Shi Wei, Wang Xiaoling, Ruan Lihong, Fu Jiamei, Liu Fang, Qu Jinfeng
Pharmazie. 2017 Nov 1;72(11):694-699. doi: 10.1691/ph.2017.7649.
Endometrial cancer is the most common gynecological cancer. Epithelial-mesenchymal transition (EMT) plays a critical role in tumor invasion and metastasis, which limits the success of treatment. Here, we investigated the roles of forkhead box A2 (FOXA2) and microRNA-200a (miR-200a) in regulating the EMT of endometrial cancer cells RL95-2. Empty vector or FOXA2 was stably transfected into RL95-2 cells. MTT assay measured cell proliferation, apoptosis assay measured apoptosis, Transwell invasion assay measured cell invasion, and Western blot measured the protein expression of FOXA2, E-cadherin, and vimentin. ChIP assay determined the binding of FOXA2 to E-cadherin promoter. For miR-200a analysis, the cells with stable FOXA2 expression were transfected with miR-negative control or miR-200a. Forced expression of FOXA2 decreased the proliferation and invasion, and increased the apoptosis of RL95-2 cells. FOXA2 also affected the EMT-associated proteins: FOXA2 increased the protein expression of E-cadherin and decreased the expression of vimentin. Moreover, FOXA2 positively regulated the promoter of E-cadherin in RL95-2 cells. Luciferase reporter assay identified FOXA2 as a target of miR-200a, which negatively regulated FOXA2. Western blot results showed that overexpression of miR-200a decreased the expression of E-cadherin but increased the expression of vimentin in the endometrial cancer cells by downregulating FOXA2 expression. FOXA2 may act as a tumor suppressor and inhibit EMT of endometrial cancer cells. FOXA2 expression is controlled by miR-200a, which promotes EMT of the endometrial cancer cells.
子宫内膜癌是最常见的妇科癌症。上皮-间质转化（EMT）在肿瘤侵袭和转移中起关键作用，这限制了治疗的成功率。在此，我们研究了叉头框A2（FOXA2）和微小RNA-200a（miR-200a）在调节子宫内膜癌细胞RL95-2的EMT中的作用。将空载体或FOXA2稳定转染至RL95-2细胞中。MTT法检测细胞增殖，凋亡检测法检测凋亡，Transwell侵袭试验检测细胞侵袭，蛋白质印迹法检测FOXA2、E-钙黏蛋白和波形蛋白的蛋白表达。染色质免疫沉淀（ChIP）试验确定FOXA2与E-钙黏蛋白启动子的结合。对于miR-200a分析，将稳定表达FOXA2的细胞用miR阴性对照或miR-200a转染。FOXA2的强制表达降低了RL95-2细胞的增殖和侵袭，并增加了其凋亡。FOXA2还影响与EMT相关的蛋白：FOXA2增加了E-钙黏蛋白的蛋白表达并降低了波形蛋白的表达。此外，FOXA2在RL95-2细胞中正向调节E-钙黏蛋白的启动子。荧光素酶报告基因试验确定FOXA2是miR-200a的靶标，miR-200a对FOXA2起负调节作用。蛋白质印迹结果显示，miR-200a的过表达通过下调FOXA2表达降低了子宫内膜癌细胞中E-钙黏蛋白的表达，但增加了波形蛋白的表达。FOXA2可能作为一种肿瘤抑制因子并抑制子宫内膜癌细胞的EMT。FOXA2的表达受miR-200a控制，miR-200a促进子宫内膜癌细胞的EMT。
