ZEB1调控DNMT1在维持小鼠表皮干细胞干性中的作用及机制研究

项目名称： ZEB1调控DNMT1在维持小鼠表皮干细胞干性中的作用及机制研究

项目编号： No.81502709

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： 汪犇

作者单位： 中南大学

项目金额： 18万元

中文摘要： DNA甲基化转移酶1(DNMT1)是一个重要的表观遗传学修饰酶，本课题组前期研究显示DNMT1可维持小鼠表皮干细胞干性，但其上游调控机制尚未报道。生物信息学预测转录因子ZEB1可与DNMT1基因启动子结合，我们的预实验亦显示下调小鼠表皮干细胞中的ZEB1基因后，DNMT1表达下降，分化相关基因K10和involucrin出现明显的上调。由此，我们推测：ZEB1可能通过与DNMT1启动子结合，调控干细胞分化相关基因的甲基化水平，影响小鼠表皮干细胞的干性。本研究中，我们将以小鼠表皮干细胞为研究对象，首先利用慢病毒载体技术调节ZEB1的表达量，检测ZEB1对小鼠表皮干细胞干性指标的影响；其次研究ZEB1调控DNMT1对小鼠表皮干细胞干性的作用及可能机制，并进一步确定ZEB1结合DNMT1的区域；最后，利用ZEB1局部过表达或敲除技术在体内验证上述机制，为探索表皮干细胞治疗研究领域提供新的方向。

中文关键词： 表皮干细胞；ZEB1；DNA甲基化转移酶1；干性；DNA甲基化

英文摘要： DNA methyltransferase 1 (DNMT1) is an important epigenetic modifying enzyme. Our previous research has shown DNMT1 could maintain the stemness of mouse epidermal stem cells, but the regulatory mechanism upstream has not yet been reported. According to the prediction database, transcription factor ZEB1 can combine with the promoter region of DNMT1. We found in our pre-experimet that knockdown the expression of ZEB1 in the mouse stem cells can decrease DNMT1 expression and increase K10 and involucrin expression. We infer that DNMT1 play an important part in maintaining the stemness of mouse epidermal stem cells regulated by binding to ZEB1. In the present study, we will clarify the role of ZEB1 in regulating mouse stem cells stemness by regulating expression of ZEB1. And how ZEB1 affects on stemness of mouse stem cells through binding to DNMT1 will be elaborated thoroughly in the aspects of transcription factor-DNA promoter interaction. Finally, the mechanism will be verified in ZEB1 overexpression or downexpression mice in vivo. Our study aims to explore the roles and mechanism of DNMT1 regulation by ZEB1 in maintaining the stemness of mouse epidermal stem cells, then provide a new direction in the research field and therapeutic field of epidermal stem cells.

英文关键词： epidermal stem cells;ZEB1;DNMT1;stemness;DNA methylation