Hippo-YAP信号通路调控子宫内膜异位症上皮-间质转化的分子机制

Endometriosis is one of the common disorders that result in chronic pelvic pain and infertility in reproductive women. However, the pathogenesis is still unclear, the current treatment is not cure, and the recurrence is not low. Previous studies have revealed that epithelial-mesenchymal transition (EMT) is an essential process in pathogenesis of endometriosis, thus the endometrial epithelial cells acquire ability of cell adhere and cell migration. The underlying mechanism of EMT in endometriosis is lack of study. As a new signal pathway found recently, it has been reported that Hippo-Yes-associated protein (YAP) signal pathway play an important role in regulation of organ development and cell migration in cancer studies. Our current study found that increased expression of YAP in nucleus of endometrial epithelial cells at ectopic and eutopic endometrium of women with ovarian endometriosis. EMT transcription factor and markers have changed when knockdown or overexpress YAP in endometrial epithelial cell line in vitro. Above results indicated that Hippo-YAP signaling pathway involves in EMT of endometrial epithelial cells of endometriosis. In this current study, we plan to identify the activity of Hippo-YAP signaling pathway and EMT related transcription factors and markers in eutopic endometriosis of ovarian endometriosis; then investigate the alteration of EMT transcription factors and markers when knockdown or overexpress YAP in endometrial epithelial cell line in vitro; finally, ChIP-QPCR and Luciferase assay will be used to identify the gene loci of YAP. Our research purpose is to explicate the regulation of Hippo-YAP signal pathway on EMT of endometriosis; and further to explore a new biological target for diagnosis and treatment of endometriosis.

子宫内膜异位症(简称内异症)是引起育龄期妇女慢性盆腔痛与不孕的常见病，其发病机制不明、疗效不佳且易复发。研究表明内异症发生发展关键在于子宫内膜上皮细胞通过上皮-间质转化(EMT)实现细胞迁移、粘附和侵袭，但调控机制尚不明。近年发现的Hippo-YAP通路参与细胞迁移侵袭调控，我们发现内异症在位内膜上皮细胞核中YAP蛋白高表达，人子宫内膜上皮细胞系过表达或沉默YAP可致EMT标记物表达改变，且过表达YAP的细胞侵袭力增强，提示Hippo-YAP通路可能参与内异症EMT调控。本研究首先在人卵巢型内异症组织确定Hippo-YAP信号通路变化及其与EMT的关联，并在内膜上皮细胞沉默和过表达YAP以明确其对EMT相关因子的作用，最后通过免疫共沉淀、 ChIP-qPCR和荧光素酶实验阐明Hippo-YAP调控内膜上皮细胞EMT的分子机制，为探讨内异症发病机制及寻找新治疗靶点提供理论依据。

子宫内膜异位症(简称内异症)是引起育龄期妇女慢性盆腔痛与不孕的常见病，其发病机制不明、疗效不佳且易复发。研究表明内异症发生发展关键在于子宫内膜上皮细胞通过上皮-间质转化(EMT)实现细胞迁移、粘附和侵袭，但调控机制尚不明。近年发现的Hippo-YAP通路参与细胞迁移侵袭调控，本研究显示（一）人的在位内膜中，YAP在内异症和正常对照组的上皮和间质细胞的细胞核和细胞浆中都有分布，内异症组的在位内膜中的表达较强；E-cadherin，N-cadherin和Cytokeratin的在细胞膜和细胞浆中均有表达。（二）构建沉默YAP细胞系后，RT-qPCR和WB显示实验组YAP表达明显低于对照组，E-cadherin和Cytokeratin的表达增加；过表达YAP后，RT-qPCR显示实验组YAP表达明显增加，Cytokeratin表达增加，E-cadherin表达增加，但差异无统计学意义。（三）构建小鼠内异症模型，Verteporfin药物处理后，实验组YAP的表达量较对照组YAP表达明显降低，与YAP表达改变相一致，实验组N-cadherin的表达较对照组降低，而E-cadherin在实验组的表达较对照组升高。本研究提示Hippo -YAP通路参与子宫内膜异位症的EMT调控。