RNA干扰survivin基因对HepG2细胞凋亡及化疗影响  

EFFECT OF RNA INTERFERENCE TARGETING AT SURVIVIN GENE ON APOPTOSIS AND CHEMOTHERAPY OF HEPATOCELLULAR CARCINOMA CELL LINE HepG2

作　　者：孙瑶[1] 雷炜[1] 

机构地区：[1]青岛大学医学院附属医院肿瘤科,山东青岛266003

出　　处：《齐鲁医学杂志》2010年第4期303-305,308,共4页Medical Journal of Qilu

摘　　要：目的探讨RNA干扰survivin基因对HepG2细胞凋亡及化疗敏感性的影响。方法构建针对HepG2细胞survivin基因的shRNA真核表达载体pshRNA-survivin-387。将HepG2细胞接种于6孔板中,分为空白对照组(不加药物)、阴性对照组(每孔加pshRNA-survivin-NC为3.2μg)、低浓度转染组(每孔加pshRNA-survivin-387为0.8μg)、中浓度转染组(每孔加pshRNA-survivin-387为1.6μg)、高浓度转染组(每孔加pshRNA-survivin-387为3.2μg),作用48 h后收集细胞,流式细胞术检测各组细胞凋亡指数;将HepG2细胞接种于96孔板中,分为3组:空白对照组(不加药物)、阴性对照组(每孔加pshRNA-survivin-NC为0.2μg)、实验组(每孔加pshR-NA-survivin-387为0.2μg),四甲基偶氮唑蓝法检测各组细胞分别与顺铂(2.0 mg/L)作用24、36、48、60 h后的增殖水平。结果各浓度转染组细胞的凋亡指数明显高于空白对照组和阴性对照组(F=228.87,q=10.45～38.21,P<0.01),高浓度转染组细胞凋亡指数高于低浓度转染组(q=23.99,P<0.01)。顺铂对实验组细胞的细胞抑制率在24、36、48、60 h时明显高于空白对照组和阴性对照组(F=235.88～910.86,q=28.04～52.28,P<0.01)。结论 pshRNA-survivin-387转染能诱导肝癌细胞的凋亡,增强肝癌细胞对顺铂化疗的敏感性。Objective To investigate the effect of RNA interference targeting at survivin gene on the apoptosis and chemosensitivity of hepatocellular carcinoma cell line HepG2. Methods Survivin gene sequence specific shRNA vectors pshRNA-survivin-387 were constructed.HepG2 cells cultured in 6-well culture dish were divided into five groups： blank control group（without any drug）,negative control group（3.2 μg pshRNA-survivin-NC 3/well）,low-dose group（0.8 μg pshRNA-survivin-387/well）,medium-dose group（1.6 μg pshRNA-survivin-387/well） and high-dose group（3.2 μg pshRNA-survivin-387/well）.After 48 h of transfection,cultured cells were harvested,and the apoptotic index was examined by flow cytometry.HepG2 cells cultured in 96-well culture dish were divided into three groups：blank control group（without any drug）,negative control group（0.2 μg pshRNA-survivin-NC/well）,and pshRNA-survivin-387 group（0.2 μg pshRNA-survivin-387/well）.MTT assay was used to detect the rate of cellular inhibition effected by C-DDP（2.0 mg/L） after 24,36,48 and 60 hours. Results The apoptotic index of the low-,medium-and high-dose pshRNA-survivin-387 groups were significantly higher than those of the blank control group（F=228.87,q=14.21-38.21,P0.01） and those of the negative control group（q=10.45-34.40,P0.01）.Apoptotic index of high-dose pshRNA-survivin-387 groups were significantly higher than that of the low-dose pshRNA-survivin-387 groups（q=23.99,P0.01）.The rate of inhibition of C-DDP in pshRNA-survivin-387 groups were significantly higher than those of the control groups（F=235.88-910.86,q=28.04-52.28,P0.01）. Conclusion Transfection of the pshRNA-survivin-387 to HepG2 cell can induce the apoptosis of liver cancer cells,and increase the sensitivity of the cells to chemotherapy.

关 键 词：肝肿瘤 SURVIVIN RNA干扰 药物疗法 

分 类 号：R730.54[医药卫生—肿瘤]