端粒酶反义寡聚脱氧核苷酸对人肝癌细胞的作用

The effect of telomerase antisense oligodeoxynucleotides on the growth of hepatoma cell line

目的 观察端粒酶反义寡聚脱氧核苷酸对人肝癌细胞株的作用。方法 用端粒酶反义寡聚脱氧核苷酸与人肝癌细胞株共同温育一定时间后 ,观察细胞形态变化 ,检测细胞DNA含量的分布 ,测定端粒酶活性。结果 端粒酶反义寡聚脱氧核苷酸能诱导人肝癌细胞凋亡 ,抑制端粒酶活性 ,在形态学上表现为细胞膜起泡、染色质固缩、核碎裂、凋亡小体形成 ;电泳呈凋亡特征性Ladder带 ;流式细胞仪分析显示 ,在G1 期前出现亚二倍体凋亡峰 ;端粒酶活性抑制。结论 端粒酶反义寡聚脱氧核苷酸能诱导肝癌细胞凋亡 ,抑制端粒酶活性 ,抑制端粒合成 。

Objective To observe the effect of telomerase antisense oligodeoxynucleotides on hepatoma cell line. Methods Hepatoma cancer cell line were cultured with 1～5μmol/L telomerase antisense oligo deoxynucleotides for various duration, morphological changes and cell DNA disribution were studied. The apoptotic cell were masured with flow cytometer. Telomerase activity was detected by TRAP ELISA. Results Hepatoma cells cultured with 5μmol/L telomerase antisense oligodeoxynucleotides presented apoptotic features: bulbing cell membrance, chromatin condensation, nucleic fragmentation and apoptotic body formation. Flow cytometer analysis showed a sub G 1 cell peak; agarose electrophoresis showed marked DNA ladder. These results illustrated that the telomerase activity was inhibited. Conclusion Telomerase may play a crucial role in the growth of hepatoma cells. Telomerse antisense oligodeoxynucleotides may inhibit telomerase activity. It may be a new target of treatment of hepatoma.