miR-200c对人绒毛膜滋养细胞迁移、侵袭、EMT的影响及其分子机制  

Effects of miR-200c on migration,invasion,and EMT of human chorionic trophoblast cells

作　　者：郝媛媛 徐曼 安泓润 HAO Yuanyuan;XU man;AN Hongrun(Department of Obstetrics,Hebei PetroChina Central Hospital,Langfang 065000,China)

机构地区：[1]河北中石油中心医院产科,河北廊坊065000

出　　处：《山东医药》2021年第35期31-34,共4页Shandong Medical Journal

基　　金：河北省医学科学研究课题计划项目(20201206)。

摘　　要：目的探讨miR-200c对人绒毛膜滋养细胞迁移、侵袭及上皮—间质转化(EMT)的影响,并探讨其可能的分子机制。方法将人绒毛膜滋养细胞HTR-8/SVneo分为过表达组、沉默组及对照组,应用脂质体转染法分别转染miR-200c模拟物(miR-200c mimics)、miR-200c抑制物(miR-200c inhibitor)、miRNA阴性对照(miR-NC)。采用实时荧光定量PCR法检测各组细胞miR-200c表达,细胞划痕实验和Transwell侵袭实验观察各组细胞迁移和侵袭能力(分别以细胞迁移率和侵袭细胞数表示),Westernblotting法检测各组细胞中EMT相关蛋白(E-cadherin、N-cadherin、Fibro⁃nectin)和Notch1蛋白相对表达量。双荧光素酶报告基因实验验证miR-200c与Notch1的靶向关系。结果过表达组、对照组和沉默组细胞miR-200c相对表达量依次降低,细胞迁移率和侵袭细胞数依次升高,组间两两比较P均<0.05。过表达组、对照组、沉默组E-cadherin蛋白相对表达量依次降低,N-cadherin、Fibronectin和Notch1蛋白相对表达量依次升高,组间两两比较P均<0.05。转染Notch1-WT(野生型)和miR-200c mimics的细胞相对荧光素酶活性为0.31±0.09,转染Notch1-WT和miR-NC的细胞为0.99±0.03(P<0.05);转染Notch1-MUT(突变型)和miR-200c mimics的细胞相对荧光素酶活性为1.00±0.05,转染Notch1-MUT和miR-NC的细胞为1.02±0.04(P>0.05)。结论miR-200c能够抑制人绒毛膜滋养细胞的迁移、侵袭及EMT,其机制可能与靶向调控Notch1有关。Objective To investigate the effects of miR-200c on the migration,invasion,and epithelial-mesenchy⁃mal transformation(EMT)of human chorionic trophoblast cells,and to explore the possible molecular mechanism.Meth⁃ods Human chorionic trophoblast HTR-8/SVneo cells were divided into the overexpression group,silencing group,and control group.And liposome transfection method was used to transfect miR-200c mimics,miR-200c inhibitors and miRNA negative control into cells of the above groups.The qRT-PCR was applied to detect the expression of miR-200c in each group.Cell Scratch test and Transwell invasion test were used to observe the migration and invasion abilities of cells in each group(expressed by cell migration rate and the number of invasive cells,respectively).Western blotting was used to detect the relative expression of EMT-related proteins(E-cadherin,N-cadherin,fibronectin)and Notch1 protein in cells of each group.Double luciferase reporter gene experiment was used to verify the targeting relationship between miR-200c and Notch1.Results The relative expression levels of miR-200c in the overexpression group,control group and silencing group decreased in turn,while the cell migration rate and the number of invasive cells increased in turn,with statistically significant difference between every two groups(all P<0.05).The relative expression levels of E-cadherin protein in the overexpression group,control group and silencing group decreased in turn,while the relative expression levels of N-cadherin,fibronectin and Notch1 protein increased in turn,with statistically significant difference between every two groups(all P<0.05).The relative luciferase activity of cells transfected with Notch1-WT(wild-type)and miR-200c mim⁃ics was 0.31±0.09,and that of cells transfected with Notch1-WT and miR-NC was 0.99±0.03(P<0.05).The relative lu⁃ciferase activity of cells transfected with Notch1-MUT(mutant)and miR-200c mimics was 1.00±0.05,and that of cells transfected with Notch1-MUT and miR-NC was 1. 02±0. 04(both P>0.

关 键 词：人绒毛膜滋养细胞 MIR-200C NOTCH1 细胞迁移 细胞侵袭 上皮—间质转化 

分 类 号：R714.2[医药卫生—妇产科学]