HuR/ELAVL1在大鼠卵巢颗粒细胞增殖调控中的作用及机制研究

项目名称： HuR/ELAVL1在大鼠卵巢颗粒细胞增殖调控中的作用及机制研究

项目编号： No.31500943

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 韩莹莹

作者单位： 北京林业大学

项目金额： 20万元

中文摘要： RNA结合蛋白ELAVL1，又名HuR，通过调控细胞周期相关基因的转录后表达参与多种细胞增殖过程，是卵巢癌等肿瘤疾病的潜在标志物，然而关于HuR在卵巢颗粒细胞增殖调控中的研究，迄今国内外尚无报道。我们前期研究证实，大鼠卵巢颗粒细胞中HuR蛋白的表达及核质分布与有丝分裂进程密切相关，并且受FSH和PKA信号影响，而FSH激活PKA信号对颗粒细胞增殖有重要的促进作用。据此我们推测：HuR蛋白可能通过介导细胞周期相关基因的转录后表达参与卵巢颗粒细胞增殖过程。为证实该假说，本项目拟以大鼠为研究对象，首先应用过表达和干扰技术，探究HuR蛋白对卵巢颗粒细胞增殖的影响；其次利用RIP及定点突变技术，研究PKA介导HuR磷酸化修饰对细胞周期相关基因转录后表达的影响，进而揭示HuR影响卵巢颗粒细胞增殖的分子机制。研究结果将进一步阐明哺乳动物卵巢颗粒细胞增殖及卵泡发育机制，为寻找卵巢疾病的药物靶点提供新思路。

中文关键词： 颗粒细胞增殖；HuR/ELAVL1；细胞周期调控；蛋白激酶A；转录后调控

英文摘要： RNA-binding protein ELAVL1, also named HuR, is involved in the control of various cell proliferation processes by post-transcriptional regulation of cell cycle related genes expressions. HuR is a candidate biomarker for the diagnose of ovarian cancer and other carcinomas. However, the effects and mechanisms of HuR on ovarian granulosa cell proliferation haven’t been reported. Recent studies indicated that the expression and histological localization of HuR protein were related to the phases of cell cycle during rat ovarian granulosa cells proliferation, and these phenomena were influenced by both FSH and PKA. In our previous studies, it has been demonstrated that PKA was involved in FSH-stimulated rat ovarian granulosa cell proliferation. Based on those findings, a hypothesis has been proposed that the post-transcriptional effects of HuR on the expression of cell cycle related genes may be involved in the regulation of ovarian granulosa cell proliferation. Thus, rat models will be taken to demonstrated the hypothesis. Firstly, overexpression and RNA interference experiments will be utilized to investigate the effects of HuR on the processes of rat ovarian granulosa cell proliferation. Secondly, to address the mechanisms underlying HuR-mediated ovarian granulosa cell proliferation, the effects of PKA on the phosphorylation of HuR and post-transcriptional regulation of cell cycle related genes expressions in granulosa cell will be studied by RIP and Gene-directed mutagenesis. These findings will advance the current understanding on the regulation mechanism of ovarian granulosa cell proliferation and follicle development, and also provide new insights into finding a new drug target for ovarian diseases.

英文关键词： Granulosa cell proliferation;HuR/ELAVL1;Cell cycle regulation;PKA;post-transcriptional regulation