Role of Endogenous TGF-β Family in Myogenic Differentiation of C2C12 Cells
Role of Endogenous TGF-β Family in Myogenic Differentiation of C2C12 Cells
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DOI：
10.1002/jcb.22953
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发表时间：
2011-02-01
影响因子：
4
通讯作者：
Funaba, Masayuki
中科院分区：
文献类型：
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作者：
Furutani, Yuuma;Umemoto, Takenao;Funaba, Masayuki
本研究评估了BMP和tgf - β家族代表成员tgf - β在C2C12细胞肌管分化过程中的内源性活性和作用。由于tgf - β家族成员以配体依赖的方式通过Smad的磷酸化发出信号，因此监测了c端丝氨酸的Smad磷酸化。作为BMP活性指标的磷酸化Smad1/5/8在分化前表达较高，分化刺激后表达迅速下降。分化相关的变化与众所周知的bmp应答基因Ids的表达一致。在C2C12成肌细胞中使用BMP 1型受体抑制剂或noggin抑制剂可下调Myf5和MyoD等成肌调节因子的表达，导致肌管形成受损。在成肌细胞期加入BMP-2也通过下调Myf5和MyoD抑制肌管分化。与内源性BMP活性相比，Smad2（一种对tgf - β有反应的Smad）在分化刺激后8-16天的磷酸化水平更高。tgf - β I型受体抑制剂A-83-01增加Myf5和MyoD的表达，促进肌管形成。目前的研究结果表明，tgf - β家族的内源性活性在肌肉形成过程中以一种途径特异性的方式发生改变，并且这些活性是肌肉形成所必需的。j .细胞。中国生物医学工程学报，2011。(C) 2010 Wiley-Liss, Inc。
The present study evaluated endogenous activities and the role of BMP and transforming growth factor-beta (TGF-beta), representative members of the TGF-beta family, during myotube differentiation in C2C12 cells. Smad phosphorylation at the C-terminal serines was monitored, since TGF-beta family members signal via the phosphorylation of Smads in a ligand-dependent manner. Expression of phosphorylated Smad1/5/8, which is an indicator of BMP activity, was higher before differentiation, and rapidly decreased after differentiation stimulation. Differentiation-related changes were consistent with those in the expression of Ids, well-known BMP-responsive genes. Treatment with inhibitors of BMP type 1 receptors or noggin in C2C12 myoblasts down-regulated the expression of myogenic regulatory factors, such as Myf5 and MyoD, leading to impaired myotube formation. Addition of BMP-2 during the myoblast phase also inhibited myotube differentiation through the down-regulation of Myf5 and MyoD. In contrast to endogenous BMP activity, the phosphorylation of Smad2, a TGF-beta-responsive Smad, was higher 8-16 days after differentiation stimulation. A-83-01, an inhibitor of TGF-beta type I receptor, increased the expression of Myf5 and MyoD, and enhanced myotube formation. The present results reveal that endogenous activities of the TGF-beta family are changed during myogenesis in a pathway-specific manner, and that the activities are required for myogenesis. J. Cell. Biochem. 112: 614-624, 2011. (C) 2010 Wiley-Liss, Inc.