肝细胞癌细胞株HepG2细胞甲胎蛋白基因沉默对Survivin表达的影响

Survivin mRNA affected in HepG2 cells of hepatocellular carcinoma after AFP gene silencing

目的 研究肝细胞癌细胞株HepG2细胞甲胎蛋白基因沉默（即基因不表达或表达量极低）对Survivin表达的影响.方法 应用siRNA技术对肝细胞癌细胞株HepG2细胞中甲胎蛋白基因的表达进行下调,应用ELISA法（即酶联免疫吸附测定法）对转染前后上清液甲胎蛋白的浓度进行测定,应用MTT比色法对转染前后细胞存活和生长情况进行检测,应用流式细胞术对细胞凋亡率进行分析,应用逆转录PCR技术对转染前后细胞Survivin mRNA水平进行检测.结果 与转染前相比,转染后研究组的上清液中甲胎蛋白的浓度均发生逐渐降低,且转染48 h后,观察组的下降更为明显.观察组的吸光度明显逐渐下降,肝癌细胞生长活性减弱了45.23％;HepG2细胞的凋亡率增至36.8％,较转染前升高了24.4％,HepG2细胞中Survivin mRNA表达下降了74.32％.对于上述情况,空白组和对照组均未见明显的变化.结论 肝细胞癌细胞株HepG2细胞甲胎蛋白基因沉默可抑制Survivin mRNA表达,从而降低肝癌细胞的生长活性以及增加细胞的凋亡率.

Objective To study the influence on Survivin expression in HepG2 cells of hepatocellular carcinoma after alpha-fetoprotein （AFP） gene silencing （ie gene is not expressed or is very low-volume expressed）.Methods AFP gene expression was down regulated in HepG2 cells by siRNA,and the content of AFP in the supcmatant was detected by ELISA.Survivin mRNA level was tested by RT-PCR.MTT was applied to evaluate cell proliferation.Flow cytometry was applied to evaluate cell apoptosis.Results Compared with previous transfection,the concentration of AFP in the supernatant of observation group gradually reduced after transfection,and at 48 hours after transfection,the degree of decrease in observation group was more obvious.The absorbance of observation group significantly decreased,cell proliferation activity decreased by 45.23%;apoptosis rate of HepG2 cells increased to 36.8% （by 24.4%）,and Survivin mRNA expression decreased by 74.32%.For above cases,there were no significant changes in blank group or control group.Conclusion HepG2 cells of hepatocellular carcinoma after AFP gene silencing can inhibit the mRNA expression of Survivin,so that it reduces the activity of liver cancer cells and increases apoptosis rate of liver cancer cells.