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.
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)
出处
《贵州医科大学学报》
CAS
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)。
