Most recent papers:

• 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