热激蛋白HSP27在多囊卵巢综合征卵巢颗粒细胞增殖分化异常中的作用及分子机制

项目名称： 热激蛋白HSP27在多囊卵巢综合征卵巢颗粒细胞增殖分化异常中的作用及分子机制

项目编号： No.81501230

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： 吴亮

作者单位： 郑州大学

项目金额： 18万元

中文摘要： 热激蛋白HSP27广泛存在于人体细胞，对维持细胞内稳态至关重要。迄今对HSP27在卵巢中的功能及其与多囊卵巢综合征的关系仍知之甚少。我们的预实验结果提示HSP27可能参与卵泡刺激素FSH对卵巢颗粒细胞的增殖调控，但其作用机制还有待进一步探讨。因此，我们提出假说：HSP27可能在FSH调控颗粒细胞增殖分化中发挥重要作用，其异常将诱导多囊卵巢综合征的发生。为了验证这一假说，本研究拟以人/大鼠原代颗粒细胞和人卵巢颗粒细胞系KGN为研究模型，采用CRISPR/Cas9系统、慢病毒干扰和过表达、pull-down、体外磷酸化实验、体外甲基化/去甲基化实验和质谱等研究方法和手段，从分子、细胞和组织水平探讨HSP27在颗粒细胞增殖分化调控中的作用及分子机制。本研究将从HSP27调控颗粒细胞增殖和分化这个新视点出发，为揭示多囊卵巢综合征的发生机制奠定理论基础，为多囊卵巢综合征的临床防治提供新的思路。

中文关键词： 多囊卵巢综合征；卵泡发育；卵巢颗粒细胞；热激蛋白27；翻译后修饰

英文摘要： Heat shock protein HSP27 is ubiquitously expressed in human cells and is critically important in maintaining the cellular homeostasis. The function of HSP27 in the ovary and its association with polycystic ovary syndrome remains poorly known. Our preliminary results suggested that HSP27 might be involved in the regultion of proliferation of granulosa cells by follicle stimulating hormone (FSH), however, the functional mechanisms need to be further explored. Therefore, we hypothesize that HSP27 might have critical roles in the regulation of proliferation and differentiation of granulosa cells by FSH, and the dysfunction of HSP27 would result in the development of polycystic ovary syndrome. By utilizing the primary human/rat granulosa cells and steroidogenic human granulosa cell line KGN, combined with research methods, including CRISPR/Cas9 system, lentivirus-mediated interference and overexpression, pull-down, in vitro phosphorylation assay, in vitro methylation/demethylation assay, and mass spectrometry, this project sets out to explore the function and molecular mechanisms of HSP27 in the regulation of proliferation and differentiation of granulosa cells at molecular, cellular, and tissue levels. This study will contribute to the understanding of the pathogenesis of polycystic ovary syndrome from the novel perspective of HSP27 regulating proliferation and differentiation of granulosa cells, and provide new thoughts for clinical therapies of this disease.

英文关键词： polycystic ovary syndrome;follicular development;ovarian granulosa cells;HSP27;post-translational modification