bluuebunny/crossref_metadata_2025_split huggingface dataset:
The transforming growth factor‐beta (TGF‐β) mediates acquisition of a mesenchymal stem cell‐like phenotype in human liver cells
Authors: Caja Laia , Bertran Esther , Campbell Jean , Fausto Nelson , Fabregat Isabel 
Published: October 2010

AbstractTransforming growth factor‐beta (TGF‐β) mediates several and sometime opposite effects in epithelial cells, inducing growth inhibition, and apoptosis but also promoting an epithelial to mesenchymal transition (EMT) process, which enhances cell migration and invasion. TGF‐β plays relevant roles in different liver pathologies; however, very few is known about its specific signaling and cellular effects in human primary hepatocytes. Here we show that TGF‐β inhibits proliferation and induces pro‐apoptotic genes (such as BMF or BIM) in primary cultures of human fetal hepatocytes (HFH), but also up‐regulates anti‐apoptotic genes, such as BCL‐XL and XIAP. Inhibition of the epidermal growth factor receptor (EGFR), using gefitinib, abrogates the increase in the expression of the anti‐apoptotic genes and significantly enhances cell death. Simultaneously, TGF‐β is able to induce an EMT process in HFH, coincident with Snail up‐regulation and a decrease in E‐cadherin levels, cells showing mesenchymal proteins and reorganization of the actin cytoskeleton in stress fibers. Interestingly, these cells show loss of expression of specific hepatic genes and increased expression of stem cell markers. Chronic treatment with TGF‐β allows selection of a population of mesenchymal cells with a de‐differentiated phenotype, reminiscent of progenitor‐like cells. Process is reversible and the mesenchymal stem‐like cells re‐differentiate to hepatocytes under controlled experimental conditions. In summary, we show for the first time that human hepatocytes may respond to TGF‐β inducing different signals, some of them might contribute to tumor suppression (growth inhibition and apoptosis), but others should mediate liver tumor progression and invasion (EMT and acquisition of a stem‐like phenotype). J. Cell. Physiol. 226: 1214–1223, 2011. © 2010 Wiley‐Liss, Inc.
Fibroblast cytoskeletal remodeling contributes to connective tissue tension
Authors: Langevin Helene M. , Bouffard Nicole A. , Fox James R. , Palmer Bradley M. , Wu Junru , Iatridis James C. , Barnes William D. , Badger Gary J. , Howe Alan K. 
Published: October 2010

AbstractThe visco‐elastic behavior of connective tissue is generally attributed to the material properties of the extracellular matrix rather than cellular activity. We have previously shown that fibroblasts within areolar connective tissue exhibit dynamic cytoskeletal remodeling within minutes in response to tissue stretch ex vivo and in vivo. Here, we tested the hypothesis that fibroblasts, through this cytoskeletal remodeling, actively contribute to the visco‐elastic behavior of the whole tissue. We measured significantly increased tissue tension when cellular function was broadly inhibited by sodium azide and when cytoskeletal dynamics were compromised by disrupting microtubules (with colchicine) or actomyosin contractility (via Rho kinase inhibition). These treatments led to a decrease in cell body cross‐sectional area and cell field perimeter (obtained by joining the end of all of a fibroblast's processes). Suppressing lamellipodia formation by inhibiting Rac‐1 decreased cell body cross‐sectional area but did not affect cell field perimeter or tissue tension. Thus, by changing shape, fibroblasts can dynamically modulate the visco‐elastic behavior of areolar connective tissue through Rho‐dependent cytoskeletal mechanisms. These results have broad implications for our understanding of the dynamic interplay of forces between fibroblasts and their surrounding matrix, as well as for the neural, vascular, and immune cell populations residing within connective tissue. J. Cell. Physiol. 226: 1166–1175, 2011. © 2010 Wiley‐Liss, Inc.
Identification of heat shock protein 5, calnexin and integral membrane protein 2B as Adam7‐interacting membrane proteins in mouse sperm
Authors: Han Cecil , Park Inju , Lee Boyeon , Jin Sora , Choi Heejin , Kwon Jun Tae , Kwon Yong‐il , Kim Do Han , Park Zee Yong , Cho Chunghee 
Published: October 2010

