H19/miR-194/IGF1R/E2F1正反馈回路介导多囊卵巢综合征颗粒细胞凋亡的分子机制研究

Polycystic ovary syndrome（PCOS）is one of the most important causes of female infertility. It is also the most common endocrine disease in women of childbearing age, characterized by hyperandrogenism, polycystic ovary, persistent anovulation or rare ovulation. In this study, we analyzed the lncRNA spectrum of differentially expressed ovarian granulosa cells from patients with PCOS and normal samples, and selected lncRNA H19 with high abundance and high variation. Through bioinformatics analysis and preliminary experiments, we found that H19 can participate in the number and function maintenance of ovarian granulosa cells. It is speculated that, H19 can be used as ceRNA competitive combined with miR-194, lifting the inhibitory effect of miR-194 on IGF1R, affecting follicular development. At the same time, IGF1R can also promote the expression of E2F1, a transcription factor of H19 promoter region, as a positive feedback factor of H19. This .project is the first attempt to explore the biological function of H19 and PCOS related molecular mechanisms, in order to apply H19 clinic, and provide a theoretical basis for the diagnosis and potential therapeutic targets of PCOS in women.

多囊卵巢综合征是造成女性不孕的重要原因之一，也是育龄期女性最常见的内分泌疾病，表现为高雄激素血症、多囊卵巢、持续无排卵或稀发排卵。本课题组从分析多囊卵巢综合征患者和正常样本来源的卵巢颗粒细胞中差异表达的lncRNA谱，从中挑选出了高丰度、高变化的lncRNA H19。通过生物信息学分析和前期预实验，我们发现H19能够参与卵巢颗粒细胞的数量和功能维持。并推测，H19能够作为ceRNA竞争性结合miR-194，解除miR-194对IGF1R的抑制作用，影响卵泡的发育。同时，IGF1R还能促进H19启动子区转录因子E2F1的表达，作为H19的正反馈因子。本课题首次尝试探讨H19与多囊卵巢综合征相关的生物学功能及分子机制，以期将H19应用于临床，为女性多囊卵巢综合征的诊断和潜在治疗靶点的筛选提供理论依据。

多囊卵巢综合征(polycystic ovarian syndrome，PCOS)是造成女性不孕的重要原因之一。卵巢颗粒细胞的数量和功能是保证卵泡发育的必要条件，其增殖抑制或过度凋亡是导致卵泡发育异常和不孕症发病的重要原因。在前期研究中发现多囊卵巢综合征患者中 lncRNA H19 的水平存在差异性表达，但其与颗粒细胞的增殖凋亡关系尚不明确。本研究通过生物信息学分析筛选出目的靶基因，通过细胞转染、TUNEL、流式细胞技术检测细胞凋亡等实验，结果表明H19在PCOS患者颗粒细胞中的表达水平显著下降；在人卵巢颗粒细胞KGN中，低表达H19后能够明显的抑制KGN细胞的活性，过表达H19能够调控KGN细胞中增殖相关的蛋白，进而参与KGN细胞增殖；并且揭示H19通过miR-194/HB-EGF回路在调控卵巢颗粒细胞功能并参与多囊卵巢综合症发病的分子机制。本课题首次尝试探讨H19与多囊卵巢综合征相关的生物学功能及分子机制，为女性多囊卵巢综合征的诊断和潜在治疗靶点的筛选提供理论依据。