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MicroRNA-129-5p inhibits C2C12 myogenesis and represses slow fiber gene expression in vitro

机译:microRNA-129-5P抑制C2C12肌生成,体外抑制慢纤维基因表达

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The miR-129 family is widely reported as tumor repressors, although their roles in skeletal muscle have not been fully investigated. Here, the function and mechanism of miR-129-5p in skeletal muscle, a member of the miR-129 family, were explored using C2C12 cell line. Our study showed that miR-129-5p was widely detected in mouse tissues, with the highest expression in skeletal muscle. Gain- and loss-of-function study showed that miR-129-5p could negatively regulate myogenic differentiation, indicated by reduced ratio of MyHC-positive myofibers and repressed expression of myogenic genes, such as MyoD, MyoG, and MyHC. Furthermore, miR-129-5p was more enriched in fast extensor digitorum longus (EDL) than in slow soleus (SOL). Enhanced miR-129-5p could significantly reduce the expression of mitochondrial cox family, together with that of MyHC I, and knockdown of miR-129-5p conversely increased the expression of cox genes and MyHC I. Mechanistically, miR-129-5p directly targeted the 3'-UTR of Mef2a, which was suppressed by miR-129-5p agomir at both mRNA and protein levels in C2C12 cells. Moreover, overex-pression of Mef2a could rescue the inhibitory effects of miR-129-5p on the expression of myogenic factors and MyHC I. Collectively, our data revealed that miR-129-5p is a negative regulator of myogenic differentiation and slow fiber gene expression, thus affecting body metabolic homeostasis.
机译:miR-129家族被广泛报道为肿瘤抑制因子,尽管它们在骨骼肌中的作用尚未得到充分研究。在这里,我们利用C2C12细胞系探讨了miR-129家族成员骨骼肌中miR-129-5p的功能和机制。我们的研究表明,miR-129-5p广泛存在于小鼠组织中,在骨骼肌中表达最高。功能获得和功能丧失研究表明,miR-129-5p可负性调节肌源性分化,这表现为MyHC阳性肌纤维比例降低,肌源性基因如MyoD、MyoG和MyHC的表达受到抑制。此外,miR-129-5p在快速趾长伸肌(EDL)中比在慢速比目鱼肌(SOL)中更丰富。增强的miR-129-5p可以显著降低线粒体cox家族和MyHC I的表达,而敲除miR-129-5p则相反地增加cox基因和MyHC I的表达。从机制上讲,miR-129-5p直接靶向Mef2a的3'-UTR,在C2C12细胞的mRNA和蛋白质水平上,它都被miR-129-5p agomir抑制。此外,过度表达Mef2a可以挽救miR-129-5p对肌源性因子和MyHC I表达的抑制作用。总体而言,我们的数据显示,miR-129-5p是肌源性分化和慢纤维基因表达的负调节因子,从而影响机体代谢稳态。

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