抑制端粒酶活性对1,25-(OH)_2D_3诱导肝癌细胞增殖分化的影响

Effect of the down-regulating telomerase activity on the proliferation and differentiation of hepatocarcinoma cells induced by 1,25-dihydroxyvitamin D_3

目的:观察抑制端粒酶活性对1,25-(OH)2D3诱导肝癌细胞增殖分化的影响作用。方法:经脂质体介导,将反义端粒酶RNA转染肝癌细胞系SMMC7721细胞。转染细胞扩大培养后,添加1,25-(OH)2D3分别作用于各实验组细胞后,采用MTT法、平板克隆形成实验检测细胞的存活和生长;光镜和电子显微镜检测细胞分化形态改变。结果:对照组与转染组的吸光度值分别为1.685和0.686,说明转染反义端粒酶RNA可显著抑制肝癌细胞的端粒酶活性;而在抑制肝癌细胞端粒酶活性的基础上,1,25-(OH)2D3抑制肝癌细胞增殖和诱导细胞分化的作用得以显著的增强。形态学检查也进一步证实存在诱导分化改变。结论:在降低细胞端粒酶活性的基础上,1,25-(OH)2D3抑制肝癌细胞增殖、诱导细胞分化的作用得到增强。

Objective：To observe the effect of down-regulating telomerase activity on proliferation and differentiation of hepatocarcinoma cells induced by 1,25-dihydroxyvitamin D3.Methods： The antisense telomerase RNA gene was introduced into SMMC7721 cells by using lipofectin mediation.After the transfected cells were cultured enlargely,1,25-dihydroxyvitamin D3 was added into the experimental group.Then MTT assay and clone formation assay were applied to detect the cells survival and growth.Meanwhile the light microscope and electron microscope were used to detect phenotypes and differentiation of cells.Results：The absorbance of the transfected cells and control group were 0.686 and 1.685,respectively.The results suggests that antisense telomerase RNA had a significant inhibition of telomerase activity of hepatocarcinoma cells.The inhibition of cell growth,reduction of colony-forming rate,and induction of cell differentiation induced by 1,25-dihydroxyvitamin D3 was significantly enhanced after the expression of telomerase activity blocked by transfected antisense elomerase RNA in SMMC7721 cells.Conclusion：Inhibition of telomerase activity could significantly enhance cell apoptosis,inhibit cell proliferation,and induce cell differentiation induced by 1,25-dihydroxyvitamin D3 in hepatocarcinoma cells.