AbstractIn mammals, sperm acquire their motility and ability to fertilize eggs in the epididymis. This maturation process involves the acquisition of particular proteins from the epididymis. One such secretory protein specifically expressed in the epididymis is Adam7 (a disintegrin and metalloprotease 7). Previous studies have shown that Adam7 that resides in an intracellular compartment of epididymal cells is transferred to sperm membranes, where its levels are dependent on the expression of Adam2 and Adam3, which have critical roles in fertilization. Here, using a proteomics approach based on mass spectrometry, we identified proteins that interact with Adam7 in sperm membranes. This analysis revealed that Adam7 forms complexes with calnexin (Canx), heat shock protein 5 (Hspa5), and integral membrane protein 2B (Itm2b). Canx and Hspa5 are molecular chaperones, and Itm2b is a type II integral membrane protein implicated in neurodegeneration. The interaction of Adam7 with these proteins was confirmed by immunoprecipitation‐Western blot analysis. We found that Adam7 and Itm2b are located in detergent‐resistant regions known to be highly correlated with membrane lipid rafts. We further found that the association of Adam7 with Itm2b is remarkably promoted during sperm capacitation owing to a conformational change of Adam7 that occurs in concert with the capacitation process. Thus, our results suggest that Adam7 functions in fertilization through the formation of a chaperone complex and enhanced association with Itm2b during capacitation in sperm. J. Cell. Physiol. 226: 1186–1195, 2011. © 2010 Wiley‐Liss, Inc.
CXCL5/ENA78 increased cell migration and epithelial‐to‐mesenchymal transition of hormone‐independent prostate cancer by early growth response‐1/snail signaling pathway
Authors: Kuo Po‐Lin , Chen Yen‐Hsu , Chen Tun‐Chieh , Shen Kun‐Hung , Hsu Ya‐Ling 
Published: October 2010

AbstractProstate cancers that are resistant to hormone therapy are more invasive and have greater ability to spread to other organs than androgen‐dependent prostate cancers. Furthermore, this type of prostate cancer is also highly resistant to current forms of chemotherapy. This study analyzed CXCL5/ENA78, which is highly expressed in androgen‐independent prostate cancers, and is responsible for cell migration and epithelial‐to‐mesenchymal transition in two androgen‐independent prostate cancer cell lines. Inducement of PC‐3 and DU145 cancer progression by CXCL5/ENA78 is associated with increased Raf/MEK/ERK activation, and the upregulation of early growth response‐1 (Egr‐1) and Snail. Blockade of Egr‐1 decreased Snail upregulation and cell migration, indicating that Egr‐1 is required in CXCL5/ENA78‐mediated Snail enhancement and cell migration. In addition, Egr‐1 siRNA also decreased the effect of CXCL5/ENA78 on p27 inhibition, Cdk4 induction and cell proliferation, suggesting Egr‐1 is also involved in CXCL5/ENA78‐mediated cell growth. Moreover, blocking ERK1/2 by siRNA suppressed CXCL5/ENA78‐induced Egr‐1 enhancement, cell migration, and proliferation. Our study suggests that inhibition of CXCL5/ENA78‐mediated ERK/Egr‐1/Snail signaling is an attractive therapeutic target for androgen‐independent prostate cancer. J. Cell. Physiol. 226: 1224–1231, 2011. © 2010 Wiley‐Liss, Inc.
Differential role of Rho GTPases in intestinal epithelial barrier regulation in vitro
Authors: Schlegel Nicolas , Meir Michael , Spindler Volker , Germer Christoph‐Thomas , Waschke Jens 
Published: October 2010

