ZEB1在卵巢子宫内膜异位症中的表达及功能研究

Expression and Function of ZEB1 in Ovarian Endometriosis

目的研究E盒增强子结合蛋白1(ZEB1)在卵巢子宫内膜异位症(ovarian endometriosis,OEMs)中的表达情况,初步探索ZEB1在子宫内膜异位症发生、发展中的作用。方法收集30例OEMs患者的卵巢囊壁组织及30例同期行手术治疗的其他妇科良性疾病患者的正常子宫内膜,分别采用实时定量PCR(qRT-PCR)及免疫组化检测ZEB1mRNA及蛋白表达水平;分别向正常子宫内膜间质细胞转染空白质粒pCI-neo(空白质粒组)及重组质粒pCI-neo-ZEB1(重组质粒组),通过CCK-8比色法测定内膜间质细胞体外增殖能力,Transwell实验检测间质细胞迁移、侵袭能力。结果ZEB1在OEMs的异位内膜间质细胞及腺体胞核中的表达较正常子宫内膜明显升高,且分泌期的表达高于增生期,差异有统计学意义(P<0.05)。应用CCK-8法及Transwell分别检测细胞增殖及侵袭能力,发现转染ZEB1表达载体后,间质细胞的增殖、侵袭能力明显增强。结论ZEB1表达上调促进异位内膜细胞的增殖能力,并增强了间质细胞的迁移、侵袭能力。

Objective To detect the expression of ZEB1 in ovarian endometriosis and evaluate its potential function.Methods The mRNA expression of ZEB1 in 30 endometriotic tissues and 30 normal endometrial was detected.The expression differences of ZEB1 protein between normal endometrium and ectopic endometrium was compared by immunohistochemical method.pCI-neo-ZEB1 eukaryotic expression vector was transfected into primary cultured endometriotic stromal cells(ESCs)and normal stromal cells by liposome.The transfection efficiency was detected by Western blot.The effects on ESCs proliferation of recombinant plasmids were detected by CCK8.Transwell assays were performed by transwell chambers to investigate the migration and invasion of ESCs.Results Up-regulated ZEB1 expressions in mRNA and protein level were observed in secretary phase of endometriotic tissues.ZEB1 expression in ectopic endometrium was obviously higher than in normal endometrium,and the difference was significant(P<0.05).After the pCI-neo-ZEB1 transfection eukaryotic expression vector,the proliferation ability of ESCs was enhanced,and the transwell migration and invasion assays showed that ZEB1 overexpression dramatically promoted ESCs migration and invasion.Conclusion Up-regulated ZEB1 in endometriotic stromal cells induced invasion and growth of endometriotic cells.