下调微小RNA-1表达对人肝癌血管内皮细胞生物学行为的影响  

Effects of suppressing the expression of microRNA-1 on biological behaviors in the vascular endothelial cells of human hepatocellular carcinoma

作　　者：胡超[1] 沈世强[1] 崔忠惠[1] 

机构地区：[1]武汉大学人民医院肝胆外科,430060

出　　处：《中华实验外科杂志》2014年第10期2108-2111,共4页Chinese Journal of Experimental Surgery

基　　金：武汉市科技攻关计划资助项目（201260523178-2）

摘　　要：目的 通过构建慢病毒介导的微小RNA（miRNA）-1-小干扰RAN （siRNA）,观察下调miR-1表达对人肝癌血管内皮细胞（TEC）生物学行为的影响.方法 针对目标序列合成特异性siRNA干扰片段并克隆入慢病毒载体,将重组miRNA-1-siRNA-GV159转染TEC,实验分为3组：（1）下调组（miRNA-1-siRNA,MD组）;（2）阴性对照组（NC组）;（3）空白对照组（CON组）.分别用噻唑蓝（MTT）法检测细胞增殖,流式细胞仪检测细胞的凋亡及细胞周期,划痕实验检测细胞的迁移能力.结果 MTT结果显示MD组细胞在转染miRNA-1-siRNA-GV159后的第1、2、3、4、5天5个时间点均表现出吸光度值降低,与NC组及CON组比较差异有统计学意义（P＜0.05）;流式细胞仪检测凋亡结果显示CON、NC、MD组凋亡率分别为（2.03±0.30）％、（2.18±0.15）％、（6.32±0.33）％,MD组细胞凋亡率比其他两组明显明显增加,差异有统计学意义（P＜0.05）;而细胞周期检测结果显示3组之间差异无统计学意义（P＞0.05）;划痕实验结果显示划痕12h后CON、NC、MD3组细胞的迁移距离分别为（270.6 ±44.5）、（299.5 ±28.7）、（198.4 ±49.6）μm,MD组和其他两组比较迁移能力明显下降,差异有统计学意义（P＜0.05）.结论 下调miRNA-1表达可抑制TEC的增殖能力,增加TEC的凋亡并降低其迁移能力.Objective To study the effects of suppressing the expression of microRNA （miRNA）-1 by lentivirus-mediated miRNA-1-siRNA on biological behaviors in the vascular endothelial cells of human hepatocellular carcinoma （TEC）.Methods miRNA-1 specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector.TEC were infected by miRNA-1-siRNA recombinant lentivirus （miRNA-1-siRNA-GV159）.TEC cells were divided into miRNA-1 down-regulation （MD） group,negative control （NC） group,control （CON） group.The proliferation of TEC were detected by thiazolyl blue （MTT） assay,the cell cycle and the apoptosis of TEC were detected by flow cytometry,and the cell migration was assessed by a wound healing assay.Results The results of MTT showed that at 1,2,3,4,5 d after infected by miRNA-1-siRNA-GV159 the optical density of TEC in the MD group were all significantly lower than the other two groups （P 〈 0.05）.The results of flow eytometry showed that the apoptosis rate of TEC in CON group,NC group,MD group was （2.03 ±0.30）%,（2.18±0.15）%,（6.32 ±0.33）% respectively.The apoptosis rate of TEC in MD group was significantly increased compared with the other two group （P 〈 0.05）.However,there was no obvious differences among the there groups on the cell cycle of TEC after suppressing the expression of miRNA-1 （P 〉 0.05）.The results of wound healing assay showed that 12 h after scratching the distance of migration of TEC in CON group,NC group,MD group was （270.6 ± 44.5）,（299.5 ± 28.7）,（198.4 ± 49.6） μm respectively.The distance of migration in MD group was significantly decreased conpared with the other groups （P 〈 0.05）.Conclusion Suppressing the expression of miRNA-1 can suppress the proliferation,increase the apoptosis and decrease the migration of TEC.

关 键 词：肝细胞肝癌 肝癌血管内皮细胞 微小RNA-1 RNA干扰 

分 类 号：R735.7[医药卫生—肿瘤]