AbstractMaintenance of intestinal epithelial barrier functions is crucial to prevent systemic contamination by microbes that penetrate from the gut lumen. GTPases of the Rho‐family such as RhoA, Rac1, and Cdc42 are known to be critically involved in the regulation of intestinal epithelial barrier functions. However, it is still unclear whether inactivation or activation of these GTPases exerts barrier protection or not. We tested the effects of Rho GTPase activities on intestinal epithelial barrier functions by using the bacterial toxins cytotoxic necrotizing factor 1 (CNF‐1), toxin B, C3 transferase (C3 TF), and lethal toxin (LT) in an in vitro model of the intestinal epithelial barrier. Incubation of cell monolayers with CNF‐1 for 3 h induced exclusive activation of RhoA whereas Rac1 and Cdc42 activities were unchanged. As revealed by FITC‐dextran flux and measurements of transepithelial electrical resistance (TER) intestinal epithelial permeability was significantly increased under these conditions. Inhibition of Rho kinase via Y27632 blocked barrier destabilization of CNF‐1 after 3 h. In contrast, after 24 h of incubation with CNF‐1 only Rac1 and Cdc42 but not RhoA were activated which resulted in intestinal epithelial barrier stabilization. Toxin B to inactivate RhoA, Rac1, and Cdc42 as well as Rac1 inhibitor LT increased intestinal epithelial permeability. Similar effects were observed after inhibition of RhoA/Rho kinase signaling by C3 TF or Y27632. Taken together, these data demonstrate that both activation and inactivation of RhoA signaling increased paracellular permeability whereas activation of Rac1 and Cdc42 correlated with stabilized barrier functions. J. Cell. Physiol. 226: 1196–1203, 2011. © 2010 Wiley‐Liss, Inc.
Substance P stimulates the recovery of bone marrow after the irradiation
Authors: An You Sun , Lee EunAh , Kang Mi‐Hyun , Hong Hyun Sook , Kim Mi‐Ra , Jang Won Seok , Son Youngsook , Yi Jae Youn 
Published: October 2010

AbstractThe therapeutic use of ionizing radiation (e.g., X‐rays and γ‐rays) needs to inflict minimal damage on non‐target tissue. Recent studies have shown that substance P (SP) mediates multiple activities in various cell types, including cell proliferation, anti‐apoptotic responses, and inflammatory processes. The present study investigated the effects of SP on γ‐irradiated bone marrow stem cells (BMSCs). In mouse bone marrow extracts, SP prolonged activation of Erk1/2 and enhanced Bcl‐2 expression, but attenuated the activation of apoptotic molecules (e.g., p38 and cleaved caspase‐3) and down‐regulated Bax. We also observed that SP‐decreased apoptotic cell death and stimulated cell proliferation in γ‐irradiated mouse bone marrow tissues through TUNEL assay and PCNA analysis. To determine how SP affects bone marrow stem cell populations, mouse bone marrow cells were isolated and colony‐forming unit (CFU) of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) was estimated. SP‐pretreated ones showed higher CFUs of MSC and HSC than untreated ones. Furthermore, when SP was pretreated in cultured human MSC, it significantly decreased apoptotic cells at 48 and 72 h after γ‐irradiation. Compared with untreated cells, SP‐treated human MSCs showed reduced cleavage of apoptotic molecules such as caspase‐8, ‐9, ‐3, and poly ADP‐ribose polymerase (PARP). Thus, our results suggest that SP alleviates γ‐radiation‐induced damage to mouse BMSCs and human MSCs via regulation of the apoptotic pathway. J. Cell. Physiol. 226: 1204–1213, 2011. © 2010 Wiley‐Liss, Inc.
Role of Smad2/3 and p38 MAP kinase in TGF‐β1‐induced epithelial–mesenchymal transition of pulmonary epithelial cells
Authors: Kolosova Irina , Nethery David , Kern Jeffrey A. 
Published: October 2010

