Cdc42-FMNL2通过增强上皮-间充质转化促进子宫内膜异位症发生与发展的功能和机制研究

子宫内膜异位症（EMs）细胞的异位种植与上皮-间充质转化（EMT）相关，在总结前人和申请人前期工作的基础上，提出假设：异位内膜组织中高表达的Cdc42通过作用FMNL2，促进肌动蛋白的聚合和丝状伪足的伸长，上调EMT，增强异位子宫内膜细胞的侵袭和迁移，促使细胞异位种植，最终发展成为子宫内膜异位症。拟在此基础上，检测EMT标记物蛋白和RNA表达的变化；免疫荧光观察Cdc42和FMNL2敲除对于改变EMT标记物的细胞定位、肌动蛋白聚合和伪足形成的影响；免疫共沉淀技术分析Cdc42和FMNL2在原代EMs细胞和正常内膜细胞中相互结合的差异。在Cdc42敲除的细胞中过表达FMNL2，观察细胞的侵袭和迁移结果。最后，利用小鼠内异症模型，观察异位灶形成情况。上述研究结果将有助于我们进一步阐明EMs发生和发展的分子生物学机制，并为临床治疗EMs提供新的思路；同时也为补充经血逆流假说提供基础实验依据。

The ectopic implantation of endometriosis (EMs) cells is associated with epithelial-mesenchymal transition (EMT). Based on the previous work of the predecessors and the applicant, a hypothesis is proposed: high expression of Cdc42 in ectopic endometrial tissue promotes the polymerization of actin and the elongation of filopodia through FMNL2, up-regulates EMT, enhances the invasion and migration of ectopic endometrial cells promotes ectopic implantation of cells, and eventually develops to endometriosis. On this basis, the changes of EMT marker protein and RNA expression will be detected. Immunofluorescence will be used to observe the effects of Cdc42 and FMNL2 knockout on the cell localization, actin polymerization and pseudopod formation of EMT markers. Differences between the interaction of Cdc42 with FMNL2 in primary EMs cells and normal endometrial cells will be examined by CO-IP. FMNL2 will be overexpressed in Cdc42 knockout cells and cell invasion and migration results will be observed. Finally, the formation of ectopic foci is observed by using a mouse model of endometriosis. The above research results will help us further elucidate the molecular biological mechanism of EMs occurrence and development, and provide new ideas for clinical treatment of EMs. At the same time, it also provides basic experimental basis for supplementing the menstrual blood countercurrent hypothesis.