth factor BB (PDGF‐BB) mimics and mediates serum‐induced ECCs' dispersal accompanied by accumulation of cytoplasmic β‐catenin and a decrease in the levels of insulin and glucagon mRNAs. Moreover, we show that PDGF‐BB‐induced dispersal of ECCs is a more general phenomenon that occurs also with bone marrow mesenchymal stem cells (BM‐MSCs) and dermal fibroblasts (DFs). In DIDs, BM‐MSCs and DFs, PDGF decreased the levels of DKK1 mRNA, suggesting involvement of the Wnt signaling pathway. PDGF‐BB stimulated a significant increase in S473 phosphorylation of Akt and the PI3K specific inhibitor (PIP828) partially inhibited PDGF‐BB‐induced ECC dispersal. Lastly, the PDGF‐receptor (PDGF‐R) antagonist JNJ‐10198409 inhibited both PDGF‐BB – and serum‐induced ECC dispersal. Epidermal growth factor (EGF), which shares most of the PDGF signaling pathway, did not induce dispersal and only weakly stimulated Akt phosphorylation. Our data suggest that PDGF‐BB mediates serum‐induced DIDs dispersal, correlated with the activation of the PI3K‐Akt pathway. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 15, 2013   doi: 10.1002/jcp.24493   open full text
• Angiogenin stimulates ribosomal RNA transcription by epigenetic activation of the ribosomal DNA promoter.
  Jinghao Sheng, Wenhao Yu, Xiangwei Gao, Zhengping Xu, Guo‐Fu Hu.
  Journal of Cellular Physiology. October 03, 2013
  Angiogenin (ANG) undergoes nuclear translocation and promotes ribosomal RNA (rRNA) transcription thereby enhancing cell growth and proliferation. However, the mode of action of ANG in stimulating rRNA transcription is unclear. Here, we show that ANG enhances the formation of RNA polymerase I (Pol I) pre‐initiation complex at the ribosomal DNA (rDNA) promoter. ANG binds at the upstream control element (UCE) of the promoter and enhances promoter occupancy of RNA Pol I as well as the selectivity factor SL1 components TAFI48 and TAFI110. We also show that ANG increases the number of actively transcribing rDNA by epigenetic activation through promoter methylation and histone modification. ANG binds to histone H3, inhibits H3K9 methylation, and activates H3K4 methylation as well as H4 acetylation at the rDNA promoter. These data suggest that one of the mechanisms by which ANG stimulates rRNA transcription is through an epigenetic activation of rDNA promoter. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  October 03, 2013   doi: 10.1002/jcp.24477   open full text
• Retromer promotes immune quiescence by suppressing Spätzle‐Toll pathway in Drosophila.
  Bo Zhou, Eun‐Young Yun, Lorraine Ray, Jia You, Y. Tony Ip, Xinhua Lin.
  Journal of Cellular Physiology. September 24, 2013
  The Toll and Toll‐Like Receptor signaling pathways are evolutionarily conserved pathways that regulate innate immunity in insects and mammals. While efforts have been made to clarify the signal transduction events that occur during infection, much less is known about the components that maintain immune quiescence. Here we show that retromer, an intracellular protein complex known for regulating vesicle trafficking, functions in modulating the Toll pathway in Drosophila melanogaster. In mutant animals lacking retromer function, the Toll pathway but not JAK‐STAT or IMD pathway is activated, triggering both cellular and humoral responses. Genetic epistasis and clonal analysis suggest that retromer regulates a component that acts upstream of Toll. Our data further show that in the mutant the Toll ligand Spätzle has a processing pattern similar to that of after infection. Together, the results suggest a novel function of retromer in regulating Toll pathway and innate immunity at a step that modulates ligand processing or activity. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 24, 2013   doi: 10.1002/jcp.24472   open full text
• G protein‐coupled receptors and adipogenesis: A focus on adenosine receptors.
  Anna Eisenstein, Katya Ravid.
  Journal of Cellular Physiology. September 24, 2013
  G‐protein coupled receptors (GPCRs) are a large family of proteins that coordinate extracellular signals to produce physiologic outcomes. Adenosine receptors (AR) are one class of GPCRs that have been shown to regulate functions as diverse as inflammation, blood flow, and cellular differentiation. Adenosine signals through four GPCRs that either inhibit (A1AR and A3AR) or activate (A2aAR and A2bAR) adenylyl cyclase. This review will focus on the role of GPCRs, and in particular, adenosine receptors, in adipogenesis. Preadipocytes differentiate to mature adipocytes as the adipose tissue expands to compensate for the consumption of excess nutrients. These newly generated adipocytes contribute to maintaining metabolic homeostasis. Understanding the key drivers of this differentiation process can aid the development of therapeutics to combat the growing obesity epidemic and associated metabolic consequences. Although much literature has covered the transcriptional events that culminate in the formation of an adipocyte, less focus has been on receptor‐mediated extracellular signals that direct this process. This review will highlight GPCRs and their downstream messengers as significant players controlling adipocyte differentiation. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 24, 2013   doi: 10.1002/jcp.24473   open full text
• The BRG1 Chromatin Remodeler Regulates Widespread Changes in Gene Expression and Cell Proliferation During B Cell Activation.
  Darcy W. Holley, Beezly S. Groh, Glenn Wozniak, Dallas R. Donohoe, Wei Sun, Virginia Godfrey, Scott J. Bultman.
  Journal of Cellular Physiology. September 23, 2013
  Widespread changes in gene expression underlie B cell development and activation, yet our knowledge of which chromatin‐remodeling factors are essential is limited. Here, we demonstrate that the BRG1 catalytic subunit of SWI/SNF complexes was dispensable for murine B cell development but played an important, albeit selective, role during activation. Although BRG1 was dispensable for CD69 induction and differentiation into plasma cells based on the ability of mutant B cells to undergo hypertrophy and secrete IgM antibodies, it was required for robust cell proliferation in response to activation. Accordingly, BRG1 was required for only ∼100 genes to be expressed at normal levels in naïve B cells but >1,000 genes during their activation. BRG1 upregulated fivefold more genes than it downregulated, and the toll‐like receptor pathway and JAK/STAT cytokine‐signaling pathways were particularly dependent on BRG1. The importance of BRG1 in B cell activation was underscored by the occurrence of opportunistic Pasteurella infections in conditionally mutant mice. B cell activation has long served as a model of inducible gene expression, and the results presented here identify BRG1 as a chromatin‐remodeling factor that upregulates the transcriptome of B cells during their activation to promote rapid cell proliferation and to mount an effective immune response. J. Cell. Physiol. 229: 44–52, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24414   open full text
• TRPM7 Channels Regulate Proliferation and Adipogenesis in 3T3‐L1 Preadipocytes.
  Kui‐Hao Chen, Xiao‐Hui Xu, Yi Liu, Yan Hu, Man‐Wen Jin, Gui‐Rong Li.
  Journal of Cellular Physiology. September 23, 2013
  Transient receptor potential melastatin‐7 (TRPM7) channels are involved in many cellular physiological and pathological processes. The present study was designed to investigate the expression of TRPM7 channels and the potential role in regulating cell proliferation and adipogenesis in 3T3‐L1 preadipocytes with approaches of whole‐cell patch voltage‐clamp, molecular biology, cell proliferation, adipogenesis, etc. We found that a TRPM7‐like current was recorded with Mg2+‐free pipette solution in 3T3‐L1 preadipocytes, and the current was inhibited by intercellular free Mg2+. The TRPM7‐like current was potentiated by acidic pH and inhibited by 2‐aminoethoxydiphenyl borate (2‐APB). RT‐PCR, Western blot and immunocytochemistry revealed that gene and protein of TRPM7 channels were abundant in 3T3‐L1 preadipocytes. Blockade of TRPM7 channels with 2‐APB inhibited cell proliferation in 3T3‐L1 cells. In addition, knockdown of TRPM7 with specific siRNA inhibited both proliferation and adipogenesis. The present study demonstrates for the first time that TRPM7 channels regulate cell cycle and adipogenesis of 3T3‐L1 preadipocytes. J. Cell. Physiol. 229: 60–67, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24417   open full text
• Chronic Nicotine Exposure Systemically Alters MicroRNA Expression Profiles During Post‐Embryonic Stages in Caenorhabditis elegans.
  Faten A Taki, Xiaoping Pan, Baohong Zhang.
  Journal of Cellular Physiology. September 23, 2013
  Tobacco smoking is associated with many diseases. Addiction is of the most notorious tobacco‐related syndrome and is mainly attributed to nicotine. In this study, we employed Caenorhabditis elegans as a biological model to systemically investigate the effect of chronic nicotine exposure on microRNA (miRNA) expression profile and their regulated biochemical pathways. Nicotine treatment (20 µM and 20 mM) was limited to the post‐embryonic stage from L1 to L4 (∼31 h) period after which worms were collected for genome‐wide miRNA profiling. Our results show that nicotine significantly altered the expression patterns of 40 miRNAs. The effect was proportional to the nicotine dose and was expected to have an additive, more robust response. Based on pathway enrichment analyses coupled with nicotine‐induced miRNA patterns, we inferred that miRNAs as a system mediates “regulatory hormesis”, manifested in biphasic behavioral and physiological phenotypes. We proposed a model where nicotine addiction is mediated by miRNAs' regulation of fos‐1 and is maintained by epigenetic factors. Thus, our study offers new insights for a better understanding of the sensitivity of early developmental stages to nicotine. J. Cell. Physiol. 229: 78–89, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24419   open full text
• A Role for the Chemokine Receptor CCR6 in Mammalian Sperm Motility and Chemotaxis.
  Pedro Caballero‐Campo, Mariano G. Buffone, Fabian Benencia, José R. Conejo‐García, Paolo F. Rinaudo, George L. Gerton.
  Journal of Cellular Physiology. September 23, 2013
  Although recent evidence indicates that several chemokines and defensins, well‐known as inflammatory mediators, are expressed in the male and female reproductive tracts, the location and functional significance of chemokine networks in sperm physiology and sperm reproductive tract interactions are poorly understood. To address this deficiency in our knowledge, we examined the expression and function in sperm of CCR6, a receptor common to several chemoattractant peptides, and screened several reproductive tract fluids for the presence of specific ligands. CCR6 protein is present in mouse and human sperm and mainly localized in the sperm tail with other minor patterns in sperm from mice (neck and acrosomal region) and men (neck and midpiece regions). As expected from the protein immunoblotting and immunofluorescence results, mouse Ccr6 mRNA is expressed in the testis. Furthermore, the Defb29 mRNA encoding the CCR6 ligand, β‐defensin DEFB29, is expressed at high levels in the epididymis. As determined by protein chip analysis, several chemokines (including some that act through CCR6, such as CCL20/MIP‐3α (formerly macrophage inflammatory protein 3α) and protein hormones were present in human follicular fluid, endometrial secretions, and seminal plasma. In functional chemotaxis assays, capacitated human sperm exhibited a directional movement towards CCL20, and displayed modifications in motility parameters. Our data indicate that chemokine ligand/receptor interactions in the male and female genital tracts promote sperm motility and chemotaxis under non‐inflammatory conditions. Therefore, some of the physiological reactions mediated by CCR6 ligands in male reproduction extend beyond a pro‐inflammatory response and might find application in clinical reproduction and/or contraception. J. Cell. Physiol. 229: 68–78, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24418   open full text
• High HMGA2 Expression and High Body Mass Index Negatively Affect the Prognosis of Patients With Ovarian Cancer.
  Daniela Califano, Sandro Pignata, Nunzia Simona Losito, Alessandro Ottaiano, Stefano Greggi, Veronica De Simone, Sabrina Cecere, Concetta Aiello, Francesco Esposito, Alfredo Fusco, Gennaro Chiappetta.
  Journal of Cellular Physiology. September 23, 2013
  HMGA2 is a small, non‐histone, chromatin‐associated protein with a key role in tumorigenesis and adipogenesis. Indeed, HMGA2 overexpression has been frequently detected in several malignant neoplasms and inhibition of its expression prevents thyroid cell transformation. Moreover, HMGA2 null mice show a pigmy phenotype with a great reduction in fat tissue. To investigate whether HMGA2 expression correlates with clinico‐pathological parameters and patient outcome, immunohistochemical analysis of HMGA2 expression was performed in ovarian cancer specimens from 117 patients. HMGA2 overexpression was found in 39% of the cases and, interestingly, positively correlated with the body mass index (BMI). Moreover, high BMI (≥25 kg/m2) and high HMGA2 expression/BMI combined evaluation predicted shorter disease‐free survival. High BMI (≥25 kg/m2), high expression of HMGA2 and high HMGA2 expression/BMI combined evaluation predicted shorter overall survival. In multivariate analysis, the concomitant high expression of HMGA2 and high BMI (≥25 kg/m2) was an independent prognostic factor. Finally, the BMI (≥25 kg/m2) negatively correlated with the patient response to chemotherapy (P = 0.039). Therefore, the data reported herein suggest that the combined evaluation of HMGA2 expression and obesity assessed through BMI can be considered a marker of poor prognosis in patients affected by ovarian carcinoma. J. Cell. Physiol. 229: 53–59, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24416   open full text
• Vascular Endothelial Growth Factor Does not Accelerate Endothelial Differentiation of Human Mesenchymal Stem Cells.
  Richard J. Galas, Julie C. Liu.
