miR-378a-3p promotes differentiation and inhibits proliferation of myoblasts by targeting HDAC4 in skeletal muscle development
miR-378a-3p promotes differentiation and inhibits proliferation of myoblasts by targeting HDAC4 in skeletal muscle development
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miR-378a-3p通过靶向骨骼肌发育中的HDAC4促进成肌细胞分化并抑制增殖
DOI：
10.1080/15476286.2016.1239008
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发表时间：
2016-01-01
期刊：
影响因子：
4.1
通讯作者：
Chen, Hong
中科院分区：
文献类型：
--
作者：
Wei, Xuefeng;Li, Hui;Chen, Hong
摘要肌肉发育，或肌肉发生，是一个高度调控的复杂过程。MicroRNAs的一个子集(MiRNAs)已被确定为肌肉发生的关键调节因子。最近，miR-378a被发现参与肌肉发生，但miR-378a如何调控成肌细胞的增殖和分化的机制尚未确定。我们发现miR-378a-3p在肌肉中的表达显著高于在其他组织中的表达，提示miR-378a-3p在肌肉发育中起重要作用。MiR-378a-3p过表达后，C2C12成肌细胞中MyoD和MHC的表达均在mRNA和蛋白水平上增加，证实miR-378a-3p促进了肌细胞的分化。CCK-8检测和Annexin V-FITC/PI染色结果表明，强制表达miR-378a-3p可促进C2C12细胞的凋亡。经TargetScan鉴定，组蛋白乙酰化酶4(HDAC4)是miR-378a-3p的潜在靶点。我们用双荧光素酶实验和Western blotting证实了miR-378a-3p靶向HDAC4。我们的RNAi分析结果还表明，HDAC4显著促进C2C12细胞的分化，并通过Bcl2抑制细胞存活。因此，我们得出结论，miR-378a-3p通过转录后下调HDAC4来调节骨骼肌生长并促进成肌细胞的分化。
ABSTRACT Muscle development, or myogenesis, is a highly regulated, complex process. A subset of microRNAs (miRNAs) have been identified as critical regulators of myogenesis. Recently, miR-378a was found to be involved in myogenesis, but the mechanism of how miR-378a regulates the proliferation and differentiation of myoblasts has not been determined. We found that miR-378a-3p expression in muscle was significantly higher than in other tissues, suggesting an important effect on muscle development. Overexpression of miR-378a-3p increased the expression of MyoD and MHC in C2C12 myoblasts both at the level of mRNA and protein, confirming that miR-378a-3p promoted muscle cell differentiation. The forced expression of miR-378a-3p promoted apoptosis of C2C12 cells as evidenced by CCK-8 assay and Annexin V-FITC/PI staining results. Through TargetScan, histone acetylation enzyme 4 (HDAC4) was identified as a potential target of miR-378a-3p. We confirmed targeting of HDAC4 by miR-378a-3p using a dual luciferase assay and western blotting. Our RNAi analysis results also showed that HDAC4 significantly promoted differentiation of C2C12 cells and inhibited cell survival through Bcl-2. Therefore, we conclude that miR-378a-3p regulates skeletal muscle growth and promotes the differentiation of myoblasts through the post-transcriptional down-regulation of HDAC4.