miR-203a-3p通过SLUG调控上皮间质化抑制肝癌细胞的迁移及侵袭  

miR-203a-3p inhibits migration and invasion of HCC cells by regulating epithelial mesenchymal transition via SLUG

作　　者：张国柄[1] 徐江海[1] 王东风[1] ZHANG Guo-bing;XU Jiang-hai;WANG Dong-feng(Department of Hepatology Division 3rd,the Fifth People's Hospital of Anyang,Henan 455000,China)

机构地区：[1]安阳市第五人民医院肝病三科,河南安阳455000

出　　处：《肝脏》2021年第12期1339-1343,共5页Chinese Hepatology

摘　　要：目的探究miR-203a-3p对肝癌细胞迁移和侵袭的影响及其作用机制。方法体外培养HepG2细胞,将实验分为对照组(人肝癌细胞)、miRNA-NC组(转染阴性对照序列)和miR-203a-3p组(转染miR-203a-3p mimic)。Transwell和细胞划痕实验用以检测细胞侵袭和迁移能力;双荧光素酶实验用以验证miR-203a-3p和锌指转录因子(SLUG)的靶向关系;实时荧光定量聚合酶链式反应(qRT-PCR)用以检测miR-203a-3p和SLUG mRNA表达;蛋白质免疫印迹(WB)用以检测SLUG蛋白以及上皮间质转化相关蛋白E型钙黏蛋白(E-cadherin)、闭合蛋白(Occludin)、N型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)的表达;裸鼠成瘤实验用以检测转染后细胞体内成瘤能力。结果与对照组相比,miRNA-NC组细胞侵袭和迁移能力、miR-203a-3p的表达、SLUG mRNA和蛋白的表达、上皮间质转化相关蛋白的表达均差异无统计学意义(P>0.05);miR-203a-3p组细胞侵袭和迁移能力、SLUG mRNA和蛋白的表达、N-cadherin、Vimentin蛋白表达显著降低,miR-203a-3p的表达、E-cadherin、Occludin蛋白表达显著升高(P<0.05)。结论miR-203a-3p可能靶向调控SLUG,抑制上皮间质转化过程,从而抑制肝癌细胞迁移及侵袭。Objective To investigate the effect and mechanism of miR-203a-3p on the migration and invasion of hepatocellular carcinoma(HCC)cells.Methods HepG2 cells were cultured in vitro and divided into control group(human HCC cell line),miRNA-NC group(transfected with mimics control)and miR-203a-3p group(transfected with miR-203a-3p mimic).Transwell and scratch test were used to assess the invasion and migration ability.Luciferase reporter assay was used to verify the targeting relationship between miR-203a-3p and zinc finger transcription factor(SLUG).Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-203a-3p and SLUG mRNA.Western blot(WB)was used to detect the expression levels of SLUG protein,E-cadherin,Occludin,N-cadherin and Vimentin.The tumorigenicity ability of transfected cells was detected in nude mice.Results There was no significant difference in invasion and migration ability of cells between control group and miRNA-NC group,and the expression levels of miR-203a-3p,SLUG mRNA,SLUG protein and epithelial mesenchymal transition related protein between the two groups were not significantly different,(P>0.05).In miR-203a-3p group,the invasion and migration ability was significantly decreased.The expression levels of SLUG mRNA,SLUG protein,N-cadherin and Vimentin in miR-203a-3p group were significantly decreased,while the expression levels of miR-203a-3p,E-cadherin and Occludin were significantly increased(P<0.05).Conclusion MiR-203a-3p may inhibit the migration and invasion of HCC cells by inhibiting EMT via regulating SLUG.

关 键 词：miR-203a-3p 锌指转录因子 肝癌细胞 上皮间质转化 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]