  Journal of Cellular Physiology. September 23, 2013
  For clinical applications of engineered vascular replacements, endothelial cells may not be available in sufficient quantities due to limited harvesting sites and slow in vitro expansion rates. Soluble vascular endothelial growth factor (VEGF) is often added to differentiate mesenchymal stem cells (MSCs) into endothelial cells; however, recent studies demonstrate that VEGF is not required to upregulate endothelial markers. In contrast to previous assumptions, this study demonstrates that exogenous VEGF does not enhance or accelerate the upregulation of common endothelial markers during endothelial differentiation of human MSCs. MSCs were cultured at confluence for up to 3 weeks in either basal medium or medium containing VEGF. Cells were examined for gene and protein expression as well as the ability to internalize acetylated low density lipoprotein. With either treatment, endothelial differentiation occurred as evidenced by upregulation of gene and protein expression of typical endothelial markers and the ability to internalize acetylated low density lipoproteins. Interestingly, the addition of VEGF at typical or high concentrations (50 or 100 ng/ml) did not result in differences in gene or protein expression levels of many typical endothelial markers. However, high concentrations of VEGF did significantly increase protein expression of the arterial marker Ephrin‐B1. Thus, VEGF did not accelerate or enhance differentiation of human MSCs towards endothelial cells but was vital for specification of arterial fate. J. Cell. Physiol. 229: 90–96, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24421   open full text
• Circulating miR‐22, miR‐24 and miR‐34a as Novel Predictive Biomarkers to Pemetrexed‐Based Chemotherapy in Advanced Non‐Small Cell Lung Cancer.
  Tindara Franchina, Valeria Amodeo, Giuseppe Bronte, Giuseppina Savio, Giuseppina R.R. Ricciardi, Maria Picciotto, Antonio Russo, Antonio Giordano, Vincenzo Adamo.
  Journal of Cellular Physiology. September 23, 2013
  Pemetrexed has been widely used in patients with advanced non‐small cell lung cancer (NSCLC). The clinical relevance of polymorphisms of folate pathway genes for pemetrexed metabolism have not been fully elucidated yet. The aim of this study was to evaluate the expression levels of circulating miR‐22, miR‐24, and miR‐34a, possibly involved in folate pathway, in NSCLC patients treated with pemetrexed compared with healthy controls and to investigate their impact on patient clinical outcomes. A total of 22 consecutive patients with advanced NSCLC, treated with pemetrexed‐based chemotherapy and 27 age and sex matched healthy controls were included in this preliminary analysis. miR‐22, miR‐24, and miR‐34a targets were identified by TargetScan 6.2 algorithm, validating the involvement of these microRNAs in folate pathway. MicroRNAs were isolated from whole blood and extracted with miRNAeasy Mini Kit (Qiagen). miRNA profiling was performed using Real‐Time PCR. SPSS 17 was used to data analysis. miR‐22, miR‐24, and miR‐34a were found upregulated (P < 0.05) in NSCLC patients versus healthy controls. Higher expression levels were recorded for miR‐34a. Nevertheless, significantly higher miR‐22 expression was observed in patients developing progressive disease (P = 0.03). No significant associations with clinical outcome were recorded for miR‐24 and miR‐34a. Albeit preliminary, these data support the involvement of miR‐22, miR‐24, and miR‐34a in advanced NSCLC. The correlation between high expression of miR‐22 in whole blood and the lack of response in pemetrexed treated NSCLC patients indicates that miR‐22 could represent a novel predictive biomarker for pemetrexed‐based treatment. J. Cell. Physiol. 229: 97–99, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24422   open full text
• Carbon Ion Beams Induce Hepatoma Cell Death by NADPH Oxidase‐Mediated Mitochondrial Damage.
  Chao Sun, Zhenhua Wang, Yang Liu, Yuanyuan Liu, Hongyan Li, Cuixia Di, Zhenhua Wu, Lu Gan, Hong Zhang.
  Journal of Cellular Physiology. September 23, 2013
  Mitochondria are a major source of reactive oxygen species (ROS) and are also the target of cellular ROS. ROS damage to mitochondria leads to dysfunction that further enhances the production of mitochondrial ROS. This feed‐forward vicious cycle between mitochondria and ROS induces cell death. Within a few minutes of radiation exposure, NADPH oxidase is activated to elevate the ROS level. Activated NADPH oxidase might induce the feed‐forward cycle of mitochondria and this is a possible mechanism for cancer cell death induced by heavy ion irradiation. We found that after 4 Gy of 12C6+ ion radiation of HepG2 cells, the NADPH oxidase membrane subunit gp91phox was not involved in enzyme activation through increased expression; however, the subunit p47phox was involved in activation by being translocated to the membrane. 12C6+ ion radiation clearly decreased the ΔΨm of HepG2 cells, increasing mitochondrial DNA damage and inducing cell death. Pretreatment with apocynin (APO, an NADPH oxidase inhibitor) effectively prevented the ΔΨm decrease, mitochondrial DNA damage, and cell death induced by radiation. However, these protective effects were not observed with APO treatment after irradiation exposure. These data demonstrated that NADPH oxidase activation was an initiator in mitochondrial damage. Once mitochondria entered the feed‐forward cycle, cell fate was no longer controlled by NADPH oxidase. Only antioxidants that targeted mitochondria such as MitoQ could break the cycle and release cells from death. J. Cell. Physiol. 229: 100–107, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24424   open full text
• Activated ERK/FOXM1 Pathway by Low‐Power Laser Irradiation Inhibits UVB‐Induced Senescence Through Down‐Regulating p21 Expression.
  Qingzhou Ling, Chengbo Meng, Qun Chen, Da Xing.
  Journal of Cellular Physiology. September 23, 2013
  Cellular senescence is a growth‐arrest program that limits cell proliferation. Low‐power laser irradiation (LPLI) has been demonstrated to promote cell proliferation. However, whether LPLI can inhibit cellular senescence remains unknown. In the present study, to investigate the functional role of LPLI against skin aging, we used ultraviolet radiation b (UVB) to induce cell senescence. We first report that LPLI can delay UVB‐induced cell senescence. The senescence‐associated β‐galactosidase (SA‐β‐Gal) activity and p21 expression, hallmarks of senescent cells, were decreased in the Forkhead box transcription factor FOXM1‐dependent manner under treatment with LPLI. The effect of LPLI was further enhanced with an overexpression of FOXM1, and abolished when FOXM1 was knockdown with short hairpin RNA (shRNA). Furthermore, LPLI activated the extracellular regulated protein kinases (ERK) that was upstream of FOXM1. This led to FOXM1 phosphorylation and nuclear translocation. Nuclear translocation enhanced FOXM1 transcriptional activity and promoted its downstream target gene c‐Myc expression that could inhibit p21 expression. These findings highlight the protective effects of ERK/FOXM1 pathway against UVB‐induced cell senescence, suggesting a potential protecting strategy for treating skin aging by LPLI. J. Cell. Physiol. 229: 108–116, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24425   open full text
• Excess Nitric Oxide Impairs LXR(α)‐ABCA1‐Dependent Cholesterol Efflux in Macrophage Foam Cells.
  Jin‐Feng Zhao, Song‐Kun Shyue, Shing‐Jong Lin, Jeng Wei, Tzong‐Shyuan Lee.
  Journal of Cellular Physiology. September 23, 2013
  Excess nitric oxide (NO) promotes the progression of atherosclerosis by increasing the oxidation of low‐density lipoprotein (LDL) and inflammatory responses. However, little is known about the impact of NO and its underlying molecular mechanism on lipid metabolism of macrophage foam cells. In this study, Oil‐red O staining, cholesterol and triglyceride assay, Dil‐oxidized LDL (oxLDL) binding assay, cholesterol efflux assay, real‐time RT‐PCR and Western blot analysis were used for in vitro experiments. Apolipoprotein E‐deficient (apoE−/−) and apoE and inducible nitric oxide synthase‐deficient (apoE−/−iNOS−/−) mice were as our in vivo models. Treatment with S‐nitroso‐N‐acetyl‐D,L‐penicillamine (SNAP), an NO donor, exacerbated oxLDL‐induced cholesterol accumulation in macrophages, because of reduced efficacy of cholesterol efflux. In addition, SNAP decreased the protein level of ATP‐binding cassette transporter A1 (ABCA1) without affecting scavenger receptor type A (SR‐A), CD36, ABCG1, or SR‐B1 levels. This SNAP‐mediated downregulation of ABCA1 was mainly through the effect of NO but not peroxynitrite. Furthermore, the SNAP‐downregulated ABCA1 was due to the decrease in the liver X receptor α (LXRα)‐dependent transcriptional regulation. Moreover, genetic deletion of iNOS increased the serum capacity of reverse cholesterol efflux and protein expression of LXRα, ABCA1, and SR‐BI in aortas and retarded atherosclerosis in apoE−/− mice. Our findings provide new insights in the pro‐atherogenic effect of excess NO on cholesterol metabolism in macrophages. J. Cell. Physiol. 229: 117–125, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24429   open full text
• Enhanced Prostate Cancer Gene Transfer and Therapy Using a Novel Serotype Chimera Cancer Terminator Virus (Ad.5/3‐CTV).
  Belal M. Azab, Rupesh Dash, Swadesh K. Das, Sujit K. Bhutia, Siddik Sarkar, Xue‐Ning Shen, Bridget A. Quinn, Paul Dent, Igor P. Dmitriev, Xiang‐Yang Wang, David T. Curiel, Maurizio Pellecchia, John C. Reed, Devanand Sarkar, Paul B. Fisher.
  Journal of Cellular Physiology. September 23, 2013
  Few options are available for treating patients with advanced prostate cancer (PC). As PC is a slow growing disease and accessible by ultrasound, gene therapy could provide a viable option for this neoplasm. Conditionally replication‐competent adenoviruses (CRCAs) represent potentially useful reagents for treating PC. We previously constructed a CRCA, cancer terminator virus (CTV), which showed efficacy both in vitro and in vivo for PC. The CTV was generated on a serotype 5‐background (Ad.5‐CTV) with infectivity depending on Coxsackie‐Adenovirus Receptors (CARs). CARs are frequently reduced in many tumor types, including PCs thereby limiting effective Ad‐mediated therapy. Using serotype chimerism, a novel CTV (Ad.5/3‐CTV) was created by replacing the Ad.5 fiber knob with the Ad.3 fiber knob thereby facilitating infection in a CAR‐independent manner. We evaluated Ad.5/3‐CTV in comparison with Ad.5‐CTV in low CAR human PC cells, demonstrating higher efficiency in inhibiting cell viability in vitro. Moreover, Ad.5/3‐CTV potently suppressed in vivo tumor growth in a nude mouse xenograft model and in a spontaneously induced PC that develops in Hi‐myc transgenic mice. Considering the significant responses in a Phase I clinical trial of a non‐replicating Ad.5‐mda‐7 in advanced cancers, Ad.5/3‐CTV may exert improved therapeutic benefit in a clinical setting. J. Cell. Physiol. 229: 34–43, 2014. © 2013 Wiley Periodicals, Inc.

  September 23, 2013   doi: 10.1002/jcp.24408   open full text
• Characterization, isolation and culture of mouse and human spermatogonial stem cells.
  Ying Guo, Yanan Hai, Yuehua Gong, Zheng Li, Zuping He.
  Journal of Cellular Physiology. September 20, 2013
  Spermatogenesis is a special process by which spermatogonial stem cells (SSCs) divide and differentiate to male gametes called mature spermatozoa. SSCs are the unique cells because they are adult stem cells that transmit genetic information to subsequent generations. Accumulating evidence has demonstrated that SSCs can be reprogrammed to acquire pluripotency to become embryonic stem‐like cells that differentiate into all cell lineages of the three germ layers, highlighting potential important applications of SSCs for regenerative medicine. Recent studies from peers and us have made great achievements on the characterization, isolation and culture of mouse and human SSCs, which could lead to better understanding the biology of SSCs and the applications of SSCs in both reproductive and regenerative medicine. In this review, we first compared the cell identity and biochemical phenotypes between mouse SSCs and human SSCs. Notably, the cell types of mouse and human SSCs are distinct, and human SSCs share some but not all phenotypes with mouse SSCs. The approaches for isolating SSCs as well as short‐ and long‐ term culture of mouse SSCs and short‐period culture of human SSCs were also discussed. We further addressed the new advances on the self‐renewal of SSCs with an aim to establish the long‐term culture of human SSCs which has not yet been achieved. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 20, 2013   doi: 10.1002/jcp.24471   open full text
• Identification of pivotal cellular factors involved in HPV‐induced dysplastic and neoplastic cervical pathologies.
  Stefano Mattarocci, Claudia Abbruzzese, Anna M. Mileo, Mariantonia Carosi, Edoardo Pescarmona, Carmen Vico, Antonio Federico, Enrico Vizza, Giacomo Corrado, Ivan Arisi, Armando Felsani, Marco G. Paggi.
  Journal of Cellular Physiology. September 16, 2013
  Cervical carcinoma represents the paradigm of virus‐induced cancers, where virtually all cervical cancers come from previous “high‐risk” HPV infection. The persistent expression of the HPV viral oncoproteins E6 and E7 is responsible for the reprogramming of fundamental cellular functions in the host cell, thus generating a noticeable, yet only partially explored, imbalance in protein molecular networks and cell signaling pathways. Eighty‐eight cellular factors, identified as HPV direct or surrogate targets, were chosen and monitored in a retrospective analysis for their mRNA expression in HPV‐induced cervical lesions, from dysplasia to cancer. Real‐time quantitative PCR (qPCR) was performed by using formalin‐fixed, paraffin embedded archival samples. Gene expression analysis identified 40 genes significantly modulated in LSIL, HSIL and squamous cervical carcinoma. Interestingly, among these, the expression level of a panel of four genes, TOP2A, CTNNB1, PFKM and GSN, was able to distinguish between normal tissues and cervical carcinomas. Immunohistochemistry was also done to assess protein expression of two genes among those up‐regulated during the transition between dysplasia and carcinoma, namely E2F1 and CDC25A, and their correlation with clinical parameters. Besides the possibility of significantly enhancing the use of some of these factors in diagnostic or prognostic procedures, these data clearly outline specific pathways, and thus key biological processes, altered in cervical dysplasia and carcinoma. Deeper insight on how these molecular mechanisms work may help widen the spectrum of novel innovative approaches to these virus‐induced cell pathologies. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 16, 2013   doi: 10.1002/jcp.24465   open full text
• In vitro mimics of bone remodeling and the vicious cycle of cancer in bone.
