人端粒酶催化亚基反义核酸对子宫内膜癌细胞的抑制作用

Antisense oligodeoxynucleotides of human telomerase reverse transcriptase inhibit endometrial carcinoma cell HEC 1A proliferation

目的 探讨人端粒酶催化亚基 (hTERT)基因反义寡核苷酸 (AODN)对子宫内膜癌细胞系HEC 1A的生长抑制作用及作用机理。方法 采用四甲基偶氮唑蓝 (MTT)法检测AODN对细胞增殖能力及细胞生长的影响 ;逆转录聚合酶链反应技术 (RT PCR)、定量端粒酶重复扩增法 (TRAP)、Westernblot蛋白免疫印迹法和凋亡相关酶活性测定检测反义核酸对hTERT基因转录、蛋白表达、细胞端粒酶活性以及凋亡相关酶活性的变化。结果 低浓度hTERT基因AODN能够下调HEC 1A细胞hTERTmRNA含量 ,抑制细胞hTERT蛋白表达 ,下调端粒酶活性 ,并且激活凋亡相关酶Caspase 1和Caspase 3活性 ;其对细胞的生长抑制作用有明显的时效性和剂量依赖性。结论 hTERT基因反义核酸能够抑制子宫内膜癌细胞的增殖能力 ,有望成为内膜癌治疗的新方向。

Objective To evaluate antisense technology for human telomerase inhibition in the treatment of endometrial cancer. Methods An antisense oligodeoxynucleotides (AODN) directed against the human telomerase transcriptase (hTERT), designed and synthesized to serve as a telomerase inhibitor, was tansfected into endometrial carcinoma cell line HEC 1A by lipofectin. Reverse trascription polymerase chain reaction (RT PCR) and Western blot were used to test the expression of hTERT mRNA and hTERT protein before and after transfection. Telomerase activity was tested by telomeric repeat amplification protocol. The proliferation and growth of HEC 1A were also studied by methyl thiazolyl tetrazolium and cell growth curve before and after tansfection. Results AODN could down regulate the expression of hTERT mRNA and protein, inhibiting telomerase activity and proliferation of endometrial cancer cell line in a dose and period dependent manner. Conclusion Antisense oligodeoxynucleotides of human telomerase transcriptase definitely inhibits the proliferation of endometrial cancer cell line. Telomerase inhibitor may thus become a new gene therapeutic agent for endometrial carcinoma. [