RNA干扰对过表达Hlrg的Hep G2细胞的影响

为观察RNA干扰对过表达Hlrg蛋白的Hep G2细胞的影响,构建了封闭Hlrg基因表达的shRNA表达载体pAVU6+27-Hlrg,将pAVU6+27-Hlrg稳定转染到过表达Hlrg蛋白的Hep G2细胞中;分别用相差倒置显微镜、FCM、SABC-FITC、RT-PCR、Western blot等观察细胞改变。测序结果表明pAVU6+27-Hlrg构建成功;针对Hlrg的RNA干扰已完成;相差倒置显微镜发现细胞突起增加;FCM显示干扰组G1期明显缩短。针对Hlrg的RNA干扰能促进过表达Hlrg蛋白的Hep G2细胞的生长。

Influence of RNA interference targeting againt over-expression of human lipopolysaccharide responsive gene on Hep G2 cells

The influence of RNA interference targeting on the over-expression of human lipopolysaccharide response gene(Hlrg) on Hep G2 cells was investigated,in which the expression vector pAVU6( )27-Hlrg that could block the expression of the Hlrg gene was constructed,and subcloned into shRNA vector pAVU6( )27.Then the pAVU6( )27-Hlrg plasmid was trasfected to Hep G2-pcDNA3.1( )-Hlrg cell line by means of trasfection reagent.The changes in the trasfected cells were examined by contrast invert microscopy,FCM,SABC-FITC,RT-PCR and Western blotting analysis.It was demonstrated that the plasmid pAVU6( )27-Hlrg was successfully constructed and the RNA interference targeting againt Hlrg had been completed.As demonstrated by invert microscopy,the cellular processes increased and the FCM analysis revealed the remarkable shortening of the G1 phase in the RNA interference group.It is concluded the RNA interference targeting can improve the growth and proliferation of Hep G2 cells in which the human lipopolysaccharide response protein is over-expressed.