  Venkatesh Krishnan, Erwin A. Vogler, Donna M. Sosnoski, Andrea M. Mastro.
  Journal of Cellular Physiology. September 11, 2013
  Bone remodeling is a natural process that enables growth and maintenance of the skeleton. It involves the deposition of mineralized matrix by osteoblasts and resorption by osteoclasts. Several cancers that metastasize to bone negatively perturb the remodeling process through a series of interactions with osteoclasts, and osteoblasts. These interactions have been described as the “vicious cycle” of cancer metastasis in bone. Due to the inaccessibility of the skeletal tissue it is difficult to study this system in vivo. In contrast, standard tissue culture lacks sufficient complexity. We have developed a specialized three‐dimensional culture system that permits growth of a non‐vascularized, multiple‐cell‐layer of mineralized osteoblastic tissue from pre‐osteoblasts. In this study, the essential properties of bone remodeling were created in vitro by co‐culturing the mineralized collagenous osteoblastic tissue with actively resorbing osteoclasts followed by reinfusion with proliferating pre‐osteoblasts. Cell‐cell and cell‐matrix interactions were determined by confocal microscopy as well as by assays for cell specific cytokines and growth factors. Osteoclasts, differentiated in the presence of osteoblasts, led to degradation of the collagen‐rich extracellular matrix. Further addition of metastatic breast cancer cells to the co‐culture mimicked the vicious cycle; i.e. there was a further reduction in osteoblastic tissue thickness, an increase in osteoclastogenesis, chemotaxis of cancer cells to osteoclasts and formation of cancer cells into large colonies. The resulting model system permits detailed study of fundamental osteobiological and osteopathological processes in a manner that will enhance development of therapeutic interventions to skeletal diseases. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 11, 2013   doi: 10.1002/jcp.24464   open full text
• Pin1‐mediated prolyl isomerization of Runx1 affects PU.1 expression in pre‐monocytes.
  Rabia Islam, Won‐Joon Yoon, Kyung‐Mi Woo, Jeong‐Hwa Baek, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. September 03, 2013
  Regulation of the hematopoietic transcription factor PU.1, a member of the ETS family, plays a critical role in the development of blood cells and in leukemia. The dosage of PU.1 has been shown to cause a shift in myelomonocytic progenitor fate. Pin1 is a unique substrate‐specific enzyme that can isomerize phospho‐Ser/Thr‐Pro peptide bonds, accelerating the conformational change in its substrates between a cis and a trans form. Such activity has been demonstrated to be a tightly controlled mechanism regulating a wide variety of protein functions under both normal physiological and pathological conditions. We have previously reported that a conformational change in Runx2 induced by Pin1 is essential for its function in osteogenesis in vitro and in vivo. In this study, we show that the Pin1‐mediated conformational change in Runx1 enhances its acetylation and stabilization and, consequently, enhances its transacting activity. The increased acetylation of Runx1 represses PU.1 transcription in pre‐monocytes. Conversely, the lack of (or the inhibition of) Pin1 increases PU.1 transcription in vitro and in vivo in pre‐monocytes and in the spleen tissue. Pin1 KO mice have an increased CD11b+/F4/80+ cell population and F4/80 protein expression in spleen. From our data, we can conclude that the conformational change in Runx1 induced by Pin1 represses PU.1 transcription in pre‐monocytes and influences the commitment to the monocyte lineage. The dosage of PU.1 is a crucial factor in AML (Acute myeloid leukemia), and Pin1 may thus be a useful target for controlling PU.1‐dependent hematopoiesis, as well as leukemogenesis. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 03, 2013   doi: 10.1002/jcp.24462   open full text
• Angiotensin II activation of canonical transient receptor potential‐6 (TRPC6) channels in rat podocytes requires generation of reactive oxygen species.
  Marc Anderson, Hila Roshanravan, Justin Khine, Stuart E. Dryer.
  Journal of Cellular Physiology. September 03, 2013
  Angiotensin II (AII) plays a major role in the progression of chronic kidney diseases. Podocytes are essential components of the ultrafiltration apparatus, and are targets for AII signaling. AII has been shown to increase generation of reactive oxygen species (ROS) in podocytes. Canonical transient receptor potential‐6 (TRPC6) channels stimulate Ca2+ influx in podocytes, and have been implicated in glomerular disease. We observed that AII increased cationic currents in rat podocytes in an isolated glomerulus preparation in which podocytes are still attached to the underlying capillary. This effect was completely blocked by SKF‐96365, by micromolar La3+, and by siRNA knockdown of TRPC6, indicating that TRPC6 is the primary source of Ca2+ influx mobilized by endogenously expressed angiotensin II receptors in these cells. These responses were also blocked by the AT1R antagonist losartan, the phospholipase C inhibitor D‐609, and by inhibition of G protein signaling. The pan‐protein kinase C inhibitor chelerythrine had no effect. Importantly, pretreating podocytes with the ROS quencher manganese (III) tetrakis (4‐benzoic acid) porphyrin chloride (MnTBAP) eliminated AII activation of TRPC6. Significant reductions of AII effects on podocyte TRPC6 were also observed after pre‐treatment with NADPH oxidase inhibitors apocynin or diphenylene iodonium (DPI). These data suggest that ROS production permits activation of TRPC6 channels by G protein and PLC‐dependent cascades initiated by AII acting on AT1Rs in podocytes. This pathway also provides a basis whereby two forms of cellular stress–oxidative stress and Ca2+ overload–converge on common pathways relevant to disease. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 03, 2013   doi: 10.1002/jcp.24461   open full text
• Wnt16 is involved in intramembranous ossification and suppresses osteoblast differentiation through the Wnt/β‐catenin pathway.
  Zheng Jiang, Johannes W. Von den Hoff, Ruurd Torensma, Liuyan Meng, Zhuan Bian.
  Journal of Cellular Physiology. September 03, 2013
  In the course of embryonic development skeletal elements form either through intramembranous or endochondral ossification. Wnt proteins play diverse roles during vertebrate skeletal development. Wnt16 is a key factor in developing long bones, but its exact role in craniofacial bone formation remains unclear. This study was initially undertaken to investigate the expression of Wnt16 during craniofacial bone development in mouse embryos. Wnt16 expression in the osteoid of calvaria, maxilla, and mandible started later than that of ALP and osteocalcin, but before mineralization of the craniofacial bones, suggesting that Wnt16 is involved in intramembranous ossification in the head. To confirm this, MC3T3‐E1 cells were transfected with an adenovirus containing Wnt16 (Ad‐Wnt16). Ad‐Wnt16 cells showed decreased ALP activity and less mineralized nodule formations compared with control cells. In addition, the mRNA levels of osteogenic markers were reduced. Moreover, Wnt16 activated β‐catenin signaling in MC3T3‐E1 cells at both transcription and protein levels as shown by a TOPflash luciferase reporter gene assay and western blot analysis. On the other hand, Wnt/β‐catenin pathway blockade by Dickkopf 1 abrogated the suppression of mineralization by Wnt16. Our findings suggest that Wnt16 is involved in intramembranous ossification and suppresses osteoblast differentiation through the Wnt/β‐catenin pathway. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 03, 2013   doi: 10.1002/jcp.24460   open full text
• Potential role of voltage‐sensing phosphatases in regulation of cell structure through the production of PI(3,4)P2.
  Shinji Yamaguchi, Tatsuki Kurokawa, Ikuko Taira, Naoya Aoki, Souhei Sakata, Yasushi Okamura, Koichi J. Homma.
  Journal of Cellular Physiology. September 02, 2013
  Voltage‐sensing phosphatase, VSP, consists of the transmembrane domain, operating as the voltage sensor, and the cytoplasmic domain with phosphoinositide‐phosphatase activities. The voltage sensor tightly couples with the cytoplasmic phosphatase and membrane depolarization induces dephosphorylation of several species of phosphoinositides. VSP gene is conserved from urochordate to human. There are some diversities among VSP ortholog proteins; range of voltage of voltage sensor motions as well as substrate selectivity. In contrast with recent understandings of biophysical mechanisms of VSPs, little is known about its physiological roles. Here we report that chick ortholog of VSP (designated as Gg‐VSP) induces morphological feature of cell process outgrowths with round cell body in DF‐1 fibroblasts upon its forced expression. Expression of the voltage sensor mutant, Gg‐VSPR153Q with shifted voltage dependence to a lower voltage led to more frequent changes of cell morphology than the wild‐type protein. Coexpression of PTEN that dephosphorylates PI(3,4)P2 suppressed this effect by Gg‐VSP, indicating that the increase of PI(3,4)P2 leads to changes of cell shape. In addition, visualization of PI(3,4)P2 with the fluorescent protein fused with the TAPP1‐derived pleckstrin homology (PH) domain suggested that Gg‐VSP influenced the distribution of PI(3,4)P2. These findings raise a possibility that one of the VSP's functions could be to regulate cell morphology through voltage‐sensitive tuning of phosphoinositide profile. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  September 02, 2013   doi: 10.1002/jcp.24463   open full text
• Morphological effects on expression of growth differentiation factor 15 (GDF15), a marker of metastasis.
  Koh Meng Aw Yong, Yu Zeng, Donald Vindivich, Jude M. Phillip, Pei‐Hsun Wu, Denis Wirtz, Robert H. Getzenberg.
  Journal of Cellular Physiology. August 31, 2013
  Cancer cells typically demonstrate altered morphology during the various stages of disease progression as well as metastasis. While much is known about how altered cell morphology in cancer is a result of genetic regulation, less is known about how changes in cell morphology affect cell function by influencing gene expression. In this study, we altered cell morphology in different types of cancer cells by disrupting the actin cytoskeleton or by modulating attachment and observed a rapid up‐regulation of growth differentiation factor 15 (GDF15), a member of the transforming growth factor‐beta (TGF‐β) super‐family. Strikingly, this up‐regulation was sustained as long as the cell morphology remained altered but was reversed upon allowing cell morphology to return to its typical configuration. The potential significance of these findings was examined in vivo using a mouse model: a small number of cancer cells grown in diffusion chambers that altered morphology increased mouse serum GDF15. Taken together, we propose that during the process of metastasis, cancer cells experience changes in cell morphology, resulting in the increased production and secretion of GDF15 into the surrounding environment. This indicates a possible relationship between serum GDF15 levels and circulating tumor cells may exist. Further investigation into the exact nature of this relationship is warranted. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 31, 2013   doi: 10.1002/jcp.24458   open full text
• Calnexin silencing in mouse neonatal cardiomyocytes induces Ca2+ cycling defects, ER stress, and apoptosis.
  Nicolas Bousette, Cynthia Abbasi, Roxana Chis, Anthony O. Gramolini.
  Journal of Cellular Physiology. August 29, 2013
  Calnexin (CNX) is an endoplasmic reticulum (ER) quality control chaperone that has been implicated in ER stress. ER stress is a prominent pathological feature of various pathologic conditions, including cardiovascular diseases. However, the role of CNX and ER stress has not been studied in the heart. In the present study, we aimed to characterize the role of CNX in cardiomyocyte physiology with respect to ER stress, apoptosis, and cardiomyocyte Ca2+ cycling. We demonstrated significantly decreased CNX mRNA and protein levels by LentiVector mediated transduction of targeting shRNAs. CNX silenced cardiomyocytes exhibited ER stress as evidenced by increased GRP78 and ATF6 protein levels, increased levels of spliced XBP1 mRNA, ASK‐1, ERO1a, and CHOP mRNA levels. CNX silencing also led to significant activation of caspases‐3 and ‐9. This activation of caspases was associated with hallmark morphological features of apoptosis including loss of sarcomeric organization and nuclear integrity. Ca2+ imaging in live cells showed that CNX silencing resulted in Ca2+ transients with significantly larger amplitudes but decreased frequency and Ca2+ uptake rates in the basal state. Interestingly, 5mM caffeine stimulated Ca2+ transients were similar between control and CNX silenced cardiomyocytes. Finally, we demonstrated that CNX silencing induced the expression of the L‐type voltage dependent calcium channel (CAV1.2) but reduced the expression of the sarcoplasmic reticulum ATPase (SERCA2a). In conclusion, this is the first study to demonstrate CNX has a specific role in cardiomyocyte viability and Ca2+ cycling through its effects on ER stress, apoptosis and Ca2+ channel expression. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 29, 2013   doi: 10.1002/jcp.24459   open full text
• MicroRNA‐302a sensitizes testicular embryonal carcinoma cells to cisplatin‐induced cell death.
  Lin Liu, Jie Lian, Huijuan Zhang, Hui Tian, Meng Liang, Mianmian Yin, Fei Sun.
