肥胖型多囊卵巢致病基因候选克隆及其致病机理的研究

The incidence of Polycystic Ovary Syndrome is about5- 6%, most of them are obesity, the genes which have been found correlating with obesity of PCOS are Ghrelin, adiponectin et.al. Whether only one or with other mutation area of Ghrelin gene contributing to the obesity of PCOS and its molecular pathogenesis need to be confirmed. In this study, relying on the large-scale PCOS genetic resource database, 500 PCOS cases are performed for mutation screening of Ghrelin by using DHPLC mutation screening platform. On the basis of the mutation 501G/C in the promoter area of Ghrelin gene, combined with new mutations to be identified in this study, we will explore the effects of the mutations of Ghrelin gene on the obesity of PCOS. We will sequence and colon the mutations gene of Ghrelin using the Miseq sequencing platform, then the plasmid carrier will be completed and which will be transfectioned to the HELA cells and then injected to the PCOS mouse model, a series of molecular pathogenesis experiments will be confirmed the mutations of Ghrelin gene correlation with the obesity of PCOS, which will provide a new theoretical basis for the genetic intervention and treatment.

多囊卵巢综合征(PCOS)在中国的发病率达5-6%,而肥胖型患者占PCOS的绝大部分,有关PCOS肥胖发病基因目前被发现的仅有Ghrelin(本研究小组发现),脂联素等,而Ghrelin是单独一个突变位点影响还是与其它突变位点连锁影响PCOS肥胖的发生,以及其影响的分子机制有待研究证实。故本研究小组依托①已经建立的大型PCOS遗传资源库，以500余例PCOS为研究对象，采用DHPLC突变/多态筛选平台对PCOS的Ghrelin基因外显子进行突变筛查，在已经发现的启动子区突变位点501G/C的基础上，结合新发现的突变位点进行连锁分析与PCOS肥胖的相关性；②采用Miseq技术平台对肥胖PCOS患者进行突变Ghrelin基因验证克隆；③构筑突变基因真核质粒表达载体，转染细胞并注射动物后进一步从分子机制验证Ghrelin突变基因参与PCOS肥胖的发生，为将来进行遗传学干预和治疗提供新的理论基础。