MicroRNA-30a-5p对肝癌细胞株SMMC-7721增殖凋亡及侵袭转移的影响  

Effect of miR-30a-5p on the proliferation, apoptosis, invasion and migration of SMCC-7721 human hepatocellular carcinoma cells

作　　者：代航[1] 康冰[1] 左的于[2] 左国庆[3] 

机构地区：[1]重庆医科大学附属第二医院消化内科,400010 [2]重庆医科大学第一临床学院 [3]重庆市中医院

出　　处：《中华肝脏病杂志》2014年第12期915-920,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金（81271598）;重庆市自然科学基金（2012-1-035）

摘　　要：目的 探讨转染miRNA-30a-5p对肝癌细胞生物学行为的影响. 方法 将miRNA-30a-5p模拟物、miRNA-30a-5p抑制物瞬时转染入肝细胞肝癌细胞株SMCC-7721,应用实时荧光定量PCR检测正常肝细胞株L02及肝癌细胞株SMCC-7721转染后的miR-30a-5p的mRNA表达情况,CCK-8法检测细胞增殖能力,克隆形成实验检测集落形成情况,流式细胞术检测细胞凋亡及各组细胞周期分布的差异,Transwell小室检测各组细胞体外侵袭转移能力,建立BALB/c-nu裸小鼠肝癌模型并观察miRNA-30a-5p对肿瘤生长的影响. 结果 实时荧光定量PCR显示,与未转染组及正常肝细胞株L02相比,肝癌细胞株SMCC-7721在转染miRNA-30a-5p模拟物后,mRNA表达明显上调（P＜0.01）,而转染miRNA-30a-5p抑制物的mRNA表达明显受抑（P＜0.01）,差异有统计学意义.肝细胞肝癌细胞株SMCC-7721在转染miRNA-30a-5p模拟物后,细胞活性、克隆形成能力、迁移和侵袭能力与miRNA-30a-5p抑制物转染组、未转染组及正常肝细胞株L02相比,相对减弱（P＜0.05）,miRNA-30a-5p模拟物转染组凋亡率,高于miRNA-30a-5p抑制物转染组、未转染组及正常肝细胞株L02 （P＜0.05）,细胞周期出现S期阻滞.裸鼠肝癌模型中,实验组裸鼠瘤体质量及体积明显小于空载体对照组和空白对照组（P＜ 0.05）.结论 上调miR-30a-5p表达可明显抑制肝细胞肝癌细胞株SMCC-7721的增殖,促进其凋亡,抑制其迁移、侵袭能力,并且抑制裸小鼠肝癌模型肿瘤的生长.Objective To explore the effect ofmicroRNA-30a-5p （miRNA-30a-5p） on the biological behavior of human hepatoma cells.Methods The liver cancer cell line SMCC-7721 cells and the normal liver cell line L02 cells （control） were transiently transfected with miRNA-30a-5p mimics and an miRNA-30a-5p inhibitor by Lipofectamine 2000 （Life Technologies）.miR-30a-5p mRNA expression was detected by quantitative real-time （q）PCR.Cell proliferation was evaluated using the Cell Counting Kit-8 （CCK-8） assay and apoptosis was assessed by flow cytometry.Invasion and migration were measured by transwell chamber assays.The SMCC-7721 cells was injected subcutaneously into nude mice to establish a tumor animal model.Results The SMCC-7721 cells transfected with miRNA-30a-5p mimics showed significantly higher miRNA-30a-5p mRNA expression than the non-transfected SMCC-7721 cells and the transfected control L02 cells （P 〈 0.01）.The miRNA-30a-5p mRNA expression was significantly lower in the SMCC-7721 cells transfected with the miRNA-30a-5p inhibitor than the non-transfected SMCC-7721cells the control L02 cells （P 〈 0.01）.The overexpression of miRNA-30a-5p inhibited the viability,colony formation rate,and invasion and migration abilities,as shown in the cells transfected with the miRNA-30a-5p mimics （P 〈 0.05）; in addition,the miRNA-30a-5p promoted proliferation of cells （P 〈 0.05）,as shown by more S phase cells detected by flow cytometry.SMCC-7122 cells transfected with miRNA-30a-5p mimics produced tumors with significantly higher average weight than tumors produced by SMCC-7122 cells that were untransfected or transfected with empty vector （bothP 〈 0.01）.Conclusion Overexpression ofmiR-30a-5p had an inhibitory effect on cell proliferation,induced apoptosis,increased the number of cells in S phase,and markedly inhibited invasion and migration of SMCC-7721 HCC cells in vitro and in vivo.

关 键 词：癌 肝细胞 微小RNA 增殖 凋亡 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]