  Journal of Cellular Physiology. August 23, 2013
  Cisplatin is a commonly used chemotherapeutic agent for the treatment of several human malignancies, such as testicular germ cell tumors (TGCT). The toxic effects persist and those that are present long after chemotherapy affect the overall quality of life of patients. MicroRNAs (miRNAs) play important roles in the responses of cancer cells to chemotherapy and have been shown to modulate cell sensitivity to chemotherapeutic drugs. However, the relationship between miRNA expression and cisplatin sensitivity of TGCT has not been fully explored. In this study, the effects of miR‐302a on cisplatin cytotoxicity in TGCT‐derived cell line NTERA‐2 (NT2) were evaluated. We found that expression levels of miR‐302a were increased in cisplatin‐treated NT2 cells. Up‐regulation of miR‐302a significantly increased the sensitivity of NT2 cells to cisplatin by enhancing cisplatin‐induced G2/M phase arrest and the subsequent progression to apoptosis. MiR‐302a also increased the killing effects of cisplatin by lowering the apoptotic threshold; the same result was also observed in another TGCT‐derived cell line, NCCIT. Furthermore, miR‐302a‐enhanced cisplatin sensitivity was partially mediated through the down‐regulation of p21 in NT2 cells. MiR‐302a induced apoptosis was further enhanced by silencing of p53 in NT2 cells. p53 levels were inversely associated with the expression of Oct4, Sox2, and Nanog in response to cisplatin. Thus, targeting miR‐302a may offer new therapeutic interventions in TGCT. J. Cell. Physiol. 228: 2294–2304, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24394   open full text
• Some mechanisms of FLIP expression in inhibition of HIV‐1 replication in Jurkat cells, CD4+ T cells and PBMCs.
  Jiying Tan, Xue Wang, Krishnakumar Devadas, Jiangqin Zhao, Panhe Zhang, Indira Hewlett.
  Journal of Cellular Physiology. August 23, 2013
  HIV‐1 infection and replication are affected by host factors. Recent studies demonstrate that molecules from apoptotic pathways regulate HIV‐1 replication. Therefore, studies on effects of host factors that maintain host cell survival and influence HIV‐1 replication are critical to understanding the mechanisms of HIV‐1 replicative cycle. Using the susceptible Jurkat cell line, CD4+ T cells, and peripheral blood mononuclear cells (PBMCs), we studied the role of FLIP, an inhibitor of caspase‐8, in HIV‐1 production. Full length cellular FLIP (cFLIP) inhibited HIV‐1 replication in these cells. cFLIP upregulated the expression of viral restriction factors, such as TRIM5, Apobec3G, and Bst2/tetherin, decreased nuclear factor 1C expression and inactivated ERK and p38 induced by HIV‐1 in Jurkat cells. cFLIP blocked the trafficking of gp120 and Gag p24 capsid protein into lipid rafts with inhibition of Tsg101 and Alix in ESCRT signaling pathway. cFLIP also promoted Bst2/tetherin trafficking into lipid rafts. These results indicate that cFLIP may inhibit the HIV‐1 replication cycle at multiple steps, including viral RNA release, transcription, traffic and assembly. We also found that cFLIP expression downregulated Fas expression and inactivated FADD in the Fas‐mediated apoptotic pathway. The inactivated FADD also inhibited HIV‐1 replication. J. Cell. Physiol. 228: 2305–2313, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24397   open full text
• miR‐10a restores human mesenchymal stem cell differentiation by repressing KLF4.
  Jiao Li, Jun Dong, Zhen‐hui Zhang, Dong‐Cheng Zhang, Xiang‐Yu You, Yun Zhong, Min‐Sheng Chen, Shi‐Ming Liu.
  Journal of Cellular Physiology. August 23, 2013
  miRNAs have recently been shown to play a significant role in human aging. However, data demonstrating the effects of aging‐related miRNAs in human mesenchymal stem cells (hMSCs) are limited. We observed that hMSC differentiation decreased with aging. We also identified that miR‐10a expression was significantly decreased with age by comparing the miRNA expression of hMSCs derived from young and aged individuals. Therefore, we hypothesized that the downregulation of miR‐10a may be associated with the decreased differentiation capability of hMSCs from aged individuals. Lentiviral constructs were used to up‐ or downregulate miR‐10a in young and old hMSCs. Upregulation of miR‐10a resulted in increased differentiation to adipogenic, osteogenic, and chondrogenic lineages and in reduced cell senescence. Conversely, downregulation of miR‐10a resulted in decreased cell differentiation and increased cell senescence. A chimeric luciferase reporter system was generated, tagged with the full‐length 3′‐UTR region of KLF4 harboring the seed‐matched sequence with or without four nucleotide mutations. These constructs were cotransfected with the miR‐10a mimic into cells. The luciferase activity was significantly repressed by the miR‐10a mimic, proving the direct binding of miR‐10a to the 3′‐UTR of KLF4. Direct suppression of KLF4 in aged hMSCs increased cell differentiation and decreased cell senescence. In conclusion, miR‐10a restores the differentiation capability of aged hMSCs through repression of KLF4. Aging‐related miRNAs may have broad applications in the restoration of cell dysfunction caused by aging. J. Cell. Physiol. 228: 2324–2336, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24402   open full text
• Sirtuins' modulation of autophagy.
  Fanny Ng, Bor Luen Tang.
  Journal of Cellular Physiology. August 23, 2013
  The sirtuin family of class III histone deacetylases has been extensively implicated in modulating a myriad of cellular processes, including energy metabolism, stress response, cell/tissue survival and malignancy. Recent studies have also identified multifaceted roles for Sirt1 and Sirt2 in the regulation of autophagy. Sirt1 could influence autophagy directly via its deacetylation of key components of the autophagy induction network, such as the products of autophagy genes (Atg) 5, 7, and 8. Nucleus‐localized Sirt1 is also known to induce the expression of autophagy pathway components through the activation of FoxO transcription factor family members. The perception of a linear Sirt1‐FoxO axis in autophagy induction is complicated by recent findings that acetylated FoxO1 could bind to Atg7 in the cytoplasm and affect autophagy directly. This occurs with prolonged stress signaling, with FoxO1's continuous dissociation from cytoplasmic Sirt2 and its consequential hyperacetylation. FoxO‐mediated nuclear transcription may induce/enhance autophagy in ways that are different compared to cytoplasmic FoxO, thereby leading to contrasting (cell survival versus cell death) outcomes. FoxO and Sirt1 are both subjected to regulation by stress signaling (e.g., through the c‐Jun N‐terminal kinases (JNK)) in the context of autophagy induction, which are also critical in determining between cell survival and death in a context‐dependent manner. We discussed here the emerging molecular intricacies of sirtuins' connections with autophagy. A good understanding of these connections would serve to consolidate a framework of mechanisms underlying Sirt1's protective effects in multiple physiological systems. J. Cell. Physiol. 228: 2262–2270, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24399   open full text
• Expression of microRNA‐184 in keratinocytes represses argonaute 2.
  Julian C. Roberts, Richard B. Warren, Christopher E.M. Griffiths, Kehinde Ross.
  Journal of Cellular Physiology. August 23, 2013
  Interleukin‐22 (IL‐22) is a proinflammatory cytokine that has been associated with the pathogenesis of inflammatory skin disorders. However, the impact of IL‐22 on microRNA (miRNA) expression in epidermal keratinocytes is unknown. Here we show that IL‐22 induces miR‐184 in reconstituted human epidermis (RHE) and in the HaCaT keratinocyte cell line. Exposure to IL‐22 increased miR‐184 expression 8‐ and 15‐fold in RHE and HaCaT cells, respectively. Oncostatin M, an unrelated proinflammatory cytokine, also raised miR‐184 expression in RHE and HaCaT keratinocytes. Pharmacologic and genetic inhibition demonstrated that cytokine‐induced expression of miR‐184 was mediated by signal transducer and activation of transcription 3 (STAT3). Argonaute 2 (AGO2), a member of the RNA‐induced silencing complex (RISC), is a predicted miR‐184 target. Using protein, messenger RNA and reporter analyses, we found that miR‐184 regulates the expression of AGO2. We conclude that cytokine‐induced miR‐184 attenuates AGO2 expression in keratinocytes. J. Cell. Physiol. 228: 2314–2323, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24401   open full text
• Pin1‐mediated Runx2 modification is critical for skeletal development.
  Won‐Joon Yoon, Rabia Islam, Young‐Dan Cho, Kyung‐Mi Woo, Jeong‐Hwa Baek, Takafumi Uchida, Toshihisa Komori, Andre van Wijnen, Janet L. Stein, Jane B. Lian, Gary S. Stein, Je‐Yong Choi, Suk‐Chul Bae, Hyun‐Mo Ryoo.
  Journal of Cellular Physiology. August 23, 2013
  Runx2 is the master transcription factor for bone formation. Haploinsufficiency of RUNX2 is the genetic cause of cleidocranial dysplasia (CCD) that is characterized by hypoplastic clavicles and open fontanels. In this study, we found that Pin1, peptidyl prolyl cis–trans isomerase, is a critical regulator of Runx2 in vivo and in vitro. Pin1 mutant mice developed CCD‐like phenotypes with hypoplastic clavicles and open fontanels as found in the Runx2+/− mice. In addition Runx2 protein level was significantly reduced in Pin1 mutant mice. Moreover Pin1 directly interacts with the Runx2 protein in a phosphorylation‐dependent manner and subsequently stabilizes Runx2 protein. In the absence of Pin1, Runx2 is rapidly degraded by the ubiquitin‐dependent protein degradation pathway. However, Pin1 overexpression strongly attenuated uniquitin‐dependent Runx2 degradation. Collectively conformational change of Runx2 by Pin1 is essential for its protein stability and possibly enhances the level of active Runx2 in vivo. J. Cell. Physiol. 228: 2377–2385, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24403   open full text
• SWI/SNF chromatin remodeling enzymes are associated with cardiac hypertrophy in a genetic rat model of hypertension.
  Aanchal Mehrotra, Bina Joe, Ivana L. de la Serna.
  Journal of Cellular Physiology. August 23, 2013
  Pathological cardiac hypertrophy is characterized by a sustained increase in cardiomyocyte size and re‐activation of the fetal cardiac gene program. Previous studies implicated SWI/SNF chromatin remodeling enzymes as regulators of the fetal cardiac gene program in surgical models of cardiac hypertrophy. Although hypertension is a common risk factor for developing cardiac hypertrophy, there has not yet been any investigation into the role of SWI/SNF enzymes in cardiac hypertrophy using genetic models of hypertension. In this study, we tested the hypothesis that components of the SWI/SNF complex are activated and recruited to promoters that regulate the fetal cardiac gene program in hearts that become hypertrophic as a result of salt induced hypertension. Utilizing the Dahl salt‐sensitive (S) rat model, we found that the protein levels of several SWI/SNF subunits required for heart development, Brg1, Baf180, and Baf60c, are elevated in hypertrophic hearts from S rats fed a high salt diet compared with normotensive hearts from Dahl salt‐resistant (R) rats fed the same diet. Furthermore, we detected significantly higher levels of SWI/SNF subunit enrichment as well as evidence of more accessible chromatin structure on two fetal cardiac gene promoters in hearts from S rats compared with R rats. Our data implicate SWI/SNF chromatin remodeling enzymes as regulators of gene expression in cardiac hypertrophy resulting from salt induced hypertension. Thus we provide novel insights into the epigenetic mechanisms by which salt induced hypertension leads to cardiac hypertrophy. J. Cell. Physiol. 228: 2337–2342, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24404   open full text
• Gene methylation in rectal cancer: Predictive marker of response to chemoradiotherapy?
  Chiara Molinari, Valentina Casadio, Flavia Foca, Chiara Zingaretti, Massimo Giannini, Andrea Avanzolini, Enrico Lucci, Luca Saragoni, Alessandro Passardi, Dino Amadori, Daniele Calistri, Wainer Zoli.
  Journal of Cellular Physiology. August 23, 2013
  Although numerous studies have focused on the link between CpG island methylator phenotypes and the development of colorectal cancer, few studies have dealt specifically with methylation profiling in rectal cancer and its role in predicting response to neoadjuvant chemoradiotherapy (NCRT). We characterized methylation profiles in normal and neoplastic tissue samples from patients with rectal cancer and assessed the role of this molecular profile in predicting chemoradioactivity. We evaluated 74 pretreatment tumor samples and 16 apparently normal tissue biopsies from rectal cancer patients submitted to NCRT. The methylation profile of 24 different tumor suppressor genes was analyzed from FFPE samples by methylation‐specific multiplex ligation‐dependent probe amplification (MS‐MLPA). Methylation status was studied in relation to tissue type and clinical pathological parameters, in particular, pathological response evaluated by tumor regression grade (TRG). ESR1, CDH13, RARB, IGSF4, and APC genes showed high methylation levels in tumor samples (range 18.92–49.77) with respect to normal tissue. Methylation levels of the remaining genes were low and similar in both normal (range 1.91–14.56) and tumor tissue (range 1.84–11). Analysis of the association between methylation and response to therapy in tumor samples showed that only TIMP3 methylation status differed significantly within the four TRG classes (ANOVA, P < 0.05). Results from the present explorative study suggest that quantitative epigenetic classification of rectal cancer by MS‐MLPA clearly distinguishes tumor tissue from apparently normal mucosa. Conversely, with the exception of TIMP3 gene, the methylation of selected genes does not seem to correlate with response to NCRT. J. Cell. Physiol. 228: 2343–2349, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24405   open full text
• Differential effect of hypoxia on etoposide‐induced DNA damage response and p53 regulation in different cell types.
  Audrey Sermeus, Magali Rebucci, Maude Fransolet, Lionel Flamant, Déborah Desmet, Edouard Delaive, Thierry Arnould, Carine Michiels.
