Prader-Willi位点的lncRNA对 PCOS患者卵子发育的调控研究

The aim of the project is to investigate the roles of Prader-Willi reginon non-protein coding RNA 2 (PWRN2) in controlling the oocyte development of PCOS patients. PCOS is a genetic heterogeneity disease. In our previous research, 2830 differently expressed lncRNAs were isolated from cumulus cells of PCOS patients by using cDNA microarray. Of them, one lincRNA is Prader-Willi region non-protein coding RNA 2 (PWRN2). PWRN2 was proved to be related with oocyte maturation and embryo development in PCOS patients. But the molecular mechanism is not clear. In this study, the expression of PWRN2 will be knocked down by RNA interference technology in primary cultured cumulus cells of PCOS patients and then the target genes of PWRN2 will be isolated by cDNA microarray. The interaction between PWRN2 and the target genes during oocyte development in PCOS patients will be elucidated by various molecular biology methods. Based on this research, the understanding of molecular mechanisms governing the process of oocyte development in PCOS patients might be improved.

本研究旨在探讨Prader-Willi位点的lncRNA（PWRN2）在调控PCOS患者卵子发育中的分子作用。PCOS是由多基因调控的遗传异质性疾病，我们前期采用基因芯片技术分离到与PCOS相关的lncRNAs 2830个，其中PWRN2为处于Prader-Willi位点的lincRNA，且证明PWRN2与PCOS患者的卵子成熟及胚胎发育潜能相关，但其作用机制尚不清楚。本研究将采用RNAi技术在PCOS患者的原代颗粒细胞中干扰PWRN2的表达，通过基因芯片技术筛选可能与PWRN2相互作用的与卵子发育相关的差异基因；通过多种分子生物学手段研究PWRN2对靶基因的调控机理，初步揭示Prader-Willi位点的lncRNA对PCOS患者卵子发育的分子作用，该项目的研究结果将为揭示PCOS患者卵子发育异常的机理提供新的分子依据。

多囊卵巢综合征（polycystic ovary syndrome, PCOS）是一种复杂的、异质性内分泌紊乱综合征，卵泡发育异常一直被认为是PCOS最基本的特征。课题组前期研究结果发现，PWRN2在PCOS卵丘颗粒细胞中显著高表达，推测可能与PCOS的卵子发育相关。该项目通过在KGN细胞系中干扰PWRN2的表达，采用基因芯片技术筛选到与PWRN2相关的差异mRNA，结合前期PCOS卵丘颗粒细胞miRNA和mRNA的芯片结果，确定了PWRN2-miR-92b-TMEM120B间的ceRNA作用机制，初步揭示了PWRN2在调控卵子发育过程中的作用。此外，该项目还进行了miR-509-3p对PCOS颗粒细胞中雌激素合成的调控作用研究，及miR-135b通过靶向LAST2，进而影响Hippo信号通路的正常作用，从而导致PCOS的发生的相关研究。该项目的研究结果对于深入揭示PCOS的发生机制及实现对PCOS精准治疗具有重要的理论价值。