AbstractIdiopathic pulmonary fibrosis is characterized by myofibroblast accumulation, extracellular matrix (ECM) remodeling, and excessive collagen deposition. ECM‐producing myofibroblasts may originate from epithelial cells through epithelial to mesenchymal transition (EMT). TGF‐β1 is an inducer of EMT in pulmonary epithelial cells in vitro and in vivo, though the mechanisms are unclear. We hypothesized that TGF‐β1 induced EMT through Smad‐dependent and ‐independent processes. To test this hypothesis, we studied the roles and mechanisms of TGF‐β1‐induced Smad and p38 mitogen‐activated protein kinase (MAPK) signaling in EMT‐related changes in pulmonary epithelial cells. Exposure of pulmonary epithelial 1HAEo− cells to TGF‐β1 resulted in morphological and molecular changes of EMT over a 96‐h period; loss of cell–cell contact, cell elongation, down‐regulation of E‐cadherin, up‐regulation of fibronectin, and up‐regulation of collagen I. Both Smad2/3 and p38 MAPK signaling pathways were activated by TGF‐β1. However, neither Smad2/3 nor p38 MAPK were required for the down‐regulation of E‐cadherin, yet p38 MAPK was associated with fibronectin up‐regulation. Both Smad2/3 and p38 MAPK had a role in regulation of TGF‐β1‐induced collagen expression. Furthermore, these data demonstrate that Smads and p38 MAPK differentially regulate EMT‐related changes in pulmonary epithelial cells. J. Cell. Physiol. 226: 1248–1254, 2011. © 2010 Wiley‐Liss, Inc.
The inhibitory effect of ginsan on TGF‐β mediated fibrotic process
Authors: Ahn Ji‐Yeon , Kim Mi‐Hyoung , Lim Min‐Jin , Park Sarah , Lee Sae‐lo‐oom , Yun Yeon‐Sook , Song Jie‐Young 
Published: October 2010

AbstractTransforming growth factor‐beta (TGF‐β) plays a central role in the development of fibrosis by stimulating extracellular matrix accumulation, and signals either directly or indirectly through types I, II, and III (TβRI, II, and III) TGF‐β receptor complexes. Ginsan, a polysaccharide extracted from Panax ginseng, has multiple immunomodulatory effects. Here, we examine whether ginsan regulates the fibrogenic process by interfering with TGF‐β signaling pathways. TGF‐β treatment of murine or human normal lung fibroblasts enhanced the levels of several fibrotic markers, including smooth muscle alpha actin (α‐SMA), collagen‐1, and fibronectin. Interestingly, ginsan treatment either before or after TGF‐β administration led to significant reductions in all of α‐SMA, collagen‐1, and fibronectin expression levels. Ginsan not only inhibited phosphorylation of Smad2 and Smad3, but also attenuated pERK and pAKT signaling induced by TGF‐β. Moreover, ginsan restored TβRIII protein expression, which was significantly downregulated by TGF‐β, but reduced TβRI and TβRII protein levels. In a murine model of bleomycin (BLM)‐induced pulmonary fibrosis, ginsan significantly suppressed accumulation of collagen, α‐SMA, and TGF‐β. These data collectively suggest that ginsan acts as an effective anti‐fibrotic agent in the treatment of pulmonary fibrosis by blocking multiple TGF‐β signaling pathways. J. Cell. Physiol. 226: 1241–1247, 2011. © 2010 Wiley‐Liss, Inc.
Leptin increases motility and integrin up‐regulation in human prostate cancer cells
Authors: Huang Chih‐Yang , Yu Hsin‐Shan , Lai Tung‐Yuan , Yeh Yu‐Lan , Su Cheng‐Chuan , Hsu His‐Hsien , Tsai Fuu‐Jen , Tsai Chang‐Hai , Wu Hsi‐Chin , Tang Chih‐Hsin 
Published: October 2010

AbstractProstate cancer is the most commonly diagnosed malignancy in men and shows a predilection for metastasis to distant organs. Leptin, an adipocyte‐derived cytokine that is closely associated with obesity, has recently been shown to be involved in carcinogenesis and cancer progression. The aim of this study was to investigate whether leptin is associated with the motility of prostate cancer cells. We found that leptin increased the migration of human prostate cancer cells and expression of αvβ3 integrin on these cells. Leptin‐mediated migration and increased integrin expression were attenuated by OBRl receptor antisense oligonucleotide (ODN). Activation of insulin receptor substrate (IRS‐1), phosphatidylinositol 3‐kinase (PI3K), Akt, and NF‐κB pathways after leptin treatment was demonstrated. Furthermore, leptin‐induced integrin expression and migration activity were inhibited by specific inhibitors; small interfering RNAs (siRNAs); and mutants of the IRS‐1, PI3K, Akt, and NF‐κB cascades. Therefore, this study shows that leptin stimulates the migration of human prostate cancer cells, one of the mechanisms underlying leptin‐directed migration was transcriptional up‐regulation of αvβ3 integrin expression through the OBR1/IRS‐1/PI3K/Akt/NF‐κB signal transduction pathway. J. Cell. Physiol. 226: 1274–1282, 2011. © 2010 Wiley‐Liss, Inc.
Neutrophil differentiation from human‐induced pluripotent stem cells
Authors: Morishima Tatsuya , Watanabe Ken‐ichiro , Niwa Akira , Fujino Hisanori , Matsubara Hiroshi , Adachi Souichi , Suemori Hirofumi , Nakahata Tatsutoshi , Heike Toshio 
Published: October 2010