  Journal of Cellular Physiology. August 23, 2013
  Among the main causes of cancer cell resistance to chemotherapy are p53 mutation and hypoxic tumor microenvironment. However, the effect of hypoxia can be very different from one cell type to the other. We studied the effect of hypoxia on the etoposide‐induced cell death in two cancer cell lines, HepG2 and A549 cells. Hypoxia decreased etoposide‐induced apoptosis in HepG2 cells but not in A549 cells. Here, we evidenced two pathways, known to play important roles in cancer cell resistance, that are differently affected by hypoxia in these two cell types. First, in HepG2 cells, hypoxia decreased p53 protein level and activity by acting post‐transcriptionally and independently of HIF‐1. The results suggest an effect of hypoxia on p53 translation. On the other hand, in A549 cells, no effect of hypoxia was observed on p53 level. Secondly, hypoxia decreased DNA damage response in HepG2 cells while this was not the case in A549 cells. Indeed, a decrease in the phosphorylation level of CHK2 and H2AX with a decrease in ATM activity was observed. Importantly, these results evidenced that hypoxia can prevent cancer cell apoptosis by acting at different levels in the cell and that these effects are strongly cell‐type dependent. J. Cell. Physiol. 228: 2365–2376, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24409   open full text
• Dentistry proteomics: From laboratory development to clinical practice.
  Taia M. B. Rezende, Stella M. F. Lima, Bernardo A. Petriz, Osmar N. Silva, Mirna S. Freire, Octávio L. Franco.
  Journal of Cellular Physiology. August 23, 2013
  Despite all the dental information acquired over centuries and the importance of proteome research, the cross‐link between these two areas only emerged around mid‐nineties. Proteomic tools can help dentistry in the identification of risk factors, early diagnosis, prevention, and systematic control that will promote the evolution of treatment in all dentistry specialties. This review mainly focuses on the evolution of dentistry in different specialties based on proteomic research and how these tools can improve knowledge in dentistry. The subjects covered are an overview of proteomics in dentistry, specific information on different fields in dentistry (dental structure, restorative dentistry, endodontics, periodontics, oral pathology, oral surgery, and orthodontics) and future directions. There are many new proteomic technologies that have never been used in dentistry studies and some dentistry areas that have never been explored by proteomic tools. It is expected that a greater integration of these areas will help to understand what is still unknown in oral health and disease. J. Cell. Physiol. 228: 2271–2284, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24410   open full text
• The Xenopus TRPV6 homolog encodes a Mg2+‐permeant channel that is inhibited by interaction with TRPC1.
  Raphael Courjaret, Satanay Hubrack, Arwa Daalis, Maya Dib, Khaled Machaca.
  Journal of Cellular Physiology. August 23, 2013
  The TRP gene family encodes primarily cation non‐selective, Ca2+ permeant channels that are involved in a dizzying array of sensory mechanisms. Two channels in this large family TRPV5 and TRPV6 are highly Ca2+ selective and are expressed in epithelia where they are important in Ca2+ uptake. TRPV5/6 are constitutively active, yet the mechanisms regulating their activation in native tissue remains elusive. Here we functionally characterize the Xenopus TRPV6 homolog. xTRPV6 is expressed in the oocyte and encodes a channel that is permeant to divalents including Ca2+, and displays a high permeability to Mg2+. The oocyte does not exhibit functional TRPV6‐like current at rest, showing that the endogenous channel is somehow maintained in an inactive state. We show that endogenous as well as overexpressed xTRPV6 interacts with xTRPC1 and that this interaction inhibits xTRPV6 currents. As such TRPC1 is likely to regulate the activity of TRPV6 under physiological conditions. J. Cell. Physiol. 228: 2386–2398, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24411   open full text
• Cbl‐family proteins as regulators of cytoskeleton‐dependent phenomena.
  Hojin Lee, Alexander Y. Tsygankov.
  Journal of Cellular Physiology. August 23, 2013
  Although the role of Cbl‐family proteins as key cellular regulators has been established, phenomena regulated in a Cbl‐dependent fashion are multiple and the mechanisms mediating the effects of Cbl proteins are diverse. This finding makes it important to consider different phenomena affected by functions of Cbl proteins individually. Among effects of Cbl on various biological functions there are many cases of regulation of cellular phenomena related to cytoskeletal rearrangements, such as cell adhesion, motility and invasion. Some of these regulatory functions are mediated by adaptor‐type interactions of Cbl, especially by Cbl‐dependent modulation of phosphatidyl‐inositol‐3′ kinase (PI3K), while others are caused by Cbl‐dependent ubiquitylation of various cytoskeletal and regulatory proteins, identified and unidentified. The role of Cbl in regulation of cytoskeleton‐dependent cellular functions is discussed in this review. J. Cell. Physiol. 228: 2285–2293, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24412   open full text
• Testosterone increases GLUT4‐dependent glucose uptake in cardiomyocytes.
  Carlos Wilson, Ariel Contreras‐Ferrat, Nataly Venegas, César Osorio‐Fuentealba, Mario Pávez, Katherine Montoya, Javier Durán, Rodrigo Maass, Sergio Lavandero, Manuel Estrada.
  Journal of Cellular Physiology. August 23, 2013
  Testosterone exerts important effects in the heart. Cardiomyocytes are target cells for androgens, and testosterone induces rapid effects via Ca2+ release and protein kinase activation and long‐term effects via cardiomyocyte differentiation and hypertrophy. Furthermore, it stimulates metabolic effects such as increasing glucose uptake in different tissues. Cardiomyocytes preferentially consume fatty acids for ATP production, but under particular circumstances, glucose uptake is increased to optimize energy production. We studied the effects of testosterone on glucose uptake in cardiomyocytes. We found that testosterone increased uptake of the fluorescent glucose analog 2‐(N‐(7‐nitrobenz‐2‐oxa‐1, 3‐diazol‐4‐yl)amino)‐2‐deoxyglucose and [3H]2‐deoxyglucose, which was blocked by the glucose transporter 4 (GLUT4) inhibitor indinavir. Testosterone stimulation in the presence of cyproterone or albumin‐bound testosterone‐induced glucose uptake, which suggests an effect that is independent of the intracellular androgen receptor. To determine the degree of GLUT4 cell surface exposure, cardiomyocytes were transfected with the plasmid GLUT4myc‐eGFP. Subsequently, testosterone increased GLUT4myc‐GFP exposure at the plasma membrane. Inhibition of Akt by the Akt‐inhibitor‐VIII had no effect. However, inhibition of Ca2+/calmodulin protein kinase (CaMKII) (KN‐93 and autocamtide‐2 related inhibitory peptide II) and AMP‐activated protein kinase (AMPK) (compound C and siRNA for AMPK) prevented glucose uptake induced by testosterone. Moreover, GLUT4myc‐eGFP exposure at the cell surface caused by testosterone was also abolished after CaMKII and AMPK inhibition. These results suggest that testosterone increases GLUT4‐dependent glucose uptake, which is mediated by CaMKII and AMPK in cultured cardiomyocytes. Glucose uptake could represent a mechanism by which testosterone increases energy production and protein synthesis in cardiomyocytes. J. Cell. Physiol. 228: 2399–2407, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24413   open full text
• Knockdown of core binding factorβ alters sphingolipid metabolism.
  Adam H. Greer, Thomas Yong, Katie Fennell, Yara W. Moustafa, Marcie Fowler, Floyd Galiano, Shu‐Wing Ng, Ross S. Berkowitz, James Cardelli, Shari Meyers, J. Nathan Davis.
  Journal of Cellular Physiology. August 23, 2013
  Core binding factor (CBF) is a heterodimeric transcription factor containing one of three DNA‐binding proteins of the Runt‐related transcription factor family (RUNX1–3) and the non‐DNA‐binding protein, CBFβ. RUNX1 and CBFβ are the most common targets of chromosomal rearrangements in leukemia. CBF has been implicated in other cancer types; for example RUNX1 and RUNX2 are implicated in cancers of epithelial origin, including prostate, breast, and ovarian cancers. In these tumors, CBF is involved in maintaining the malignant phenotype and, when highly over‐expressed, contributes to metastatic growth in bone. Herein, lentiviral delivery of CBFβ‐specific shRNAs was used to achieve a 95% reduction of CBFβ in an ovarian cancer cell line. This drastic reduction in CBFβ expression resulted in growth inhibition that was not associated with a cell cycle block or an increase in apoptosis. However, CBFβ silencing resulted in increased autophagy and production of reactive oxygen species (ROS). Since sphingolipid and ceramide metabolism regulates non‐apoptotic cell death, autophagy, and ROS production, fumonsin B1 (FB1), an inhibitor of ceramide synthase, was used to alter ceramide production in the CBFβ‐silenced cells. FB1 treatment inhibited the CBFβ‐dependent increase in autophagy and provided a modest increase in cell survival. To document alterations to sphingolipids in the CBFβ‐silenced cells, ceramide, and lactosylceramide levels were directly examined by mass spectrometry. Substantial increases in ceramide species and decreases in lactosylceramides were identified. Altogether, this report provides evidence that CBF transcriptional pathways control cellular survival, at least in part, through sphingolipid metabolism. J. Cell. Physiol. 228: 2350–2364, 2013. © 2013 Wiley Periodicals, Inc.

  August 23, 2013   doi: 10.1002/jcp.24406   open full text
• Nitric oxide regulation of Na,K‐ATPase activity in ocular ciliary epithelium involves Src family kinase.
  Mohammad Shahidullah, Amritlal Mandal, Guojun Wei, Nicholas A Delamere.
  Journal of Cellular Physiology. August 22, 2013
  The nitric oxide (NO) donor sodium nitroprusside (SNP) is known to reduce aqueous humor (AH) secretion in the isolated porcine eye. Previously, SNP was found to inhibit Na,K‐ATPase activity in nonpigmented ciliary epithelium (NPE), AH‐secreting cells, through a cGMP/protein kinase G (PKG)‐mediated pathway. Here we show Src family kinase (SFK) activation in the Na,K‐ATPase activity response to SNP. Ouabain‐sensitive 86Rb uptake was reduced by >35% in cultured NPE cells exposed to SNP (100 µM) or exogenously added cGMP (8‐Br‐cGMP) (100 µM) and the SFK inhibitor PP2 (10 µM) prevented the response. Ouabain‐sensitive ATP hydrolysis was reduced by ∼40% in samples detected in material obtained from SNP‐ and 8‐Br‐cGMP‐treated cells following homogenization, pointing to an intrinsic change of Na,K‐ATPase activity. Tyrosine‐10 phosphorylation of Na,K‐ATPase α1 subunit was detected in SNP and L‐arginine‐treated cells and the response prevented by PP2. SNP elicited an increase in cell cGMP. Cells exposed to 8‐Br‐cGMP displayed SFK activation (phosphorylation) and inhibition of both ouabain‐sensitive 86Rb uptake and Na,K‐ATPase activity that was prevented by PP2. SFK activation, which also occurred in SNP‐treated cells, was suppressed by inhibitors of soluble guanylate cyclase (ODQ;10 µM) and protein kinase G (KT5823;1 µM). SNP and 8‐Br‐cGMP also increased phosphorylation of ERK1/2 and p38 MAPK and the response prevented by PP2. However, U0126 did not prevent SNP or 8‐Br‐cGMP‐induced inhibition of Na,K‐ATPase activity. Taken together, the results suggest that NO activates guanylate cyclase to cause a rise in cGMP and subsequent PKG‐dependent SFK activation. Inhibition of Na,K‐ATPase activity depends on SFK activation. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 22, 2013   doi: 10.1002/jcp.24454   open full text
• Advances in androgen receptor targeted therapy for prostate cancer.
  Alia Ahmed, Shadan Ali, Fazlul H. Sarkar.
  Journal of Cellular Physiology. August 22, 2013
  Prostate Cancer (PCa) is the second leading cause of cancer death in men. Current research findings suggest that the androgen receptor (AR) and its signaling pathway contribute significantly to the progression of metastatic PCa. The AR is a ligand activated transcription factor, where androgens such as testosterone (T) and dihydroxytestosterone (DHT) act as the activating ligands. However in many metastatic PCa, the AR functions promiscuously and is constitutively active through multiple mechanisms. Inhibition of enzymes that take part in androgen synthesis or synthesizing antiandrogens that can inhibit the AR are two popular methods of impeding the androgen receptor signaling axis; however, the inhibition of androgen‐independent activated AR function has not yet been fully exploited. This article focuses on the development of emerging novel agents that act at different steps along the androgen‐AR signaling pathway to help improve the poor prognosis of PCa patients. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 22, 2013   doi: 10.1002/jcp.24456   open full text
• Dynamic measurement of ATP in single oocytes: Impact of stage of maturation and cumulus cells on ATP levels and rates of consumption.
  Caroline M Dalton, Gyorgy Szabadkai, John Carroll.
  Journal of Cellular Physiology. August 22, 2013
  Mitochondria provide the primary source of ATP in the oocyte and early embryo and mitochondrial dysfunction and deficit of mitochondria‐derived ATP has been linked to suboptimal developmental competence. We have undertaken a study of ATP in the maturing mouse oocyte using a novel recombinant FRET based probe, AT1.03. We show that AT1.03 can be successfully used to monitor cytosolic ATP levels in single live oocytes over extended time periods. We find that ATP levels undergo dynamic changes associated with specific maturational events and that oocytes display altered rates of ATP consumption at different stages of maturation. Cumulus enclosed oocytes have a higher ATP level during maturation than denuded oocytes and this can be abolished by inhibition of gap junctional communication between the oocyte and cumulus cells. Our work uses a new approach to shed light on regulation of ATP levels and ATP consumption during oocyte maturation. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 22, 2013   doi: 10.1002/jcp.24457   open full text
• A β1/2 adrenergic receptor‐sensitive intracellular signaling pathway modulates CCL2 production in cultured spinal astrocytes.
