saRNA调控p21基因表达对肝癌细胞的抑制作用

Study of targeting gene p21 by saRNA inhibits hepatocellular carcinoma cells

目的探讨RNA激活p21对肝癌Hep G2、Hep 3b和SMMC-7721细胞生长和侵袭力的影响。方法化学合成靶向p21的saRNA,将其转染肝癌细胞系Hep G2、Hep 3b和SMMC-7721,利用荧光定量RT-PCR和免疫印迹检测细胞中的p21表达水平,并MTT法检测细胞生长情况及划痕实验观察细胞侵袭能力的变化。结果 Hep G2、Hep3b和SMMC-7721细胞的p21 mRNA分别较对照组升高了6.6、7.1、6.1倍,p21蛋白表达水平分别升高了17.4、11.4、6.7倍。细胞转染后48 h时的细胞增长平均抑制率分别为47%、53%和55%;划痕实验显示转染组细胞迁移能力显著低于对照组(P<0.01)。结论靶向p21的RNAa能抑制肝癌细胞的生长和侵袭力,p21可作为一个具有肝癌治疗应用价值的靶基因。

Objective To explore the effect of up- regulating expression p21 induced by saran on proliferation and invasion in hepatocellular carcinoma Hep G2,Hep 3b,SMMC- 7721 cells. Methods A saRNA,targeting gene p21 was synthesized. Hep G2,Hep 3b,SMMC- 7721 cells were cultured in vitro and transfected with saRNA,then RT- PCR and Western blotting were applied to detect the expression level of p21 mRNA and protein,respectively. The proliferation of the cells were analyzed by MTT kits. The invasion power of the cells were detected by scratch test. Results The p21 mRNA of Hep G2,Hep 3b,SMMC- 7721 cells were higher than the control group of 6. 6,7. 1,6. 1 times,respectively.And the p21 protein level were 17. 4,11. 4,6. 7 times,respectively. The three cells average inhibition rate were 47%,53% and 55%,respectively. In contrast to the blank control group,the cell migration activity of transfection group was declined（ P〈0. 01）. Conclusion RNAa targeting p21 can inhibit the proliferation and invasion power of Hep G2,Hep 3b,SMMC- 7721 cells. The p21 may be a target point of gene therapy for hepatocellular carcinoma.