AbstractInduced pluripotent stem (iPS) cells are of potential value not only for regenerative medicine, but also for disease investigation. The present study describes the development of a neutrophil differentiation system from human iPS cells (hiPSCs) and the analysis of neutrophil function and differentiation. The culture system used consisted of the transfer of hiPSCs onto OP9 cells and their culture with vascular endothelial growth factor (VEGF). After 10 days, TRA 1‐85+CD34+VEGF receptor‐2 (VEGFR‐2)high cells were sorted and co‐cultured with OP9 cells in the presence of hematopoietic cytokines for 30 days. Floating cells were collected and subjected to morphological and functional analysis. These hiPSC‐derived neutrophils were similar to peripheral blood mature neutrophils in morphology, contained functional neutrophil specific granules, and were equipped with the basic functions such as phagocytosis, superoxide production, and chemotaxis. In the process of differentiation, myeloid cells appeared sequentially from immature myeloblasts to mature segmented neutrophils. Expression patterns of surface antigen, transcription factors, and granule proteins during differentiation were also similar to those of granulopoiesis in normal bone marrow. In conclusion, differentiation of mature neutrophils from hiPSCs was successfully induced in a similar process to normal granulopoiesis using an OP9 co‐culture system. This system may be applied to elucidate the pathogenesis of various hematological diseases that affect neutrophils. J. Cell. Physiol. 226: 1283–1291, 2011. © 2010 Wiley‐Liss, Inc.
Retinoic acid receptor‐mediated signaling protects cardiomyocytes from hyperglycemia induced apoptosis: Role of the renin‐angiotensin system
Authors: Guleria Rakeshwar S. , Choudhary Rashmi , Tanaka Takemi , Baker Kenneth M. , Pan Jing 
Published: October 2010

AbstractDiabetes mellitus (DM) is a primary risk factor for cardiovascular diseases and heart failure. Activation of the retinoic acid receptor (RAR) and retinoid X receptor (RXR) has an anti‐diabetic effect; but, a role in diabetic cardiomyopathy remains unclear. Using neonatal and adult cardiomyocytes, we determined the role of RAR and RXR in hyperglycemia‐induced apoptosis and expression of renin‐angiotensin system (RAS) components. Decreased nuclear expression of RARα and RXRα, activation of apoptotic signaling and cell apoptosis was observed in high glucose (HG) treated neonatal and adult cardiomyocytes and diabetic hearts in Zucker diabetic fatty (ZDF) rats. HG‐induced apoptosis and reactive oxygen species (ROS) generation was prevented by both RAR and RXR agonists. Silencing expression of RARα and RXRα, by small interference RNA, promoted apoptosis under normal conditions and significantly enhanced HG‐induced apoptosis, indicating that RARα and RXRα are required in regulating cell apoptotic signaling. Blocking angiotensin type 1 receptor (AT1R); but, not AT2R, attenuated HG‐induced apoptosis and ROS generation. Moreover, HG induced gene expression of angiotensinogen, renin, AT1R, and angiotensin II (Ang II) synthesis were inhibited by RARα agonists and promoted by silencing RARα. Activation of RXRα, downregulated the expression of AT1R; and RXRα silencing accelerated HG induced expression of angiotensinogen and Ang II synthesis, whereas there was no significant effect on renin gene expression. These results indicate that reduction in the expression of RARα and RXRα has an important role in hyperglycemia mediated apoptosis and expression of RAS components. Activation of RAR/RXR signaling protects cardiomyocytes from hyperglycemia, by reducing oxidative stress and inhibition of the RAS. J. Cell. Physiol. 226: 1292–1307, 2011. © 2010 Wiley‐Liss, Inc.
Gene profiling reveals hydrogen sulphide recruits death signaling via the N‐methyl‐D‐aspartate receptor identifying commonalities with excitotoxicity
Authors: Chen Minghui Jessica , Peng Zhao Feng , Manikandan Jayapal , Melendez Alirio J. , Tan Gek San , Chung Ching Ming , Li Qiu‐Tian , Tan Theresa M. , Deng Lih Wen , Whiteman Matthew , Beart Philip M. , Moore Phillip K. , Cheung Nam Sang 
Published: October 2010