  Norimitsu Morioka, Hiromi Abe, Ryosuke Araki, Naoki Matsumoto, Fan Fan Zhang, Yoki Nakamura, Kazue Hisaoka‐Nakashima, Yoshihiro Nakata.
  Journal of Cellular Physiology. August 22, 2013
  The phosphorylation of c‐jun N‐terminal kinase (JNK) and the subsequent production of C‐C chemokine CCL2 (monocyte chemoattractant protein; MCP‐1) in spinal astrocytes contribute to the initiation of neurological disorders including chronic pain. Astrocytes express neurotransmitter receptors which could be targeted to ameliorate neurological disorders. In the current study, the involvement of the β‐adrenergic system in the regulation of JNK activity and CCL2 production after stimulation with tumor necrosis factor (TNF)‐α, one of many initiators of neuroinflammation, was elucidated. Treatment of cultured spinal astrocytes with isoproterenol (a β‐adrenergic receptor agonist; 1 µM) reduced both TNF‐α‐induced JNK1 phosphorylation, as observed by Western blotting, and the subsequent increase of both CCL2 mRNA expression and CCL2 production, which were measured by real time‐PCR and ELISA, respectively. The effects of isoproterenol were completely blocked by pretreatment with either propranolol (a β‐adrenoceptor antagonist) or H89 (a protein kinase A (PKA) inhibitor). The current study revealed that the regulation of glycogen synthase kinase‐3β (GSK‐3β) activity is a crucial factor in the inhibitory action of isoproterenol. The TNF‐α‐induced JNK1 phosphorylation was significantly blocked by treatment with GSK‐3β inhibitors (either LiCl or TWS119), and stimulation of β‐adrenergic receptors induced the inhibition of GSK‐3β through the phosphorylation of Ser9. Moreover, treatment with isoproterenol markedly suppressed the TNF‐α‐induced increase of CCL2 mRNA expression and CCL2 production through a β‐adrenergic receptor‐PKA pathway mediated by GSK‐3β regulation. Thus, activation of β1/2 adrenergic receptors expressed in spinal astrocytes could be a novel method of moderating neurological disorders with endogenous catecholamines or selective agonists. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 22, 2013   doi: 10.1002/jcp.24452   open full text
• P2X7 receptors in neurohypophysial terminals: Evidence for their role in arginine‐vasopressin secretion.
  Adolfo E. Cuadra, Edward E. Custer, Elizabeth L. Bosworth, José R. Lemos.
  Journal of Cellular Physiology. August 22, 2013
  Arginine‐vasopressin (AVP) plays a major role in maintaining cardiovascular function and related pathologies. The mechanism involved in its release into the circulation is complex and highly regulated. Recent work has implicated the purinergic receptor, P2X7R, in a role for catecholamine‐enhanced AVP release in the rat hypothalamic‐neurohypophysial (NH) system. However, the site of P2X7R action in this endocrine system, and whether or not it directly mediates release in secretory neurons have not been determined. We hypothesized that the P2X7R is expressed and mediates AVP release in NH terminals. P2X7R function was first examined by patch‐clamp recordings in isolated NH terminals. Results revealed that subpopulations of isolated terminals displayed either high ATP‐sensitivity or low ATP‐sensitivity, the latter of which was characteristic of the rat P2X7R. Additional recordings showed that terminals showing sensitivity to the P2X7R‐selective agonist, BzATP, were further inhibited by P2X7R selective antagonists, AZ10606120 and brilliant blue‐G. In confocal micrographs from tissue sections and isolated terminals of the NH P2X7R‐immunoreactivity was found to be localized in plasma membranes. Lastly, the role of P2X7R on AVP release was tested. Our results showed that BzATP evoked sustained AVP release in NH terminals, which was inhibited by AZ10606120. Taken together, our data lead us to conclude that the P2X7R is expressed in NH terminals and corroborates its role in AVP secretion. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 22, 2013   doi: 10.1002/jcp.24453   open full text
• EBV‐encoded LMP‐1 sensitizes nasopharyngeal carcinoma cells to genotoxic drugs by down‐regulating Cabin1 expression.
  Pu‐Yuan Chang, Zchong‐Zcho Wu, Nian‐Kang Sun, Chuck C.‐K. Chao.
  Journal of Cellular Physiology. August 13, 2013
  The oncogenic latent membrane protein 1 (LMP1) of Epstein–Barr virus (EBV) is involved in the pathogenesis of human nasopharyngeal carcinoma (NPC) and lymphoma. We and other authors have shown earlier that LMP1 induces apoptosis and inhibits xenograft tumor growth in mice, but the mechanism underlying these processes has not been investigated so far. In the present study, we show that knockdown of LMP1 renders the EBV‐positive NPC cell line CG‐1 resistant to various genotoxic drugs (cisplatin, etoposide, and adriamycin). LMP1 inhibits the expression of Cabin1, a Ca2+ regulated protein shown earlier to inhibit calcineurin. Knockdown of Cabin1 with small hairpin RNA sensitizes CG‐1 cells to genotoxic drugs. In contrast, LMP1 overexpression reduces Cabin1 level and renders both CG‐1 cells and EBV‐negative NPC cell lines sensitive to cisplatin. The JNK and ERK pathways are required for LMP1‐induced suppression of Cabin1 at the transcriptional level. Chromatin immunoprecipitation assays further confirm that the JNK‐activated transcription factor AP‐1 mediates the LMP1‐induced down‐regulation of Cabin1 gene expression. LMP1 knockdown also increases the resistance of xenograph tumors to cisplatin in mice, therefore confirming the relevance of our findings in vivo. This study reveals the molecular mechanism underlying the pro‐apoptotic activity of LMP1 during cisplatin‐based NPC chemotherapy. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 13, 2013   doi: 10.1002/jcp.24448   open full text
• Interleukin‐17A increases cell migration and MMP‐1 expression in human periodontal ligament fibroblasts via p38 MAPK/NF‐κB ‐dependent pathway.
  Yan Wu, Lingxin Zhu, Lingshuang Liu, Jie Zhang, Bin Peng.
  Journal of Cellular Physiology. August 08, 2013
  Interleukin‐17 (IL‐17) is a cytokine secreted predominantly by Th17 cells. Although IL‐17 is primarily associated with the induction of tissue inflammation, the other biological functions of IL‐17, including its wound‐healing functions, have yet to be thoroughly explored. Fibroblast proliferation and migration play essential roles in periodontal wound‐healing responses. In this study, we report that IL‐17A can increase the migration and expression of matrix metalloproteinase (MMP)‐1 in human periodontal ligament (PDL) fibroblasts but has no effect on PDL fibroblast proliferation. IL‐17A‐induced MMP‐1 expression led to cell migration, which was attenuated by pre‐treatment with IL‐17 receptor neutralizing antibody and small interfering RNA (siRNA) for MMP‐1. The IL‐17A‐induced cell migration was also attenuated by its tissue inhibitor of matrix metalloproteinase (TIMP)‐1. In addition, a p38 mitogen‐activated protein kinase (MAPK) inhibitor (SB203580) inhibited IL‐17A‐induced increase of the migration and MMP‐1 upregulation of PDL fibroblasts. The involvement of p38 MAPK in IL‐17A‐induced MMP‐1 expression and cell migration was further confirmed by transfection of p38α siRNA. A nuclear factor kappaB ((NF‐κB) inhibitor (pyrrolidine dithiocarbamate) also suppressed the cell migration and MMP‐1 expression enhanced by IL‐17A. Moreover, transfection with p38α siRNA inhibited IL‐17A‐induced NF‐κB nuclear translocation as well as NF‐κB binding activity. Our results suggest that IL‐17A enhances the migration of PDL fibroblasts by increasing MMP‐1 expression through the IL‐17 receptor, p38 MAPK, and NF‐κB signal transduction pathways. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 08, 2013   doi: 10.1002/jcp.24444   open full text
• Extreme thermal noxious stimuli induce pain responses in zebrafish larvae.
  Valentina Malafoglia, Marco Colasanti, William Raffaeli, Darius Balciunas, Antonio Giordano, Gianfranco Bellipanni.
  Journal of Cellular Physiology. August 08, 2013
  Exposing tissues to extreme high or low temperature leads to burns. Burned animals sustain several types of damage, from the disruption of the tissue to degeneration of axons projecting through muscle and skin. Such damage causes pain due to both inflammation and axonal degeneration (neuropathic‐like pain). Thus, the approach to cure and alleviate the symptoms of burns must be twofold: rebuilding the tissue that has been destroyed and alleviating the pain derived from the burns. While tissue regeneration techniques have been developed, less is known on the treatment of the induced pain. Thus, appropriate animal models are necessary for the development of the best treatment for pain induced in burned tissues. We have developed a methodology in the zebrafish aimed to produce a new animal model for the study of pain induced by burns. Here we show that two events linked to the onset of burn‐induced inflammation and neuropathic‐like pain in mammals, degeneration of axons innervating the affected tissues and over‐expression of specific genes in sensory tissues, are conserved from zebrafish to mammals. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 08, 2013   doi: 10.1002/jcp.24447   open full text
• Emerging roles of microRNA in modulating cell‐death processes in malignant glioma.
  Silvia Palumbo, Clelia Miracco, Luigi Pirtoli, Sergio Comincini.
  Journal of Cellular Physiology. August 08, 2013
  MicroRNAs (miRNAs) are small noncoding RNA molecules that regulate protein expression by cleaving or repressing the translation of target mRNAs. In mammals, their function mainly represses the mRNA transcripts via imperfect complementary sequences in the 3'UTR of target mRNAs. Several miRNAs have been recently reported to be involved in modulation of different genes in tumors, including glioblastoma, the most frequent brain tumor in adults. Despite the improvements in treatments, survival of patients remains poor, and glioblastoma is one of the most lethal form of human cancer. To define novel strategies against this tumor, emerging research investigated miRNAs involvement in glioblastoma. In particular, this review is focused on miRNAs involved on the two principal programmed cell‐death, apoptosis and autophagy, recently described from the literature. Moreover, the discovery of miRNAs role in glioma cell‐death pathways has also revealed a new category of therapeutic targets, fundamental for this kind of tumor. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  August 08, 2013   doi: 10.1002/jcp.24446   open full text
• Blood Type B antigen modulates cell migration through regulating cdc42 expression and activity in HaCaT cells.
  Ji‐Yong Jung, Jang‐Hee Oh, Dong Hun Lee, Serah Lee, Jin Ho Chung.
  Journal of Cellular Physiology. July 25, 2013
  ABO blood group is determined by carbohydrate antigens, called ABH antigens. It has been known that the change of carbohydrate antigen expression, including ABH antigens, has correlation with the tumor metastasis and survival; however, the exact mechanism remains to be elucidated. ABH antigens are expressed not only in blood cells but also in several tissues. In epidermis, ABH antigen is expressed in the uppermost spinous and granular layer. We investigated the role of ABH antigens on the cell migration of HaCaT keratinocytes, which express B antigen. Knock‐down of B antigen expression by small interference RNA of FUT1 inhibited HaCaT cell migration. At that time, we found that lamellipodia and actin fiber were also reduced by knock‐down of B antigen expression. The transcription of cdc42, a kind of Rho GTPase which plays a key role in actin polymerization, was reduced by down‐regulated B antigen expression. Furthermore, the reduced B antigen expression also inhibited the interaction of cdc42 and N‐WASP. Collectively, our data provide a clue how ABH antigens regulate the cell migration mechanism. J. Cell. Physiol. 228: 2243–2251, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24393   open full text
• Functional role of Runx3 in the regulation of aggrecan expression during cartilage development.
  Nathan A. Wigner, Do Y. Soung, Thomas A. Einhorn, Hicham Drissi, Louis C. Gerstenfeld.
  Journal of Cellular Physiology. July 25, 2013
  Runx2 and Runx3 are known to be expressed in the growth plate during endochondral bone formation. Here we addressed the functional role of Runx3 as distinct from Runx2 by using two models of postnatal bone repair: fracture healing that proceeds by an endochondral process and marrow ablation that proceeds by only an intramembranous process. Both Runx2 and Runx3 mRNAs were differentially up regulated during fracture healing. In contrast, only Runx2 showed increased expression after marrow ablation. During fracture healing, Runx3 was expressed earlier than Runx2, was concurrent with the period of chondrogenesis, and coincident with maximal aggrecan expression a protein associated with proliferating and permanent cartilage. Immunohistological analysis showed Runx3 protein was also expressed by chondrocytes in vivo. In contrast, Runx2 was expressed later during chondrocyte hypertrophy, and primary bone formation. The functional activities of Runx3 during chondrocyte differentiation were assessed by examining its regulatory actions on aggrecan gene expression. Aggrecan mRNA levels and aggrecan promoter activity were enhanced in response to the over‐expression of either Runx2 and Runx3 in ATDC5 chondrogenic cell line, while sh‐RNA knocked down of each Runx protein showed that only Runx3 knock down specifically suppressed aggrecan mRNA expression and promoter activity. ChIP assay demonstrated that Runx3 interactions were selective to sites within the aggrecan promoter and were only observed during early periods of chondrogenesis before hypertrophy. Our studies suggest that Runx3 positively regulates aggrecan expression and suggest that its function is more limited to cartilage development than to bone. In aggregate these data further suggest that the various members of the Runx transcription factors are involved in the coordination of chondrocyte development, maturation, and hypertrophy during endochondral bone formation. J. Cell. Physiol. 228: 2232–2242, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24396   open full text
• FUS/TLS assembles into stress granules and is a prosurvival factor during hyperosmolar stress.
