Yang D L, Gan M L, Tan Y, Ge G H, Li Q, Jiang Y Z, Tang G Q, Li M Z, Wang J Y, Li X W, Zhang S H, Zhu L
Farm Animal Genetics Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, 611130 China.
Sichuan Province General Station of Animal Husbandry, Chengdu, 611130 China.
Mol Biol (Mosk). 2019 Jan-Feb;53(1):44-52. doi: 10.1134/S002689841901018X.
MiR-222-3р has been implicated in tumor cell proliferation and has an important role in the differentiation and maturation of myogenic cells. However, its role in skeletal myoblast proliferation is still unclear. In this study, we found that miR-222-3р expression increases initially and then decreases during C2C12 myoblast proliferation. Using synthetic miRNA mimics and inhibitors in gain- or loss-of-function experiments, we snowed that miR-222-3р overexpression in C2C12 cells promotes myoblast proliferation and represses myofiber formation, while miR-222-3р downregulation has the opposite effect. Using a prediction program, BTG2 was identified as a possible target gene of miR-222-3р. During myogenesis, miR-222-3р mimics repress BTG2 expression, while miR-222-3р inhibitors promote BTG2 expression. Using dual-luciferase reporter assay, we further demonstrated that miR-222-3р specifically targets BTG2. Additionally, we show that siRNA-mediated downregulation of BTG2 expression in C2C12 myoblasts promotes the proliferation and suppresses differentiation. In conclusion, we provide a novel insight into the mechanism by which miR-222-3р regulates the proliferation and differentiation of C2C12 myoblasts by targeting BTG2. This information contributes to our understanding of the role of miRNAs in skeletal muscle development.
MiR-222-3p与肿瘤细胞增殖有关，并且在成肌细胞的分化和成熟过程中发挥重要作用。然而，其在骨骼肌成肌细胞增殖中的作用仍不清楚。在本研究中，我们发现MiR-222-3p的表达在C2C12成肌细胞增殖过程中先升高后降低。在功能获得或缺失实验中使用合成的miRNA模拟物和抑制剂，我们发现C2C12细胞中MiR-222-3p的过表达促进成肌细胞增殖并抑制肌纤维形成，而MiR-222-3p的下调则产生相反的效果。使用预测程序，BTG2被鉴定为MiR-222-3p的一个可能靶基因。在肌生成过程中，MiR-222-3p模拟物抑制BTG2表达，而MiR-222-3p抑制剂促进BTG2表达。使用双荧光素酶报告基因检测，我们进一步证明MiR-222-3p特异性靶向BTG2。此外，我们表明，C2C12成肌细胞中siRNA介导的BTG2表达下调促进增殖并抑制分化。总之，我们为MiR-222-3p通过靶向BTG2调节C2C12成肌细胞增殖和分化的机制提供了新的见解。这些信息有助于我们理解miRNA在骨骼肌发育中的作用。
