蟾蜍灵通过抑制端粒酶活性诱导人脑胶质瘤细胞死亡

Bufalin inhibits the activity of human telomerase reverse transcriptase and induces human glioma cell death

目的:探讨蟾蜍灵对人脑胶质瘤的作用。方法:将不同浓度的蟾蜍灵作用于胶质瘤细胞后,用CCK-8法检测细胞增殖抑制率,克隆形成实验检测细胞法检测增殖抑制,Western-blot测定人端粒酶逆转录酶(hTERT)的蛋白表达,Real time-PCR检测hTERT基因表达水平,CCK-8法检测蟾蜍灵对人原代胶质瘤细胞的杀伤作用。结果:蟾蜍灵对胶质瘤细胞株细胞U251、U87增殖具有抑制作用,且呈时间和剂量依赖性,并且能够有效抑制U251及U87细胞的克隆形成。Western-blot结果显示hTERT蛋白表达随药物浓度增加而递减,Real time-PCR结果证实蟾蜍灵降低了hTERT基因表达。同时体外实验证明,蟾蜍灵对人原代胶质瘤细胞具有明显杀伤作用。结论:蟾蜍灵能够抑制胶质瘤细胞系细胞及人原代胶质瘤细胞增殖,同时通过抑制hTERT表达降低端粒酶活性以发挥其抗肿瘤的作用。

Objective：To investigate the effect of Bufalin on human glioma.Methods：U87 and U251 cells were treated with different concentrations of Bufalin and cell proliferation inhibition rate was measured by using CCK-8 assay.Clone formations were counted by staining with Giemsa.The expression of hTERT （human telomerase reverse transcriptase） protein was detected by Western-blot.Real time-PCR was performed to detect the gene expression of hTERT.Moreover,patient derived glioma cells were treated with Bufalin,and cell viabilities were measured by CCK-8 assay.Results：Bufalin suppressed cell growth and colony formation in U87 and U251 cells.Treatment with Bufalin inhibited hTERT gene expression and was consequent on the decrease of hTERT protein expression.Furthermore,Bufalin displayed inhibitory effects on patient derived glioma cells.Conclusion：Taken together,Bufalin can suppress glioma cell viability by inhibiting hTERT activity and may act as an effective reagent for glioma treatment.