长链非编码RNA在人卵巢早衰发生中的作用机理及分子机制研究

项目名称： 长链非编码RNA在人卵巢早衰发生中的作用机理及分子机制研究

项目编号： No.81501228

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 姚桂东

作者单位： 郑州大学

项目金额： 18万元

中文摘要： 卵巢早衰严重影响女性生育和生活质量，目前对其发生的分子机制知之甚少，尤其是长链非编码RNA（lncRNA）调控卵巢早衰发生的研究尚未见报道。我们前期发现早衰卵巢组织lncRNAs表达谱异常，且这些lncRNAs多高表达于女性生殖系统中，调控颗粒细胞增殖的相关因素可影响此类lncRNAs的表达，提示lncRNAs可能参与卵巢早衰的发生。为此，我们提出假说：lncRNA 可能通过影响颗粒细胞增殖等作用导致卵巢早衰。为验证此假说，我们将通过人颗粒细胞系KGN、原代颗粒细胞和卵巢组织，采用生物信息学分析、RNA干扰、流式细胞术、RNA荧光原位杂交、RNA免疫共沉淀等手段，从分子、细胞及组织水平等多方面探讨lncRNA在卵巢早衰发生中的重要作用及其分子调控机制。本研究将从lncRNA这个新视角为揭示卵巢早衰发病机制奠定基础，并为全新的基于lncRNA的诊断和治疗女性卵巢功能衰退及并发症提供新的思路。

中文关键词： 卵巢早衰；长链非编码RNA；颗粒细胞；卵巢功能紊乱

英文摘要： Premature ovarian failure (POF) is a common female disease which severely affect female fertility and life quality, however, the reason for the cause of such disease now is poorly understood, especially the lack of long non-coding RNA (lncRNA) in the regulation of human POF. We previously found that the expression profile of lncRNAs between ovarian tissues from patients with POF and normal fertility were significantly different, and these differentially expressed lncRNAs were mostly highly expressed in female reproductive systems, such as oviduct tissues, granulosa cells and ovarian tissues. In addition, some associated factors related to the regulation of granulosa cell proliferation was also affect the expression of some of these differentially expressed lncRNAs. Therefore, these results may indicate that these differentialy expressed lncRNAs be involved in the occurrence of human POF, but the specific mechanisms are still not understood. Thus, we propose the hypothesis that the lncRNA may regulate the biological function of granulosa cells, such as proliferation, then leading to the occurrence of POF. In order to validate this hypothesis, materials of human granulosa cell line KGN, primary granulosa cell and ovarian tissue were used to investigate the important role of lncRNA in the regulation of human POF and its molecular mechanisms by using bioinformatical analysis, real-time PCR, western blot, RNA interference, flow cytometry, RNA fluorescence in situ hybridyzation and RNA immunoprecipitation, through levels of molecular, cellular and histological. This study will provide a new vision for the foundations of uncovering the important role lncRNA in the occurrence mechanism of POF, and provide a new way to diagnosis and treatment of female POF and its complications based upon the aspects of lncRNA.

英文关键词： Premature ovarian failure;Long non-coding RNA;Granulosa cell;Ovarian dysfunction