AbstractRecently the role of hydrogen sulphide (H2S) as a gasotransmitter stimulated wide interest owing to its involvement in Alzheimer's disease and ischemic stroke. Previously we demonstrated the importance of functional ionotropic glutamate receptors (GluRs) by neurons is critical for H2S‐mediated dose‐ and time‐dependent injury. Moreover N‐methyl‐D‐aspartate receptor (NMDAR) antagonists abolished the consequences of H2S‐induced neuronal death. This study focuses on deciphering the downstream effects activation of NMDAR on H2S‐mediated neuronal injury by analyzing the time‐course of global gene profiling (5, 15, and 24 h) to provide a comprehensive description of the recruitment of NMDAR‐mediated signaling. Microarray analyses were performed on RNA from cultured mouse primary cortical neurons treated with 200 µM sodium hydrosulphide (NaHS) or NMDA over a time‐course of 5–24 h. Data were validated via real‐time PCR, western blotting, and global proteomic analysis. A substantial overlap of 1649 genes, accounting for over 80% of NMDA global gene profile present in that of H2S and over 50% vice versa, was observed. Within these commonly occurring genes, the percentage of transcriptional consistency at each time‐point ranged from 81 to 97%. Gene families involved included those related to cell death, endoplasmic reticulum stress, calcium homeostasis, cell cycle, heat shock proteins, and chaperones. Examination of genes exclusive to H2S‐mediated injury (43%) revealed extensive dysfunction of the ubiquitin‐proteasome system. These data form a foundation for the development of screening platforms and define targets for intervention in H2S neuropathologies where NMDAR‐activated signaling cascades played a substantial role. J. Cell. Physiol. 226: 1308–1322, 2011. © 2010 Wiley‐Liss, Inc.
G‐protein coupled receptor kinase 5 mediates lipopolysaccharide‐induced NFκB activation in primary macrophages and modulates inflammation in vivo in mice
Authors: Patial Sonika , Shahi Shipra , Saini Yogesh , Lee Taehyung , Packiriswamy Nandakumar , Appledorn Daniel M. , LaPres John J. , Amalfitano Andrea , Parameswaran Narayanan 
Published: October 2010

AbstractG‐protein coupled receptor kinase‐5 (GRK5) is a serine/threonine kinase discovered for its role in the regulation of G‐protein coupled receptor signaling. Recent studies have shown that GRK5 is also an important regulator of signaling pathways stimulated by non‐GPCRs. This study was undertaken to determine the physiological role of GRK5 in Toll‐like receptor‐4‐induced inflammatory signaling pathways in vivo and in vitro. Using mice genetically deficient in GRK5 (GRK5−/−) we demonstrate here that GRK5 is an important positive regulator of lipopolysaccharide (LPS, a TLR4 agonist)‐induced inflammatory cytokine and chemokine production in vivo. Consistent with this role, LPS‐induced neutrophil infiltration in the lungs (assessed by myeloperoxidase activity) was markedly attenuated in the GRK5−/− mice compared to the GRK5+/+ mice. Similar to the in vivo studies, primary macrophages from GRK5−/− mice showed attenuated cytokine production in response to LPS. Our results also identify TLR4‐induced NFκB pathway in macrophages to be selectively regulated by GRK5. LPS‐induced IκBα phosphorylation, NFκB p65 nuclear translocation, and NFκB binding were markedly attenuated in GRK5−/− macrophages. Together, our findings demonstrate that GRK5 is a positive regulator of TLR4‐induced IκBα–NFκB pathway as well as a key modulator of LPS‐induced inflammatory response. J. Cell. Physiol. 226: 1323–1333, 2011. © 2010 Wiley‐Liss, Inc.