Y Chang1, S W N Ueng, S Lin-Chao, C C-K Chao. 1. Department of Biochemistry and Molecular Biology, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
Abstract
OBJECTIVE: The growth-arrest-specific protein, Gas7, has been shown to be involved in reorganization of the cytoskeleton and for inducing changes in cell shape during cell differentiation. The goals of this study were to investigate the novel role of human Gas7 (hGas7) in chondrogenic differentiation of human mesenchymal stem cells (hMSCs) and to identify the relationship between hGas7, extracellular signal-regulated kinase (ERK1/2) and SOX9 in the chondrogenic pathway. METHODS: Bone marrow-derived hMSCs were induced to undergo chondrogenic differentiation with transforming growth factor-beta1 (TGF-beta1) in an aggregate culture system. The expression of hGas7 and SOX9 and phosphorylation of ERK1/2 at multiple time points were investigated. Chondrogenic capacity was evaluated by the size of aggregates, by glycosaminoglycan content, and by type II collagen and proteoglycan deposition after interfering with expression of hGas7, ERK1/2 or SOX9. To delineate the functional role of these genes in chondrogenesis, inhibition of individual gene's expression in hMSCs, by antisense oligonucleotides or interference RNA (siRNA), and the effect on chondrogenic differentiation were also investigated. RESULTS: Treatment of hMSCs with TGF-beta1 resulted in a transient up-regulation of hGas7b, one of the hGas7 isoforms (day 3-day 5), a transient phosphorylation of ERK1/2 (0.5-4 h) and an up-regulation of SOX9 (2 h to day 14). Transient expression of hGas7b was also detected in hMSCs by reverse transcription-polymerase chain reaction at day 2 and day 3 following TGF-beta1 treatment. Interference with hGas7b production by hGas7b-specific antisense oligonucleotide or inhibition of p-ERK with PD98059, a specific inhibitor of ERK signaling pathway, or interference with SOX9 production by SOX9 siRNA all caused adverse effects of chondrogenic differentiation of hMSCs. Meanwhile, inhibition of p-ERK or SOX9 both blocked the expression of hGas7b. However, the p-ERK and SOX9 pathway was not affected by inhibition of hGas7b. CONCLUSION: These results provide evidence that the transient expression of hGas7b, regulated by activation of ERK1/2 and SOX9 pathway, is essential for chondrogenic differentiation of hMSCs.
OBJECTIVE: The growth-arrest-specific protein, Gas7, has been shown to be involved in reorganization of the cytoskeleton and for inducing changes in cell shape during cell differentiation. The goals of this study were to investigate the novel role of humanGas7 (hGas7) in chondrogenic differentiation of human mesenchymal stem cells (hMSCs) and to identify the relationship between hGas7, extracellular signal-regulated kinase (ERK1/2) and SOX9 in the chondrogenic pathway. METHODS: Bone marrow-derived hMSCs were induced to undergo chondrogenic differentiation with transforming growth factor-beta1 (TGF-beta1) in an aggregate culture system. The expression of hGas7 and SOX9 and phosphorylation of ERK1/2 at multiple time points were investigated. Chondrogenic capacity was evaluated by the size of aggregates, by glycosaminoglycan content, and by type II collagen and proteoglycan deposition after interfering with expression of hGas7, ERK1/2 or SOX9. To delineate the functional role of these genes in chondrogenesis, inhibition of individual gene's expression in hMSCs, by antisense oligonucleotides or interference RNA (siRNA), and the effect on chondrogenic differentiation were also investigated. RESULTS: Treatment of hMSCs with TGF-beta1 resulted in a transient up-regulation of hGas7b, one of the hGas7 isoforms (day 3-day 5), a transient phosphorylation of ERK1/2 (0.5-4 h) and an up-regulation of SOX9 (2 h to day 14). Transient expression of hGas7b was also detected in hMSCs by reverse transcription-polymerase chain reaction at day 2 and day 3 following TGF-beta1 treatment. Interference with hGas7b production by hGas7b-specific antisense oligonucleotide or inhibition of p-ERK with PD98059, a specific inhibitor of ERK signaling pathway, or interference with SOX9 production by SOX9 siRNA all caused adverse effects of chondrogenic differentiation of hMSCs. Meanwhile, inhibition of p-ERK or SOX9 both blocked the expression of hGas7b. However, the p-ERK and SOX9 pathway was not affected by inhibition of hGas7b. CONCLUSION: These results provide evidence that the transient expression of hGas7b, regulated by activation of ERK1/2 and SOX9 pathway, is essential for chondrogenic differentiation of hMSCs.
Authors: Kitty Chui; Alpa Trivedi; C Yan Cheng; Diana B Cherbavaz; Paul F Dazin; Ai Lam Thu Huynh; James B Mitchell; Gabriel A Rabinovich; Linda J Noble-Haeusslein; Constance M John Journal: Cell Transplant Date: 2010-11-05 Impact factor: 4.064
Authors: Z H Wang; X L Li; X J He; B J Wu; M Xu; H M Chang; X H Zhang; Z Xing; X H Jing; D M Kong; X H Kou; Y Y Yang Journal: Braz J Med Biol Res Date: 2014-03-18 Impact factor: 2.590
Authors: Leonieke M E van Koolwijk; Wishal D Ramdas; M Kamran Ikram; Nomdo M Jansonius; Francesca Pasutto; Pirro G Hysi; Stuart Macgregor; Sarah F Janssen; Alex W Hewitt; Ananth C Viswanathan; Jacoline B ten Brink; S Mohsen Hosseini; Najaf Amin; Dominiek D G Despriet; Jacqueline J M Willemse-Assink; Rogier Kramer; Fernando Rivadeneira; Maksim Struchalin; Yurii S Aulchenko; Nicole Weisschuh; Matthias Zenkel; Christian Y Mardin; Eugen Gramer; Ulrich Welge-Lüssen; Grant W Montgomery; Francis Carbonaro; Terri L Young; Céline Bellenguez; Peter McGuffin; Paul J Foster; Fotis Topouzis; Paul Mitchell; Jie Jin Wang; Tien Y Wong; Monika A Czudowska; Albert Hofman; Andre G Uitterlinden; Roger C W Wolfs; Paulus T V M de Jong; Ben A Oostra; Andrew D Paterson; David A Mackey; Arthur A B Bergen; André Reis; Christopher J Hammond; Johannes R Vingerling; Hans G Lemij; Caroline C W Klaver; Cornelia M van Duijn Journal: PLoS Genet Date: 2012-05-03 Impact factor: 5.917