摘要
目的探究miR-576-5p调控上皮细胞-间充质转化(epithelial-mesenchymal transition,EMT)进程在肝癌细胞迁移和侵袭中的影响。方法采用RT-qPCR检测重庆医科大学附属第一医院肝胆外科2020年6月至2021年1月行肝癌切除术的27例患者的肝细胞癌组织(hepatocellular carcinoma,HCC)和癌旁组织,同时检测了肝癌细胞株和正常人肝细胞的miR-576-5p表达水平。将miR-576-5p类似物/阴性对照(miR-576-5p mimics/mimics NC)转染到hepG2作为过表达组(mimics组)和过表达对照组(mimics NC组);将miR-576-5p抑制剂/阴性对照(miR-576-5p inhibitor/inhibitor NC)转染至SMMC-7721中作为去表达组(inhibitor组)及去表达对照组(inhibitor NC组)。用CCK-8和EDU检测细胞增殖能力;划痕实验、Transwell侵袭实验测定各组细胞侵袭和迁移能力;Western blot检测各组细胞中E-cadherin、N-cadherin和snail蛋白表达水平变化。结果miR-576-5p在肝细胞癌组织中较配对癌旁组织表达明显下降(P<0.001),在肝癌细胞株中较正常人肝细胞表达偏低(P<0.001)。SMMC-7721 inhibitor组较inhibitor NC组增殖能力增强(P<0.001,P<0.001),伤口愈合率显著增强(P<0.001),transwell小室迁移和侵袭穿孔细胞数明显增加(P<0.001,P<0.01)。mimics组较过表达对照组增殖能力减弱(P<0.01,P<0.01),伤口愈合率显著降低(P<0.01),迁移和侵袭穿孔细胞数明显减少(P<0.001,P<0.01)。miR-576-5p过表达组与其对照组比较,E-cadherin蛋白表达显著增加,N-cadherin和snail蛋白表达明显减少(P<0.001,P<0.05,P<0.05),miR-576-5p去表达则发生相反的变化(P<0.05,P<0.001,P<0.01)。结论miR-576-5p抑制肝癌细胞的增殖,同时阻碍EMT进程,抑制肝癌细胞侵袭和迁移。
Objective To investigate the effect of miR-576-5P on the migration and invasion of hepatocellular carcinoma(HCC)cells by regulating epithelial-mesenchymal transition(EMT)process.Methods RT-qPCR was used to detect miR-576-5p expression level in HCC cells and normal liver cells,hepatocellular carcinoma tissues and paraneoplastic tissues from 27 patients who accepted hepatectomy from June 2020 to January 2021 in the Department of Hepatobiliary Surgery of the First Affiliated Hospital of Chongqing Medical University.MiR-576-5p mimics and mimics NC were transfected into hepG2 cells as the over-expression group(mimics group)and the control group(mimics NC group),and miR-576-5p inhibitor and inhibitor NC were transfected into SMMC-7721 cells as the de-expression group(inhibitor group)and the control group(inhibitor NC group).CCK-8 assay and EDU assay were used to detect the proliferative ability of cells.Transwell assay and wound healing assay were used to observe the changes of invasion and migration of cells in each group.The changes of expression level of E-cadherin,N-cadherin and snail in each group were detected by Western blotting.Results The expression of miR-576-5p in HCC tissues was significantly lower than that in matched adjacent tissues(P<0.001),and the expression of miR-576-5p was lower in HCC cells than that of normal liver cells(P<0.001).SMMC-7721 inhibitor group had enhanced proliferative capacity(P<0.001,P<0.001),significantly increased wound healing rate(P<0.001)and significantly increased migration and invasive perforated cells(P<0.001,P<0.01),compared to the inhibited NC group.HepG2 mimics group had lower proliferative capacity(P<0.01,P<0.01),significantly lower wound healing rate(P<0.01),and significantly fewer migration and invasive perforated cells than the mimics NC group(P<0.001,P<0.01).The hepG2 mimics group showed a obviously increase in the expression of E-cadherin and a decrease in the expression of N-cadherin and snail compared to the mimics NC group(P<0.001,P<0.05,P<0.05),while the complet
作者
崔琦
王余彦
卢映君
曾爱中
CUI Qi;WANG Yuyan;LU Yingjun;ZENG Aizhong(Department of Infectious Disease,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2023年第4期366-372,F0003,共8页
Journal of Army Medical University
关键词
miR-576-5p
肝癌
上皮间质转化
miR-576-5p
hepatocellular carcinoma
epithelial mesenchymal transition
