沉默Ndfip2对肝癌细胞迁移和侵袭的影响  

Effect of silencing Ndfip2 on migration and invasion of hepatocellular carcinoma cells

作　　者：张强 罗彩云 孙达权 曾志锐 郑志菊 徐国强 ZHANG Qiang;LUO Caiyun;SUN Daquan;ZENG Zhirui;ZHENG Zhiju;XU Guoqiang(School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Division of Operation,the 2 nd People's Hospital of Guiyang,Guiyang 550023,Guizhou,China;Center of Medical Research,the 3 rd Affiliated Hospital of Guizhou Medical University,Duyun 558000,Guizhou,China)

机构地区：[1]贵州医科大学基础医学院,贵州贵阳550025 [2]贵阳市第二人民医院手术室,贵州贵阳550023 [3]贵州医科大学第三附属医院医学研究中心,贵州都匀558000

出　　处：《贵州医科大学学报》2021年第8期893-899,953,共8页Journal of Guizhou Medical University

基　　金：2015年贵州省中医药、民族医药科学技术研究课题[黔中医药发(2015)8号,QZYY-2015-012];2014年度贵州省卫生计生委科学技术基金项目(gzwjkj2014-1-029)。

摘　　要：目的探究Nedd4家族交互作用蛋白2(Ndfip2)对人肝癌细胞迁移和侵袭能力的影响。方法采用免疫组织化学EnVision染色法和蛋白印迹法检测Ndfip2在肝癌组织和和肝癌细胞SMMC-7721、HepG2中的表达;在SMMC-7721和HepG2细胞中转染靶向Ndfip2小干扰RNA(si-Ndfip2)和无义siRNA序列分别构建Ndfip2基因沉默(si-Ndifip2)组和阴性对照(NC),蛋白印迹检测转染效率;采用划痕、Transwell以及形态学观察检测NC和si-Ndifip2组细胞的迁移、侵袭以及形态学变化,蛋白印迹法检测上皮-间质转化(EMT)相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)蛋白表达的影响。结果与癌旁组织比较,肝癌组织中Ndfip2表达显著上调(P<0.05);与正常肝细胞LO2细胞比较,肝癌细胞SMMC-7721和HepG2细胞中Ndfip2表达均明显上调(P<0.01);Ndfip2 si-RNA转染SMMC-7721和HepG2细胞后,Ndfip2蛋白表达均明显下调(P<0.01);划痕实验、Trasnwell实验和形态学观察结果表明,与NC组比较,si-Ndifip2组SMMC-7721和HepG2细胞迁移和侵袭能力均明显减弱(P<0.01),细胞形态上皮样转化;蛋白印迹法检测显示,与NC组比较,si-Ndifip2组SMMC-7721和HepG2细胞中N-cadherin、Vimentin蛋白表达水平均下降(P<0.05),而E-cadherin蛋白表达则增加(P<0.05)。结论沉默Ndfip2基因可以抑制肝癌细胞的迁移、侵袭能力,其机制可能与EMT过程有关。Objective To investigate the effect of Nedd4 family interacting protein 2(Ndfip2)on the migration and invasion ability of hepatocellular carcinoma(HCC)cells.Methods The expressions of Ndfip2 in hepatocellular tissues and carcinoma cells SMMC-7721,epG2 were detected by IHC EnVision and Western blot.SMMC-7721 and HepG2 cells were transfected with the specific si-Ndfip2 gene and negative siRNA sequencing to construct si-Ndfip2 and negative control(NC)cells,while the transfection efficiency was by Western blot.The migration,invasion and morphological changes of NC and si-Ndfip2 group cells were detected by wound healing assay,Transwell assay and morphology observation.The expression levels of EMT related proteins(E-cadherin,N-cadherin,and Vimentin)were detected by Western blot.Results Ndfip2 expression was obviously up-regulated in hepatic tissues and HCC cells compared with adjacent non-tumor hepatic tissues(P<0.05);compared with healthy hepatic cell LO2,the expressions of Ndfip2 protein in SMMC-7721 and HepG2 cells were obviously up-regulated(P<0.01).After Ndfip2 si-RNA transfected SMMC-7721 and HepG2 cells,expression of Ndfip2 si-RNA was significantly down-regulated(P<0.01).In wound healing test,Transwell test and morphology observation,the migration and invasion ability of SMMC-7721 and HepG2 cells in si-Ndfip2 group were significantly decreased compared with NC group(P all<0.01).Western blot showed that the protein expression levels of N-cadherin and Vimentin in si-Ndfip2 group SMMC-7721and HepG2 cells were significantly decreased compared with NC group(P all<0.05),while E-cadherin expression was significantly increased(P<0.05).Conclusion The silencing of Ndfip2 gene inhibits the migration and invasion of hepatocarcinoma cells,the mechanism may be related to the inactivation of EMT signal pathway.

关 键 词：肝肿瘤细胞 Nedd4家族交互作用蛋白2 迁移 侵袭 上皮间质转化 

分 类 号：R735.7[医药卫生—肿瘤]