5型腺病毒E1A基因通过调控GP73的表达抑制肝癌细胞增殖的初步研究

Adenovirus Type 5 E1A Gene Inhibits Proliferation of HCC Cell Lines by Regulating GP73 Expression

目的:研究5型腺病毒(Ad5)E1A基因通过调控高尔基蛋白73(GP73)的表达水平,抑制肝癌细胞HepG2的增殖。方法:以腺病毒为模板、pcDNA-flag-Vector为载体,构建真核表达载体pcDNA3-Flag-Ad5E1A;免疫印迹实验鉴定重组蛋白Ad5E1A的表达;免疫印迹实验验证Ad5E1A对GP73表达水平的影响;将Ad5E1A基因转染HepG2细胞,用CCK-8试剂盒检测HepG2细胞的增殖情况。结果:免疫印迹实验结果证实,真核表达载体pcDNA3-Flag-Ad5E1A在HEK293细胞中得到表达;Ad5E1A可明显抑制GP73的表达。酶标仪检测发现,与转染Flag-GP73组相比,Ad5E1A组的抑制率为17.5%,差异无统计学意义(P>0.05);而与转染Flag-GP73组比,共转Ad5E1A和GP73组的抑制率为37.8%,差异有统计学意义(P<0.05)。结论:Ad5E1A基因通过调控GP73的表达抑制了肝癌细胞HepG2的增殖。为研究肝癌发生机制,开发新型治疗方案提供了实验基础。

Objective:To investigate the adenovirus type 5(Ad5)E1A gene inhibits proliferation of hepatocellular carcinoma(HCC)cell lines by regulating the expression of Golgi protein 73(GP73).Methods:Ad5E1A was cloned from adenovirus and inserted into pcDNA-flag-Vector to construct pcDNA3-Flag-Ad5E1A.Western blotting was used to detect the expression of pcDNA3-Flag-Ad5E1A and detect the effect of Ad5E1A on the expression of GP73.After HepG2 cells were transfected Ad5E1A gene,the proliferation of HepG2 cells was detected by CCK-8 assay kit.Results:Western blotting showed that Flag-Ad5E1A was efficiently expressed in HEK293 cells.The expression of GP73 was inhibited with Ad5E1A gene transfection.The results showed that the inhibition rate of the Ad5E1A group was 17.5%compared with Flag-GP73 group(P>0.05),the inhibition rate of Ad5E1A and GP73 co-transfected group was 37.8%compared with Flag-GP73 group(P<0.05).Conclusion:The Ad5E1A gene inhibits proliferation of HCC cell lines by regulating the expression of GP73.It provides experimental basis for investigating the mechanism of hepatocellular carcinoma and developing new therapeutic strategies.