miR-409-3p靶向ZEB1负性调控卵巢浆液性癌HO-8910细胞上皮间质转化  

Mechanism of miR-409-3p negatively regulating epithelial mesenchymal transition of ovarian serous carcinoma HO-8910 cells by targeting ZEB1

作　　者：朱静 张龙辉 谢荣凯 ZHU Jing;ZHANG Longhui;XIE Rongkai(Department of Gynecology and Obstetrics,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China)

机构地区：[1]陆军军医大学(第三军医大学)第二附属医院妇产科,重庆400037

出　　处：《陆军军医大学学报》2022年第5期455-460,共6页Journal of Army Medical University

摘　　要：目的探讨miR-409-3p靶向ZEB1对卵巢浆液性癌HO-8910细胞上皮间质转化的影响及作用机制。方法利用脂质体分别将对照mimics和miR-409-3p mimics转染入HO-8910细胞中,qRT-PCR检测其转染效率,Western blot检测各组细胞中ZEB1、E-cadherin、N-cadherin和Vimentin蛋白表达水平,双荧光素酶报告基因实验检测miR-409-3p与ZEB1之间的靶向调控作用,Transwell侵袭和迁移实验分别各组细胞的侵袭和迁移能力变化。结果与mimics对照组相比,miR-409-3p过表达组中miR-409-3p表达显著升高(P<0.05),E-cadherin蛋白表达显著增加(P<0.05),ZEB1、N-cadherin和Vimentin蛋白表达明显减少(P<0.05),上皮间质转化进程被阻断。双荧光素酶报告基因实验结果显示ZEB1是miR-409-3p的直接作用靶点,过表达ZEB1可逆转转染miR-409-3p mimics介导的对细胞侵袭、迁移及上皮间质转化的抑制作用(P<0.05)。结论过表达miR-409-3p通过抑制ZEB1蛋白表达,负性调控卵巢浆液性癌HO-8910细胞上皮间质转化,抑制肿瘤细胞的侵袭和转移。Objective To investigate the effects of miR-409-3p on epithelial mesenchymal transition(EMT)of ovarian serous carcinoma HO-8910 cells by targeting ZEB1 and its underlying mechanism.Methods Control mimics and miR-409-3p mimics were transfected into HO-8910 cells by liposomes respectively,and qRT-PCR was adopted to detect the transfection efficiency.The expression of ZEB1,E-cadherin,N-cadherin,and Vimentin proteins in each cell group were examined by Western blotting,and the targeted regulation of ZEB1 by miR-409-3p was determined by dual luciferase reporter assay.Transwell invasion and migration assay was performed to observe the changes of invasion and migration abilities of the cells in each group.Results As compared with the control group,the expression of miR-409-3p was increased significantly in the miR-409-3p overexpression group(P<0.05),along with the elevation of E-cadherin expression(P<0.05),while the levels of ZEB1,N-cadherin and Vimentin were decreased remarkably(P<0.05),suggesting that the EMT process of HO-8910 cells was blocked.Furthermore,the results of dual luciferase reporter assay showed that ZEB1 was the direct target of miR-409-3p;overexpression of ZEB1 reversed the miR-409-3p mimics mediated repression of cell invasion,migration as well as EMT process(P<0.05).Conclusion Overexpression of miR-409-3p negatively regulates the EMT process by inhibiting ZEB1 expression,and thus suppresses the invasion and metastasis of ovarian serous carcinoma HO-8910 cells.

关 键 词：miR-409-3p 卵巢浆液性癌 侵袭和转移 上皮间质转化 

分 类 号：R394-33[医药卫生—医学遗传学] R73-362[医药卫生—基础医学]