  Reddy Ranjith K. Sama, Catherine L. Ward, Laura J. Kaushansky, Nathan Lemay, Shinsuke Ishigaki, Fumihiko Urano, Daryl A. Bosco.
  Journal of Cellular Physiology. July 25, 2013
  FUsed in Sarcoma/Translocated in LipoSarcoma (FUS/TLS or FUS) has been linked to several biological processes involving DNA and RNA processing, and has been associated with multiple diseases, including myxoid liposarcoma and amyotrophic lateral sclerosis (ALS). ALS‐associated mutations cause FUS to associate with stalled translational complexes called stress granules under conditions of stress. However, little is known regarding the normal role of endogenous (non‐disease linked) FUS in cellular stress response. Here, we demonstrate that endogenous FUS exerts a robust response to hyperosmolar stress induced by sorbitol. Hyperosmolar stress causes an immediate re‐distribution of nuclear FUS to the cytoplasm, where it incorporates into stress granules. The redistribution of FUS to the cytoplasm is modulated by methyltransferase activity, whereas the inhibition of methyltransferase activity does not affect the incorporation of FUS into stress granules. The response to hyperosmolar stress is specific, since endogenous FUS does not redistribute to the cytoplasm in response to sodium arsenite, hydrogen peroxide, thapsigargin, or heat shock, all of which induce stress granule assembly. Intriguingly, cells with reduced expression of FUS exhibit a loss of cell viability in response to sorbitol, indicating a prosurvival role for endogenous FUS in the cellular response to hyperosmolar stress. J. Cell. Physiol. 228: 2222–2231, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24395   open full text
• Altered bone development and turnover in transgenic mice over‐expressing Lipocalin‐2 in bone.
  Delfina Costa, Edoardo Lazzarini, Barbara Canciani, Alessandra Giuliani, Raffaele SpanÒ, Katia Marozzi, Adrian Manescu, Ranieri Cancedda, Sara Tavella.
  Journal of Cellular Physiology. July 25, 2013
  Lipocalin‐2 (LCN2) is a protein largely expressed in many tissues, associated with different biological phenomena such as cellular differentiation, inflammation and cancer acting as a survival/apoptotic signal. We found that LCN2 was expressed during osteoblast differentiation and we generated transgenic (Tg) mice over‐expressing LCN2 in bone. Tg mice were smaller and presented bone microarchitectural changes in both endochondral and intramembranous bones. In particular, Tg bones displayed a thinner layer of cortical bone and a decreased trabecular number. Osteoblast bone matrix deposition was reduced and osteoblast differentiation was slowed‐down. Differences were also observed in the growth plate of young transgenic mice where chondrocyte displayed a more immature phenotype and a lower proliferation rate. In bone marrow cell cultures from transgenic mice, the number of osteoclast progenitors was increased whereas in vivo it was increased the number of mature osteoclasts expressing tartrate‐resistant acid phosphatase (TRAP). Finally, while osteoprotegerin (OPG) levels remained unchanged, the expression of the conventional receptor activator of nuclear factor‐κB ligand (RANKL) and of the IL‐6 was enhanced in Tg mice. In conclusion, we found that LCN2 plays a role in bone development and turnover having both a negative effect on bone formation, by affecting growth plate development and interfering with osteoblast differentiation, and a positive effect on bone resorption by enhancing osteoclast compartment. J. Cell. Physiol. 228: 2210–2221, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24391   open full text
• Therapeutic potential of a pyridoxal‐based vanadium(IV) complex showing selective cytotoxicity for cancer versus healthy cells.
  Maria Strianese, Anna Basile, Antonio Mazzone, Silvana Morello, Maria Caterina Turco, Claudio Pellecchia.
  Journal of Cellular Physiology. July 25, 2013
  Vanadium compounds can exert anticancer effects, partly due to inhibition of tyrosine phosphatases. Here, we report the effect of N,N′‐ethylenebis (pyridoxylideneiminato) vanadium (IV) complex (Pyr2enV(IV)), that induced 93% and 57% of cell mortality in A375 (human melanoma) and A549 (human lung carcinoma) cells, respectively; the mortality was <24% in other cancer cell lines and in human normal epidermal keratinocytes, lung cells and peripheral blood mononuclear cells. The mechanism of Pyr2enV(IV) effect relied on apoptosis induction; this was triggered by ROS increase, followed by mitochondrial membrane depolarization. Indeed, the addition of N‐acetyl cysteine to cell cultures abated Pyr2enV(IV)‐induced apoptosis. These results disclose the pro‐apoptotic activity of Pyr2enV(IV) and its mechanism, relying on intracellular ROS increase. J. Cell. Physiol. 228: 2202–2209, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24385   open full text
• Regulation of the delta and alpha epithelial sodium channel (ENaC) by ubiquitination and Nedd8.
  Kevin Ly, C. Joy McIntosh, Wolfgang Biasio, Yongfeng Liu, Ying Ke, Diane R. Olson, John H. Miller, Rachel Page, Peter M. Snyder, Fiona J. McDonald.
  Journal of Cellular Physiology. July 25, 2013
  The δ epithelial sodium channel (δENaC) is a proton‐activated, sodium‐selective, amiloride‐sensitive ion channel in the ENaC/degenerin family of ion channels involved in blood pressure regulation and mechanosensation. Other ENaC family members are subject to ubiquitin modification leading to internalization from the cell surface, and degradation of the channel. Here, we show that δENaC is also modified by ubiquitin on three intracellular lysine residues. Absence of these lysines abolished ubiquitin modification of δENaC and increased cell surface levels of δENaC. Although the HECT‐domain ubiquitin ligase Nedd4‐2 reduced amiloride‐sensitive current generated by δβγENaC‐containing channels, δENaC does not contain a binding site for Nedd4‐2; therefore, this effect is probably mediated by the βγENaC subunits. Nedd8, a ubiquitin‐like protein that regulates RING‐domain E3 ubiquitin ligases, promoted δENaC ubiquitination, decreased both the intracellular and cell surface δENaC populations, and decreased δβγENaC amiloride‐sensitive short circuit current (Isc‐amiloride) in a mammalian epithelium. Nedd8 also promoted α− and γENaC ubiquitination, decreased the cell surface pools, and decreased αβγENaC Isc‐amiloride. Conversely, XIAP, a single subunit RING E3 ligase, decreased ubiquitinated δENaC, increased the δENaC cell surface pool and increased δβγENaC Isc‐amiloride. Therefore δ− and α − βγENaC channel function may be influenced by RING‐domain E3 ubiquitin ligases. J. Cell. Physiol. 228: 2190–2201, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24390   open full text
• PHLP2 is essential and plays a role in ciliogenesis and microtubule assembly in Tetrahymena thermophila.
  Cezary Bregier, Lucja KrzemieŃ‐Ojak, Dorota WŁOga, Maria Jerka‐Dziadosz, Ewa Joachimiak, Katarzyna Batko, Iwona Filipiuk, Urszula ŚMietanka, Jacek Gaertig, StanisŁAw Fabczak, Hanna Fabczak.
  Journal of Cellular Physiology. July 25, 2013
  Recent studies have implicated the phosducin‐like protein‐2 (PHLP2) in regulation of CCT, a chaperonin whose activity is essential for folding of tubulin and actin. However, the exact molecular function of PHLP2 is unclear. Here we investigate the significance of PHLP2 in a ciliated unicellular model, Tetrahymena thermophila, by deleting its single homolog, Phlp2p. Cells lacking Phlp2p became larger and died within 96 h. Overexpressed Phlp2p‐HA localized to cilia, basal bodies, and cytosol without an obvious change in the phenotype. Despite similar localization, overexpressed GFP‐Phlp2p caused a dominant‐negative effect. Cells overproducing GFP‐Phlp2p had decreased rates of proliferation, motility and phagocytosis, as compared to wild type cells or cells overproducing a non‐tagged Phlp2p. Growing GFP‐Phlp2p‐overexpressing cells had fewer cilia and, when deciliated, failed to regenerate cilia, indicating defects in cilia assembly. Paclitaxel‐treated GFP‐Phlp2p cells failed to elongate cilia, indicating a change in the microtubules dynamics. The pattern of ciliary and cytosolic tubulin isoforms on 2D gels differed between wild type and GFP‐Phlp2p‐overexpressing cells. Thus, in Tetrahymena, PhLP2 is essential and under specific experimental conditions its activity affects tubulin and microtubule‐dependent functions including cilia assembly. J. Cell. Physiol. 228: 2175–2189, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24384   open full text
• 1α,25‐dihydroxyvitamin D3 stimulates activin A production to fine‐tune osteoblast‐induced mineralization.
  V.J. Woeckel, B.C.J. van der Eerden, M. Schreuders‐Koedam, M. Eijken, J.P.T.M. Van Leeuwen.
  Journal of Cellular Physiology. July 25, 2013
  In healthy bones, mineralization has to be tightly controlled to avoid pathological phenotypes. In this study, we investigated interactions between 1α,25(OH)2D3 (1,25D3) and activin A in the regulation of osteoblast induced mineralization. In human osteoblast cultures, we demonstrated that besides stimulation of mineralization, 1,25D3 also induced activin A, a strong inhibitor of mineralization. Simultaneously, follistatin (FST), the natural antagonist of activin A, was down‐regulated by1,25D3. This resulted in an increase in activin A activity during 1,25D3 treatment. We also showed that in 1,25D3‐treated osteoblasts, mineralization can be further increased when activin A activity was abrogated by adding exogenous FST. This observation implies that, besides stimulation of mineralization, 1,25D3 also controls activin A‐mediated inhibition of mineralization. Besides activin A, 1,25D3 also induces osteocalcin (BGLAP), another inhibitor of mineralization. Warfarin, which has been shown to inactivate osteocalcin, increased 1,25D3‐induced mineralization. Interaction between these two systems became evident from the synergistic increase in BGLAP expression upon blocking activin activity in 1,25D3‐treated cultures. In conclusion, we demonstrate that 1,25D3 stimulation of mineralization by human osteoblasts is suppressed by concomitant induction of inhibitors of mineralization. Mineralization induction by 1,25D3 may actually be controlled via interplay with activin A and osteocalcin. Finally, this complex regulation of mineralization substantiates the significance of tight control of mineralization to prevent excessive mineralization and consequently reduction in bone quality and strength. J. Cell. Physiol. 228: 2167–2174, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24388   open full text
• Culture under low physiological oxygen conditions improves the stemness and quality of induced pluripotent stem cells.
  Chao‐Wan Guo, Miho Kawakatsu, Marie Idemitsu, Yoshishige Urata, Shinji Goto, Yusuke Ono, Kimikazu Hamano, Tao‐Sheng Li.
  Journal of Cellular Physiology. July 25, 2013
  The ex vivo expansion of stem cells under low physiological oxygen (O2) conditions has been demonstrated to improve the stemness and genomic stability of the cells. We investigated whether low‐oxygen culture would be beneficial for the culture of induced pluripotent stem (iPS) cells. Two human iPS cell lines (201B7 and 253G1) were used for the experiments. Cells expanded from a single colony of each cell line were initiated for culture in 2.5% O2, 5% O2, or 20% O2 and maintained for 2 months in parallel. The levels of intracellular and mitochondrial reactive oxygen species did not differ between the cells cultured under different conditions. More colonies of uniformly smaller size were observed at 2.5% and 5% O2 than at 20% O2. All of these iPS colonies that expanded under the various oxygen conditions stained positively for Oct3/4, Nanog, SSEA‐4, and ALP. However, Western blot analysis showed that the iPS cells cultured at 2.5% and 5% O2 expressed significantly more Nanog but less 53BP1 than those cultured at 20% O2. Data from an array CGH showed no significant chromosomal abnormalities, although some genes involved in cellular and metabolic processes were amplified in the low oxygen culture, particularly at 2.5% O2. Our data suggest that low physiological oxygen culture could improve the stemness and quality of iPS cells, a result that might be associated with the amplification of genes involved in metabolic and cellular processes. Long‐term culture will be necessary to confirm whether low physiological oxygen levels also improve genomic stability. J. Cell. Physiol. 228: 2159–2166, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24389   open full text
• FGF‐2 enhances Runx‐2/Smads nuclear localization in BMP‐2 canonical signaling in osteoblasts.
  Dimitrios Agas, Maria Giovanna Sabbieti, Luigi Marchetti, Liping Xiao, Marja M. Hurley.
  Journal of Cellular Physiology. July 25, 2013
  Bone morphogenetic protein 2 (BMP‐2) is one of the most potent regulators of osteoblast differentiation and bone formation. R‐Smads (Smads 1/5/8) are the major transducers for BMPs receptors and, once activated, they are translocated in the nucleus regulating transcription target genes by interacting with various transcription factors. Runx‐2 proteins have been shown to interact through their C‐terminal segment with Smads and this interaction is required for in vivo osteogenesis. In particular, recruitment of Smads to intranuclear sites is Runx‐2 dependent, and Runx‐2 factor may accommodate the dynamic targeting of signal transducer to active transcription sites. Previously, we have shown, by in vitro and in vivo experiments, that BMP‐2 up‐regulated FGF‐2 which is important for the maximal responses of BMP‐2 in bone. In this study, we found that endogenous FGF2 is necessary for BMP‐2 induced nuclear accumulation and co‐localization of Runx‐2 and phospho‐Smads1/5/8, while Runx/Smads nuclear accumulation and co‐localization was reduced in Fgf2−/− osteoblasts. Based on these novel data, we conclude that the impaired nuclear accumulation of Runx‐2 in Fgf2−/− osteoblasts reduces R‐Smads sub‐nuclear targeting with a consequent decreased expression of differentiating markers and impaired bone formation in Fgf2 null mice. J. Cell. Physiol. 228: 2149–2158, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24382   open full text
• Suppression of interactions between prostate tumor cell‐surface integrin and endothelial ICAM‐1 by simvastatin inhibits micrometastasis.
