MiRNA-224参的-研究小鼠卵泡发育调控作用的-研究.pdf

摘 要 综合以上研究结果，我们认为miR-224正是通过靶向于不同的靶基因实现 TGF- β1对卵泡发育调节的介导作用。而对miRNA参与卵泡发育过程的进一步研究 将会为人工干预生殖过程提供研究基础，如治疗生殖紊乱等。 关键词： microRNA-224 小鼠 卵泡发育 细胞增殖 COC 扩展 II Abstract ABSTRACT Many members of the TGF-β superfamily are indicated to play important roles in mammalian reproduction, such as affecting the early folliculogenesis, granulosa cell proliferation and differentiation, oocyte maturation and embryo implantation and the maintanence of pregnancy. The perturbation of the TGF-β signaling transduction could result in female infertility. MicroRNAs (miRNAs), as small non-coding RNAs, were recently found to regulate gene expression at the post-transcriptional levels, either inhibit or promote. However, little is known about the role of miRNAs in TGF-β-mediated granulosa cell proliferation，hormone release and COC expansion. In this study, the miRNA expression profiling was identified from TGF-β1-stimulated mouse preantral granulosa cells (GCs), and 3 miRNAs were found to be significantly up-regulated and 13 miRNAs were significantly down-regulated. Among the up-regulated miRNAs, miR-224 was the second most significantly elevated miRNA. This up-regulation was attenuated by treatment of GCs with SB431542 (an inhibitor of TGFβ superfamily type I receptors, thus blocking the phosphorylation of the downstream effectors Smad2/3), indicating that miR-224 expression was regulated by the canonical TGF-β1/Smads pathway. By a series studies of luciferase analysis and western blot, we idendified that smad4 is one of the target of miR-224. The ectopic expression of miR-224 by transfecting with miR-224 mimics can enhance TGF-β1-induced GC proliferation through targeting smad4. Inhibition of endogenous miR-224 partially suppressed GC proliferation induced by TGF-β1. In addition, both miR-224 and TGF-