RNAi抑制高迁移率族蛋白B2表达对人肝癌细胞HepG2迁移及侵袭能力的影响及机制  

Effect of RNAi inhibiting high mobility group box 2 expression on the migration and invasion of HepG2 cells and its mechanism

作　　者：魏雁虹[1] 杨晨雪 杨广民[1] 宋帅[1] 李明[1] 杨海娇 魏海峰[2] WEI Yanhong;YANG Chenxue;YANG Guangmin;SONG Shuai;LI Ming;YANG Haijiao;WEI Haifeng(Medical Laboratory Center,Jilin Province People's Hospital,Jilin Province,Changchun130021,China;Central Laboratory,Jilin Province People's Hospital,Jilin Province,Changchun130021,China)

机构地区：[1]吉林省人民医院医学检验中心,吉林长春130021 [2]吉林省人民医院生物治疗与基因诊断重点实验室,吉林长春130021

出　　处：《中国当代医药》2023年第31期11-14,F0003,共5页China Modern Medicine

基　　金：吉林省科技发展计划项目(20200201346JC)。

摘　　要：目的采用RNA干扰技术下调肝癌细胞中高迁移率族蛋白B2(HMGB2)的表达,观察其对肝癌细胞迁移及侵袭能力的影响,并阐明其作用机制。方法体外培养人肝癌细胞HepG2,分为HMGB2 RNA干扰组(HMGB2 siRNA)及阴性对照组(NC),分别以脂质体2000为载体,转染敲除HMGB2序列的RNA寡核苷酸(RNA oligo)及RNA干扰无关序列。采用RT-qPCR及Western blot法检测细胞中HMGB2 mRNA及蛋白表达水平以验证干扰效果;采用细胞划痕实验、Transwell侵袭实验检测敲低HMGB2对肝癌细胞迁移及侵袭能力的影响;Western blot法检测敲低HMGB2对HepG2细胞EMT相关蛋白表达的影响。结果HMGB2 siRNA组细胞HMGB2 mRNA及蛋白表达水平低于NC组,差异有统计学意义(P<0.05),表明RNA干扰成功;细胞划痕及Transwell侵袭实验结果显示,HMGB2 siRNA组细胞愈合率及侵入小室的细胞数量少于NC组,差异有统计学意义(P<0.05),表明下调HMGB2表达可降低HepG2细胞的迁移及侵袭能力;Western blot结果显示,HMGB2 siRNA组HepG2细胞N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)蛋白表达水平低于NC组,E-钙黏蛋白(E-cadherin)表达水平高于NC组,差异有统计学意义(P<0.05)。结论下调HMGB2表达可抑制肝癌细胞的上皮间质转化进程,进而降低其迁移及侵袭能力。Objective To observe the effect of down-regulation of high mobility group box 2(HMGB2)by RNA interference technology on the migration and invasion ability of hepatocellular carcinoma cells,and clarify its mechanism.Methods HepG2 cells were cultured in vitro and divided into normal control group(NC group)and HMGB2 small interference RNA group(HMGB2 siRNA group).The RNA oligonucleotides(RNA oligonucleotides)and unrelated RNA oligonucleotides were transfected in HepG2 cells with Lipofectamin 2000 respectively.To verify the interference effect,RT-qPCR and Western blot were used to detect the expression level of HMGB2 mRNA and protein in cells.The migration and invasion abilities of hepatoma cells were detected by cell scratch test and Transwell invasion test respectively.The expressions of EMT-related protein in HepG2 cells were detected by western blot method.Results The expression of HMGB2 mRNA and protein in HMGB2 siRNA group were lower than those in NC group,and the differences were statistically significant(P<0.05),indicating that RNA interference was successful.The results of cell scratch and Transwell invasion test showed that cell healing rate and the number of cells invading the cell compartment in HMGB2 siRNA group was less than NC group,and the difference was statistically significant(P<0.05),indicating that down-regulation of HMGB2 expression could reduce the migration and invasion ability of HepG2 cells.Western blot results showed that the expression of N-cadherin and Vimentin in HepG2 cells in HMGB2 siRNA group were lower than those in NC group,and the expression of E-cadherin was higher than that in NC group,the differences were statistically significant(P<0.05).Conclusion Down-regulation of HMGB2 expression can inhibit the process of epithelial-mesenchymal transformation of hepatocellular carcinoma cells,thereby reducing their migration and invasion ability.

关 键 词：肝癌 迁移 侵袭 高迁移率族蛋白B2 上皮间质转化 

分 类 号：R735.7[医药卫生—肿瘤]