  Belal AL‐Husein, Anna Goc, Payaningal R. Somanath.
  Journal of Cellular Physiology. July 25, 2013
  Cancer micrometastasis relies on the ability of cancer cells to secrete angiogenic modulators, to interact with the vascular endothelium, and to overcome the resistance offered by the endothelial‐barrier. Being an essential step prior to metastasis, blockage of micrometastasis can have potential applications in cancer therapy and metastasis prevention. Due to poorly known molecular mechanisms leading to micrometastasis, developing therapeutic strategies to target prostate cancer utilizing drugs that block micrometastasis is far from reality. Here, we demonstrate the potential benefits of simvastatin in the inhibition of prostate cancer micrometastasis and reveal the novel molecular mechanisms underlying this process. First, we showed that simvastatin inhibited the ability of human PC3 prostate cancer cells for transendothelial migration in vitro. Second, our data indicated that simvastatin modulates the expression of tumor‐derived factors such as angiopoietins and VEGF‐A at the mRNA and protein levels by the PC3 cells, thus preventing endothelial‐barrier disruption. Third, simvastatin directly activated endothelial cells and enhances endothelial‐barrier resistance. Apart from this, our study revealed that simvastatin‐mediated effect on PC3 micrometastasis was mediated through inhibition of integrin αvβ3 activity and suppression of interaction between prostate cancer cell integrin αvβ3 with endothelial ICAM‐1. J. Cell. Physiol. 228: 2139–2148, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24381   open full text
• Polycyclic aromatic hydrocarbons—induced ROS accumulation enhances mutagenic potential of T‐antigen from human polyomavirus JC.
  Anna Wilk, Piotr Waligórski, Adam Lassak, Himanshu Vashistha, David Lirette, David Tate, Arnold H. Zea, Shahriar Koochekpour, Paulo Rodriguez, Leonard G. Meggs, John J. Estrada, Augusto Ochoa, Krzysztof Reiss.
  Journal of Cellular Physiology. July 25, 2013
  Polycyclic aromatic hydrocarbons (PAHs) are the products of incomplete combustion of organic materials, which are present in cigarette smoke, deep‐fried food, and in natural crude oil. Since PAH‐metabolites form DNA adducts and cause oxidative DNA damage, we asked if these environmental carcinogens could affect transforming potential of the human Polyomavirus JC oncoprotein, T‐antigen (JCV T‐antigen). We extracted DMSO soluble PAHs from Deepwater Horizon oil spill in the Gulf of Mexico (oil‐PAHs), and detected several carcinogenic PAHs. The oil‐PAHs were tested in exponentially growing cultures of normal mouse fibroblasts (R508), and in R508 stably expressing JCV T‐antigen (R508/T). The oil‐PAHs were cytotoxic only at relatively high doses (1:50–1:100 dilution), and at 1:500 dilution the growth and cell survival rates were practically unaffected. This non‐toxic dose triggered however, a significant accumulation of reactive oxygen species (ROS), caused oxidative DNA damage and the formation of DNA double strand breaks (DSBs). Although oil‐PAHs induced similar levels of DNA damage in R508 and R508/T cells, only T‐antigen expressing cells demonstrated inhibition of high fidelity DNA repair by homologous recombination (HRR). In contrast, low‐fidelity repair by non‐homologous end joining (NHEJ) was unaffected. This potential mutagenic shift between DNA repair mechanisms was accompanied by a significant increase in clonal growth of R508/T cells chronically exposed to low doses of the oil‐PAHs. Our results indicate for the first time carcinogenic synergy in which oil‐PAHs trigger oxidative DNA damage and JCV T‐antigen compromises DNA repair fidelity. J. Cell. Physiol. 228: 2127–2138, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24375   open full text
• Small GTPase Rap1 regulates cell migration through regulation of small GTPase RhoA activity in response to transforming growth factor‐β1.
  Mi‐Young Moon, Hee‐Jun Kim, Jae‐Gyu Kim, Jae‐Yong Lee, Jaebong Kim, Sung‐Chan Kim, Ihn‐Geun Choi, Pyeung‐Hyeun Kim, Jae‐Bong Park.
  Journal of Cellular Physiology. July 25, 2013
  Transforming growth factor (TGF)‐β1 regulates diverse cellular functions. Particularly, TGF‐β1 induces monocyte migration to sites of injury or inflammation in early period, whereas TGF‐β1 inhibits cell migration in late phase. In this study, we attempted to understand how TGF‐β1 suppresses cell migration in late phase. We found that TGF‐β1 of short exposure induces the production of chemokines, such as macrophage inflammatory protein (MIP)‐1α, by Raw 264.7 cells. However, knock‐down of small GTPase RhoA by sh‐RhoA inhibited the production of MIP‐1α and macrophage migration, suggesting that RhoA is essential for expression of this chemokine. An activator of Epac (exchange proteins directly activated by cAMP; a guanine nucleotide exchange factor of Rap1), 8CPT‐2Me‐cAMP which leads to Rap1 activation abrogated MIP‐1α expression and macrophage migration. Indeed, GTP‐RhoA and GTP‐Rap1 levels were reciprocally regulated in a time‐dependent manner following TGF‐β1 stimulation. 8CPT‐2Me‐cAMP suppressed GTP‐RhoA levels, whereas si‐Rap1 augmented GTP‐RhoA levels and cell migration. TGF‐β1 produced cAMP in late period and si‐RNAs of Epac1 and Epac2 reduced GTP‐Rap1 levels leading to promotion of GTP‐RhoA levels. Furthermore, si‐RNA of ARAP3 (Rap‐dependent RhoGAP) increased GTP‐RhoA level and cell migration. Therefore, we propose the mechanism that prolonged TGF‐β1 treatment produce cAMP, which activates sequentially Epac, Rap1 and ARAP3, resulting in suppression of RhoA, chemokine expression, and macrophage migration. Contrary to the general concept that Rap1 stimulates cell migration, we demonstrated in this study that Rap1 inhibits cell migration by suppression of RhoA activity in response to TGF‐β1. J. Cell. Physiol. 228: 2119–2126, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24383   open full text
• Differentiation of human adipose stem cells into neural phenotype by neuroblastoma‐ or olfactory ensheathing cells‐conditioned medium.
  Debora Lo Furno, Rosalia Pellitteri, Adriana C.E. Graziano, Rosario Giuffrida, Carlo Vancheri, Elisa Gili, Venera Cardile.
  Journal of Cellular Physiology. July 25, 2013
  Olfactory ensheathing cells (OECs) are known to be capable of continuous neurogenesis throughout lifetime and are a source of multiple trophic factors important in central nervous system regeneration. B104 neuroblastoma cells are recognized to induce differentiation of neural stem cells into oligodendrocyte precursor cells. Therefore, the aim of this study was to verify if conditioned medium (CM) obtained from OECs or B104 cells was capable of inducing differentiation of adipose tissue‐derived mesenchymal stem cells (AT‐MSCs) to a neuronal phenotype. In order to this goal, immunocytochemical procedures and flow cytometry analysis were used and some neural markers, as nestin, protein gene product 9.5 (PGP 9.5), microtubule‐associated protein 2 (MAP2), glial fibrillary acidic protein (GFAP), and neuron cell surface antigen (A2B5) were examined 24 h and 7 days after the treatment. The results showed that both OECs‐ or B104‐CM treated AT‐MSCs express markers of progenitor and mature neurons (nestin, PGP 9.5 and MAP2) in time‐dependent manner, display morphological features resembling neuronal cells, and result negative for GFAP and A2B5, astrocyte and oligodendrocyte markers, respectively. This study demonstrated that AT‐MSCs can be influenced by the environment, indicating that these cells can respond to environmental cues also versus a neuronal phenotype. J. Cell. Physiol. 228: 2109–2118, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24386   open full text
• Targeting deregulated epigenetic control in cancer.
  Sayyed K. Zaidi, Andre J. Van Wijnen, Jane B. Lian, Janet L. Stein, Gary S. Stein.
  Journal of Cellular Physiology. July 25, 2013
  Cancer is a multifaceted disease that involves acquisition of genetic mutations, deletions, and amplifications as well as deregulation of epigenetic mechanisms that fine‐tune gene regulation. Key epigenetic mechanisms that include histone modifications, DNA methylation, and non‐coding RNA‐mediated gene silencing are often deregulated in a variety of cancers. Subnuclear localization of key proteins in the interphase nucleus and bookmarking of genes by lineage commitment factors in mitosis—a new dimension to epigenetic control of fundamental biological processes—is also modified in cancer. In this review, we discuss the various aspects of epigenetic control that are operative in a variety of cancers and their potential for risk assessment, early detection, targeted therapy, and personalized medicine. J. Cell. Physiol. 228: 2103–2108, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24387   open full text
• Caveolin and TGF‐β entanglements.
  Christoph Meyer, Yan Liu, Steven Dooley.
  Journal of Cellular Physiology. July 25, 2013
  Transforming growth factor (TGF)‐β is a multifunctional cytokine acting during development, tissue homeostasis, regeneration processes, and disease progression. Due to its pleiotropic effects, tight regulation of the induced signaling cascades is mandatory. Caveolin proteins regulate a specific endocytic pathway and modulate diverse signaling pathways and thus have been related to severe disorders, for example, cancer and fibrosis. Caveolin affects TGF‐β/‐Smad and non‐Smad signaling in many ways and thus can determine the cellular outcome upon TGF‐β challenge. Reciprocal regulation of caveolin and TGF‐β is also evident, ranging from gene expression to miRNA regulation. Finally, there is in vivo evidence that this crosstalk influences disease development and progression. This review gives an overview about the multifaceted relations of caveolin and TGF‐β. J. Cell. Physiol. 228: 2097–2102, 2013. © 2013 Wiley Periodicals, Inc.

  July 25, 2013   doi: 10.1002/jcp.24380   open full text
• MicroRNA‐195 chemosensitizes colon cancer cells to the chemotherapeutic drug doxorubicin by targeting the first binding site of BCL2L2 mRNA.
  Juan Qu, Liang Zhao, Pengzhi Zhang, Juan Wang, Ning Xu, Wenjuan Mi, Xingwang Jiang, Changming Zhang, Juan Qu.
  Journal of Cellular Physiology. March 22, 2013
  The mechanisms underlying doxorubicin (Dox) resistance in colon cancer cells are not fully understood. MicroRNA (miRNA) has emerged to play important roles in tumorigenesis and drug resistance. However, the relationship between miRNA and doxorubicin resistance in colon cancer cells has not been previously explored. In this study, we utilized microRNA array and real‐time PCR to verify that some microRNAs including miR‐127, miR‐195, miR‐22, miR‐137 were significantly down‐regulated, while miR‐21, miR‐592 were up‐regulated in both HT29/DOX and LOVO/DOX cell lines. In vitro cell viability assay showed that knockdown of miR‐195 in HT29 and LOVO cells caused a marked inhibition of Dox‐induced cytotoxicity. Moreover, we explored that miR‐195 is involved in repression of BCL2L2 expression through targeting its 3'‐untranslated region, especially the first binding site within its mRNA. Furthermore, down‐regulation of miR‐195 conferred DOX resistance in parental cells and reduced cell apoptosis activity, while over‐expression of miR‐195 sensitized resistant cells to DOX and enhanced cell apoptosis activity, all of which can be partly rescued by BCL2L2 siRNA and cDNA expression. These results may have implications for therapeutic strategies aiming to overcome colon cancer cells resistance to doxorubicin. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc.

  March 22, 2013   doi: 10.1002/jcp.24366   open full text
• Adenosine, adenosine receptors and their role in glucose homeostasis and lipid metabolism.
  Milka Koupenova, Katya Ravid.
  Journal of Cellular Physiology. March 04, 2013
  --- - |2 Abstract Adenosine is an endogenous metabolite that is released from all tissues and cells including liver, pancreas, muscle and fat, particularly under stress, intense exercise, or during cell damage. The role of adenosine in glucose homeostasis has been attributed to its ability to regulate, through its membrane receptors, processes such as insulin secretion, glucose release and clearance, glycogenolysis, and glycogenesis. Additionally, adenosine and its multiple receptors have been connected to lipid metabolism by augmenting insulin‐mediated inhibition of lipolysis, and the subsequent increase in free fatty acids and glycerol levels. Furthermore, adenosine was reported to control liver cholesterol synthesis, consequently affecting plasma levels of cholesterol and triglycerides, and the amount of fat tissue. Alterations in the balance of glucose and lipid homeostasis have implications in both cardiovascular disease and diabetes. The ability of different adenosine receptors to activate and inhibit the same signaling cascades has made it challenging to study the influence of adenosine, adenosine analogs and their receptors in health and disease. This review focuses on the role and significance of different adenosine receptors in mediating the effect of adenosine on glucose and lipid homeostasis. J. Cell. Physiol. © 2013 Wiley Periodicals, Inc. - Journal of Cellular Physiology, Volume 0, Issue ja, -Not available-.
  

  March 04, 2013   doi: 10.1002/jcp.24352   open full text