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目的观察在抑制肝癌细胞端粒酶活性表达情况下,1,25-二羟基维生素D3(1,25-VD3)对肝癌细胞凋亡的影响。方法经脂质体介导,将反义端粒酶RNA真核表达载体pBBS212/hTR导入维生素D3受体(VDR)阳性的肝癌细胞株SMMC7721细胞中培养。细胞克隆转移扩大培养,将其分为对照组、药物组、转染组和转染药物组。添加1,25-VD3,分别作用于转染药物组、药物组,用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)和电子显微镜检测肝癌细胞凋亡。结果转染组和对照组细胞的端粒酶活性分别为0.686和1.685,说明反义端粒酶RNA可显著降低肝癌细胞端粒酶活性。对照组、药物组、转染组和转染药物组细胞凋亡率分别为3.4%、12.2%、8.8%、23.6%;差异有显著统计学意义(P<0.01)。超微结构检查也证实存在凋亡发生。1,25-VD3或转染反义端粒酶RNA对肝癌细胞有促细胞凋亡作用,两者协同对肝癌细胞凋亡作用显著增强。结论转染反义端粒酶RNA,可降低肝癌细胞端粒酶活性,可显著增强1,25-VD3对肝癌细胞的促凋亡作用,并且对细胞的增殖也存在明显的抑制作用。
Objective To investigate the effect of 1,25-dihydroxyvitamin D3 (1,25-VD3) on the apoptosis of hepatocellular carcinoma cells in the presence of inhibition of telomerase activity. Methods The liposome-mediated antisense telomerase RNA eukaryotic expression vector pBBS212 / hTR was transfected into VDR-positive hepatoma cell line SMMC7721. Cell cloning and metastasis expansion culture, divided into control group, drug group, transfection group and transfection drug group. 1,25-VD3 was added to the transfection drug group and drug group, respectively. The apoptosis of hepatocellular carcinoma cells was detected by TUNEL and electron microscopy. Results The telomerase activity of transfected and control cells were 0.686 and 1.685, respectively, indicating that antisense telomerase RNA can significantly reduce the telomerase activity of hepatoma cells. The apoptosis rates of control group, drug group, transfection group and transfection group were 3.4%, 12.2%, 8.8% and 23.6%, respectively. The difference was statistically significant (P <0.01). Ultrastructural examination also confirmed the presence of apoptosis. 1,25-VD3 or transfected antisense telomerase RNA on human hepatocellular carcinoma cell apoptosis, both synergistic effect on the apoptosis of hepatoma cells was significantly enhanced. CONCLUSION: Transfection of antisense telomerase RNA can reduce the telomerase activity of hepatocellular carcinoma cells and significantly enhance the pro-apoptotic effect of 1,25-VD3 on hepatocellular carcinoma cells, and also inhibit the proliferation of hepatoma cells.