Most recent papers:

• 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
• 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. September 08, 2017
  Background 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. Methods and results 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. Conclusions Together, these results reveal that excess ROS generation is involved in the LPS‐induced defects in heart tube during chick embryo development. This article is protected by copyright. All rights reserved

  September 08, 2017   doi: 10.1002/jcp.26175   open full text
• Novel role for the testis‐enriched HSPA2 protein in regulating epidermal keratinocyte differentiation.
  Agnieszka Gogler‐Pigłowska, Katarzyna Klarzyńska, Damian R. Sojka, Anna Habryka, Magdalena Głowala‐Kosińska, Marcin Herok, Mariusz Kryj, Monika Halczok, Zdzisław Krawczyk, Dorota Scieglinska.
  Journal of Cellular Physiology. September 07, 2017
  HSPA2, a poorly characterized member of the HSPA (HSP70) chaperone family, is a testis‐enriched protein involved in male germ cell differentiation. Previously, we revealed that HSPA2 is present in human stratified epithelia, including epidermis, however the contribution of this protein to epithelial biology remained unknown. Here, we show for the first time that HSPA2 is expressed in basal epidermal keratinocytes, albeit not in keratinocytes exhibiting features attributed to primitive undifferentiated progenitors, and participates in the keratinocyte differentiation process. We found that HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. We also shown that lentiviral‐mediated shRNA silencing of HSPA2 expression in HaCaT cells caused a set of phenotypic changes characteristic for keratinocytes committed to terminal differentiation such as reduced clonogenic potential, impaired adhesiveness and increased basal and confluency‐induced expression of differentiation markers. Moreover, the fraction of undifferentiated cells that rapidly adhered to collagen IV was less numerous in HSPA2‐deficient cells than in the control. In a 3D reconstructed human epidermis model, HSPA2 deficiency resulted in accelerated development of a filaggrin‐positive layer. Collectively, our results clearly show a link between HSPA2 expression and maintenance of keratinocytes in an undifferentiated state in the basal layer of the epidermis. It seems that HSPA2 could retain keratinocytes from premature entry into the terminal differentiation process. Overall, HSPA2 appears to be necessary for controlling development of properly stratified epidermis and thus for maintenance of skin homeostasis. In human skin HSPA2 is preferentially expressed in basal epidermal keratinocytes. HSPA2 is dispensable for protection of HaCaT keratinocytes against heat shock‐induced cytotoxicity. HSPA2 participates in the keratinocyte differentiation process.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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