短发卡状RNA抑制AKT2对肝癌细胞增殖及侵袭的影响  

The effects of inhibiting AKT2 by siRNA on the proliferation and invasion of liver cancer cells

作　　者：谢毅[1] 王旺河[1] 张超[1] 李国庆[1] 田鹏[1] 张辉[1] 王志凯[1] 梁鸿[1] 钱海鑫[2] 

机构地区：[1]河南省人民医院腹腔镜治疗研究中心,郑州450003 [2]苏州大学附属第一医院

出　　处：《中华肝胆外科杂志》2012年第10期784-787,共4页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的探讨短发卡状RNA（siRNA）抑制AKT2对肝癌细胞增殖、凋亡和侵袭转移的影响。方法设计并合成特异性靶向AKT2的siRNA片段并构建sMMc7721AKT2-siRNA表达质粒，将其转染SMMC7721细胞，通过G418筛选出稳定株。MTT法检测肝癌SMMC7721细胞的生存率变化；流式细胞术检测细胞周期；Western-blot检测P27、CyclinD1；Transwell实验和划痕实验分析细胞侵袭、转移能力的改变。结果MTT检测显示AKT2干扰可抑制sMMc7721细胞的生长，与其他组比较差异有统计学意义（P〈0．05）。流式细胞术显示AKT2干扰组细胞周期阻滞于G1期，G1期细胞比例上升，s期细胞比例下降。Western-blot检测显示CyclinD1表达下降，P27的表达上升。Transwell试验和划痕试验显示AKT2干扰组的侵袭和转移能力受到抑制。结论AKT2基因沉默可明显抑制肝癌SMMC7721细胞的生长并阻滞细胞周期。AKT2基因沉默可抑制肝癌SMMC7721细胞的侵袭和转移能力。Objective To study the effects of inhibing AKT2 by siRNA on SMMC7721 liver cancer cells proliferation, apoptosis,migration and invasion. Methods The siRNA targeting AKT2 was designed and the SMMC7721AKT2-siRNA plasmid was constructed and transfected into SMMC7721 cells. The stable cell lines were screened by G418. The effects of AKT2 by siRNA on SMMC7721 liver cancer cells, growth inhibition was evaluated by MTT assay. Cell cycle was analyzed by flow cytometry （FCM）. Protein of P27 and CyclinD1 was evaluated by Western-blot. The ability of migration and invasion was evaluated by wound healing and Transwell assay. Results The growth of SMMC7721 cells was significantly inhibited by siRNA （P〈0.05）. Flow cytometry display that AKT2 by siRNA can induce G1 phase arrest, the ratio of G1 phase increased homologously and S phase de- clined homologously. The protein of CyclinD1 was declined and the protein of P27 was increased by Western-blot. Wound healing and Transwell assay show that the ability of cells, migration and inva- sion was inhibited by AKT2 by siRNA. Conclusion AKT2 by siRNA can significantly inhibit the growth of SMMC7721 cells, arrest cell cycle. AKT2 by siRNA can inhibit the ability of invasion and migration of SMMC7721 cells.

关 键 词：肝癌 细胞 基因 AKT2 RNA干扰 侵袭 转移 

分 类 号：R735.7[医药卫生—肿瘤]