Zhang Yuhan, Li Shuang, Wen Xin, Tong Huili, Li Shufeng, Yan Yunqin
The Laboratory of Cell and Development, Northeast Agricultural University, Harbin 150030, China.
Biology (Basel). 2021 Jul 20;10(7):686. doi: 10.3390/biology10070686.
Myocilin (MYOC) is a glycoprotein encoded by a gene associated with glaucoma pathology. In addition to the eyes, it also expresses at high transcription levels in the heart and skeletal muscle. MYOC affects the formation of the murine gastrocnemius muscle and is associated with the differentiation of mouse osteoblasts, but its role in the differentiation of C2C12 cells has not yet been reported. Here, MYOC expression was found to increase gradually during the differentiation of C2C12 cells. Overexpression of MYOC resulted in enhanced differentiation of C2C12 cells while its inhibition caused reduced differentiation. Furthermore, immunoprecipitation indicated that MYOC binds to Caveolin-1 (CAV1), a protein that influences the TGF-β pathway. Laser confocal microscopy also revealed the common sites of action of the two during the differentiation of C2C12 cells. Additionally, CAV1 was upregulated significantly as C2C12 cells differentiated, with CAV1 able to influence the differentiation of the cells. Furthermore, the Western blotting analysis demonstrated that the expression of MYOC affected the TGF-β pathway. Finally, MYOC was overexpressed while CAV1 was inhibited. The results indicate that reduced CAV1 expression blocked the promotion of C2C12 cell differentiation by MYOC. In conclusion, the results demonstrated that MYOC regulates TGF-β by influencing CAV1 to promote the differentiation of C2C12 cells.
肌纤蛋白(MYOC)是一种由与青光眼病理相关的基因编码的糖蛋白。除眼睛外,它在心脏和骨骼肌中也以高转录水平表达。MYOC影响小鼠腓肠肌的形成,并与小鼠成骨细胞的分化有关,但其在C2C12细胞分化中的作用尚未见报道。在此,发现MYOC在C2C12细胞分化过程中逐渐增加。MYOC的过表达导致C2C12细胞分化增强,而其抑制则导致分化减少。此外,免疫沉淀表明MYOC与小窝蛋白-1(CAV1)结合,CAV1是一种影响TGF-β信号通路的蛋白质。激光共聚焦显微镜也揭示了两者在C2C12细胞分化过程中的共同作用位点。此外,随着C2C12细胞分化,CAV1显著上调,CAV1能够影响细胞分化。此外,蛋白质印迹分析表明MYOC的表达影响TGF-β信号通路。最后,过表达MYOC同时抑制CAV1。结果表明,CAV1表达降低阻断了MYOC对C2C12细胞分化的促进作用。总之,结果表明MYOC通过影响CAV1来调节TGF-β,从而促进C2C12细胞的分化。