STAT3调控FOXL2的分子机制研究

FOXL2 is a transcription factor which is related with BPES, the development and cancer of ovary, apoptosis, stress response, cell cycle regulation and so on. As the only disease causing gene related with BPES so far, it is urgent to understand the regulatory mechanism of FOXL2. While on the basic research, it is short of exploring the transcriptional regulation mechanism, thus limiting the exhaustive understanding of FOXL2. Based on these, this project intend to: ① Taken the promoter of FOXL2 as the cut-in point, we want to find and choose the interested regulatory elements (STAT3) to research the transcriptional regulation mechanism. ② Using EMSA and ChIP to confirm the combination of STAT3 and FOXL2 in vitro and in vivo, and using QPCR, Western blot and hybridization to test the results. ③ Detecting the expression level of FOXL2 after STAT3 was inhibited using RNAi, and using Flow Cytometry to detect the states of Hela after RNAi. ④ Detecting the expression level of the downstream maker genes of the STAT3 signaling transduction pathway and FOXL2 in Hela, which was treatment with the inhibitor of STAT3 signaling transduction pathway. And combined with the luciferase reporter vector to explore the specific signaling pathway of STAT3 which regulate FOXL2. This research aim to provide new experimental basis to comprehensive understanding the nexus function of FOXL2, and further to provide theoretical foundation to use this gene in clinical.

FOXL2是一个与小睑裂综合症（BPES）、卵巢发育及癌症、细胞凋亡、抗逆响应、细胞周期等相关的转录因子，做为迄今唯一发现的BPES致病基因，在临床上迫切需要了解其调控机制，在基础研究上，现有的结果缺乏对其转录调控机制的探索，很大程度上限制了人们对其功能的全面了解。基于此，我们①以FOXL2启动子做为研究其调控机制的切入点，发现并选择感兴趣的调控元件（STAT3）深入研究；②EMSA和ChIP验证STAT3和FOXL2在体外和体内的结合，QPCR，Western blot及杂交检测结合结果；③RNAi抑制STAT3，检测FOXL2表达水平，并用流式检测细胞状态；④用STAT3信号通路抑制剂处理Hela细胞，检测下游基因及FOXL2表达水平，结合荧光素酶报告基因载体，探索调控FOXL2的信号通路。本课题将为深入了解FOXL2承上启下的功能提供实验依据，为临床上进一步利用该基因奠定理论基础。

FOXL2是一个与小睑裂综合症（BPES）、卵巢发育及癌症、细胞凋亡、抗逆响应、细胞周期等相关的转录因子，作为一个具有承上启下功能的转录因子，现有的研究侧重于探索受其调控的基因及调控机制，而缺乏对其自身转录调控机制的探索，很大程度上限制了人们对其功能的全面了解。基于此，我们①以FOXL2启动子做为研究其调控机制的切入点，发现并选择感兴趣的调控元件（STAT3）深入研究；②用EMSA和ChIP验证STAT3可以直接结合在FOXL2的启动子区域，并将结合位点精确在31 bp内；③用RNAi抑制STAT3，发现STAT3-FOXL2途径在宫颈癌细胞额增殖和凋亡方面具有重要作用，且进一步证实在细胞内存在一条Jak-STAT3-FOXL2信号通路。本课题在细胞和分子水平为完善和深入了解FOXL2承上方面的功能提供了实验依据，同时为进一步研究STAT3-FOXL2通路的功能提供了指向性结果，为临床上进一步利用该基因奠定了理论基础。