CyclinD1干扰RNA对肝癌细胞HepG2增殖和凋亡的作用

Effect of CyclinD1 Interference RNA on the Proliferation and Apoptosis in Hepatoma Cell Line HepG2

[目的]探讨干扰RNA对人肝癌细胞株HepG2的CyclinD1基因表达以及细胞增殖、凋亡的影响。[方法]将表达CyclinD1 siRNA的重组质粒pU6-CyclinD1-siRNA导入HepG2细胞,同时设立阴性对照空载体转染组及空白对照组。倒置荧光显微镜观察G418筛选稳定转染的细胞。MTT法检测细胞生长增殖率,流式细胞术检测细胞凋亡,RT-PCR、Western Blot方法检测CyclinD1沉默效果。[结果]与阴性对照组及空白对照组相比,转染组细胞生长速度减慢,凋亡率上升,CyclinD1 mRNA和蛋白表达水平均明显下降(P均<0.05)。[结论]RNA干扰技术可有效抑制细胞增殖,导致细胞凋亡,使CyclinD1 mRNA及蛋白表达水平均明显下降。

[Purpose ] To investigate the influence of CyclinD1 silenced on cell proliferation and apoptosis in human hepatoma cell line HepG2. [Methods] The recombinant plasmid pU6-Cy- clinDl-siRNA （CyclinD1 siRNA） was imported into HepG2 cells. Meanwhile the blank group and blank plasmid group were constructed. Stable transfeetion of cell was selected by G418. Rate of cell growth and proliferation was detected by MTI＇. Cell apoptosis was detected by flow cytometry. The expression of CyclinD1 was detected by RT-PCR and Western Blot. [Results] Compared with the blank group and blank plasmid group,in the stable transfeetion group ,the cell proliferation was slower, apoptosis rate was higher and the expression of CyclinD 1 was lower （P〈0.05）. [Conclusion] RNA interference technique can inhibit cell proliferation,induce cell apoptosis and downregulate expression of CyclinD1 mRNA and protein significantly.