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首页> 外文期刊>BMC Biotechnology >AAV2-mediated follistatin overexpression induces ovine primary myoblasts proliferation
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AAV2-mediated follistatin overexpression induces ovine primary myoblasts proliferation

机译：AAV2介导的卵泡抑素过表达诱导绵羊原代成肌细胞增殖

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Background Follistatin (FST) has been shown to bind to some TGF-β family members and can function as a potent myostatin (MSTN) antagonist. Recent studies have revealed that over-expression of FST by adeno-associated viruses increases muscle growth in mice, humans and nonhuman primates. In the present study, to determine the effect of FST on ovine primary myoblast (OPM) proliferation, FST was over-expressed using an adeno-associated virus serotype 2 (AAV 2) vector. Results Western blot results showed that AAV induced the expression of FST protein in transduced OPM cells. Real-time quantitative PCR results indicated that over-expression of FST resulted in a dramatic increase in Akt I and CDK2 expression and a decrease in p21 expression. Moreover, cell cycle analysis confirmed that FST down-regulated p21, a CDK inhibitor, and increased the level of CDK2 expression in OPM cells. Hence, follistatin positively regulated the G1 to S progression. Our results showed that FST induced proliferation through a down-regulation of p21, as only the p21 expression level was down-regulated as a result of FST over-expression in myoblasts, whereas no change was observed in the level of p57 expression. Conclusions These results expanded our understanding of the regulation mechanism of FST in ovine primary myoblasts. Our results provide the first evidence that the AAV viral system can be used for gene transfer in ovine myoblast cells. Moreover, the results showed that an AAV vector can successfully induce the expression of FST in OPM cells in vitro. These findings demonstrated that FST over-expression induces proliferation through a down-regulation of the p21 gene under proliferating conditions.
机译：背景卵泡抑素（FST）已显示与某些TGF-β家族成员结合，可作为强效肌生长抑制素（MSTN）拮抗剂发挥作用。最近的研究表明，腺相关病毒过度表达FST会增加小鼠，人类和非人类灵长类动物的肌肉生长。在本研究中，为了确定FST对绵羊原代成肌细胞（OPM）增殖的影响，使用腺相关病毒血清型2（AAV 2）载体过表达FST。结果Western blot结果表明，AAV诱导了OPT细胞中FST蛋白的表达。实时定量PCR结果表明，FST的过度表达导致Akt I和CDK2表达急剧增加，而p21表达减少。此外，细胞周期分析证实FST会下调p21（一种CDK抑制剂）并增加OPM细胞中CDK2表达的水平。因此，卵泡抑素可正向调节G1至S的进程。我们的结果表明FST通过下调p21诱导增殖，因为成肌细胞中FST过表达仅下调了p21表达水平，而未观察到p57表达水平的变化。结论这些结果扩大了我们对绵羊原代成肌细胞中FST调控机制的认识。我们的结果提供了第一个证据，表明AAV病毒系统可用于绵羊成肌细胞中的基因转移。而且，结果表明，AAV载体可以在体外成功诱导OPM细胞中FST的表达。这些发现证明FST的过表达通过在增殖条件下下调p21基因来诱导增殖。

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