过表达miR-198通过靶向调控Wnt2B表达抑制滋养细胞HTR-8/SVneo的增殖、转移与侵袭  

Overexpression of miR-198 inhibits the proliferation,migration and invasion of trophoblast HTR-8/SVneo cells by targeting Wnt2B

作　　者：钟黎黎[1] 盛莹[1] 郭江虹[1] 阳双健 Zhong Lili;Sheng Ying;Guo Jianghong(Department of Obstetrics,the First Affiliated Hospital of Nanhua University,Hunan 421001)

机构地区：[1]南华大学附属第一医院产科,衡阳421001

出　　处：《现代妇产科进展》2021年第2期100-105,共6页Progress in Obstetrics and Gynecology

摘　　要：目的研究过表达miR-198对人滋养细胞株HTR-8/SVneo增殖、转移和侵袭能力的影响,并探讨其可能作用机制。方法qRT-PCR检测子痫前期(PE)患者及正常产妇胎盘组织中miR-198表达水平。培养人滋养细胞株HTR-8/SVneo,采用脂质体法将miR-198 mimics及其阴性对照mimics NC转染至HTR-8/SVneo细胞中,qRT-PCR检测细胞中miR-198、Wnt2B mRNA表达水平,CCK-8法检测细胞增殖活性,Transwell实验检测细胞迁移与侵袭能力,Western blot检测细胞中Wnt2B、E-cadherin、N-cadherin和Vimentin蛋白表达,双荧光素酶报告基因验证miR-198与Wnt2B的靶向关系。再将miR-198 mimics和Wnt2B过表达质粒分别或同时转染至HTR-8/SVneo细胞中,Western blot检测细胞中Wnt2B蛋白表达变化,CCK-8法和Transwell实验分别检测细胞增殖活性以及细胞迁移与侵袭能力。结果与正常组比较,PE患者胎盘组织中miR-198表达水平显著升高(P<0.001)。miR-198过表达可显著抑制HTR-8/SVneo细胞增殖活性以及细胞迁移与侵袭能力(P<0.01,P<0.001),促进细胞中E-cadherin蛋白表达(P<0.001),抑制细胞中Wnt2B、N-cadherin和Vimentin等蛋白表达(P<0.001)。双荧光素酶报告基因实验证实,miR-198靶向负调控Wnt2B基因表达。过表达Wnt2B可显著逆转miR-198对HTR-8/SVneo细胞增殖以及迁移与侵袭能力的抑制作用(P<0.001)。结论过表达miR-198可抑制人滋养细胞HTR-8/SVneo的增殖、转移与侵袭,其机制可能与靶向抑制Wnt2B基因表达有关。Objective:To investigate the effect and mechanism of overexpression of miR-198 on the proliferation,migration and invasion of human trophoblast line HTR-8/SVneo.Methods:The expression of miR-198 in placenta of preeclampsia(PE)patients and normal pregnant women was detected by qRT-PCR.Human trophoblast HTR-8/SVneo cell line was cultured.MiR-198 mimics and its negative control mimics NC were transfected into HTR-8/SVneo cells by liposome method.The expressions of miR-198 and Wnt2B mRNA were detected by qRT-PCR,cell proliferation was detected by CCK-8,cell migration and invasion were detected by Transwell,and Wnt2B,E-cadherin,N-cadherin and vimentin were detected by Western blot.Double luciferase reporter gene system analysis was used to verify the targeting relationship between miR-198 and Wnt2B.Then miR-198 mimics and Wnt2B overexpression plasmids were transfected into HTR-8/SVneo cells respectively or simultaneously.Western blot was used to detect Wnt2B protein expression.CCK-8 and Transwell were used to detect cell proliferation,cell migration and invasion.Results:Compared with the normal group,the expression of miR-198 in placenta of PE patients was significantly increased(P<0.001).Overexpression of miR-198 significantly inhibited the proliferation,migration and invasion of HTR-8/SVneo cells(P<0.01,P<0.001),promoted the protein expression of E-cadherin(P<0.001),and inhibited the expression of Wnt2B,N-cadherin and vimentin in cell(P<0.001).Double luciferase reporter gene experiment confirmed that miR-198 targeted negative regulation of Wnt2B expression.However,overexpression of Wnt2B significantly reversed the inhibitory effect of miR-198 on the proliferation,migration and invasion of HTR-8/SVneo cells(P<0.001).Conclusion:Overexpression of miR-198 can inhibit the proliferation,migration and invasion of HTR-8/SVneo cells,which may be relate to the targeted inhibition of Wnt2B expression.

关 键 词：miR-198 Wnt2B 滋养细胞 增殖 转移 侵袭 

分 类 号：R714.2[医药卫生—妇产科学]