Wnt/β-catenin信号通路在人类胚胎干细胞向子宫内膜样细胞分化过程中的调控作用及机制研究

Endometrium is a highly renewable organization. Many diseases of endometrium are related to some gynecology diseases, which cause endomentrial injure and the difficulty in endomentrial repair. However the mechanism of endometrial repair and regeneration is unclear so far. Present study confirmed that the Wnt signaling pathwayplayed an important role in regulating the development of female reproductive system. By using the hES cell line，we want to demonstrate the expression and the role of canonical Wnt pathway in endometrium differentiation of hES cclls. We intend to identify the morphological characteristics and function of the differentiated cells through immunofluorescence, RT-PCR and Western-blot and detect the expression of genes related to Wnt/β-catenin signaling pathway. Then being used activators,inhibitors,lentivirus infect and siRNA transfection to intervene the canonical Wnt signaling pathway, the differention of endometrial-like cells from embryonic stem cell was examined. This project will help to explore the role of Wnt signaling pathways in endometrial repair and the pathogenesis of abnormal endometrial proliferation diseases, and to lay the foundation of exploring therapeutic targets for diseases associated endometrium of Wnt pathways.

子宫内膜是一高度可再生组织,子宫内膜一些疾病是由于子宫内膜损伤和修复重建过程异常引起,但子宫内膜修复再生的机制迄今还不甚清楚。经典Wnt通路在干细胞的增殖、分化、迁移等方面起着重要的作用。我们在前期工作和预实验的基础上，将胚胎干细胞在子宫内膜条件培养液的体外诱导体系中培养，使胚胎干细胞向子宫内膜细胞分化，运用细胞免疫荧光法、RT-PCR及Western-blot对分化细胞进行形态及功能等方面的鉴定，同时检测Wnt/β-catenin信号通路上的相关基因的表达，并运用Wnt激活剂/抑制剂、慢病毒感染、siRNA靶向沉默等干预诱导体系中经典Wnt通路活化水平，观察人胚胎干细胞向子宫内膜细胞分化的变化，以探讨经典Wnt通路在此过程中的调控作用及机制，有助于进一步探讨Wnt通路在子宫内膜修复中的作用以及子宫内膜增殖异常相关疾病的病理机制，也为探讨Wnt通路作为子宫内膜相关疾病的治疗靶点奠定基础。

子宫内膜是一高度可再生组织,子宫内膜一些疾病是由于子宫内膜损伤和修复重建过程异常引起,但子宫内膜修复再生的机制迄今还不甚清楚。经典Wnt通路在干细胞的增殖、分化、迁移等方面起着重要的作用。我们在前期工作和预实验的基础上，将胚胎干细胞在子宫内膜条件培养液的体外诱导体系中培养，使胚胎干细胞向子宫内膜细胞分化，运用细胞免疫荧光法、RT-PCR及Western-blot对分化细胞进行形态及功能等方面的鉴定，同时检测Wnt/β-catenin信号通路上的相关基因的表达，并运用Wnt激活剂/抑制剂、慢病毒感染、siRNA靶向沉默等干预诱导体系中经典Wnt通路活化水平，观察人胚胎干细胞向子宫内膜细胞分化的变化，以探讨经典Wnt通路在此过程中的调控作用及机制；而且，通过鼠体内实验观察到干细胞分化为子宫内膜细胞，并修复受损子宫内膜恢复其妊娠功能。此研究，有助于进一步探讨Wnt通路在子宫内膜修复中的作用以及子宫内膜增殖异常相关疾病的病理机制，也为探讨Wnt通路作为子宫内膜相关疾病的治疗靶点奠定基础。