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首页> 外文期刊>Journal of genetics >MiR-27b promotes sheep skeletal muscle satellite cell proliferation by targeting myostatin gene
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MiR-27b promotes sheep skeletal muscle satellite cell proliferation by targeting myostatin gene

机译：MiR-27b通过靶向myostatin基因促进绵羊​​骨骼肌卫星细胞增殖

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To investigate the role of miR-27b in sheep skeletal muscle development, here we first cloned the sequence of sheep pre-miR-27b, then further investigated its expression pattern in sheep skeletal muscle in vivo, the relationship of miR-27b expression and sheep skeletal muscle satellite cell proliferation and differentiation in vitro, and then finally confirmed its target gene during this development process. MiR-27b sequence, especially its mature sequence, was conservative among different species. MiR-27b highly expressed in sheep skeletal muscle than other tissues. In skeletal muscle of Suffolk and Bashbay sheep, miR-27b was upregulated during foetal period and downregulated during postnatal period significantly (P0.01), but it still kept a relatively higher expressionlevel in skeletal muscle of postnatal Suffolk sheep than Bashbay. There is a potential target site of miR-27b on 3'-UTR of sheep myostatin (MSTN) mRNA, and the double luciferase reporter assay proved that miR-27b could successfully bind on this site. When sheep satellite cells were in the proliferation status, miR-27b was upregulated and MSTN was downregulated significantly (P0.01). When miR-27b mimics was transfected into sheep satellite cells, the cell proliferation was promoted and the protein level of MSTN was significantly downregulated (P0.01). Moreover, miR-27b regulated its target gene MSTN by translation repression at an early step, and followed by inducing mRNA degradation in sheep satellite cells. Based on these results, we confirm that miR-27b could promote sheep skeletal muscle satellite cell proliferation by targeting MSTN and suppressing its expression.
机译：为了研究miR-27b在绵羊骨骼肌发育中的作用，在这里我们首先克隆了绵羊pre-miR-27b的序列，然后进一步研究了其在体内在绵羊骨骼肌中的表达模式，miR-27b表达与绵羊的关系。骨骼肌卫星细胞在体外增殖和分化，然后在此发育过程中最终确定了其靶基因。 MiR-27b序列，尤其是其成熟序列，在不同物种之间是保守的。 MiR-27b在绵羊骨骼肌中的表达高于其他组织。在萨福克和Bashbay绵羊的骨骼肌中，miR-27b在胎儿期显着上调，在产后期间显着下调（P <0.01），但仍比Bashbay保持相对高的表达水平。绵羊肌肉生长抑制素（MSTN）mRNA的3'-UTR上可能存在一个miR-27b靶位点，双荧光素酶报告基因检测证明miR-27b可以在该位点成功结合。当绵羊卫星细胞处于增殖状态时，miR-27b上调，MSTN显着下调（P <0.01）。当miR-27b模拟物转染到绵羊卫星细胞中时，细胞增殖得到促进，MSTN的蛋白水平显着下调（P <0.01）。而且，miR-27b在早期通过翻译抑制来调节其靶基因MSTN，然后在绵羊卫星细胞中诱导mRNA降解。根据这些结果，我们证实miR-27b可以通过靶向MSTN并抑制其表达来促进绵羊骨骼肌卫星细胞的增殖。

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