端粒酶反义寡核苷酸对肾癌细胞端粒酶活性及其体外增殖的影响

Antisence oligonucleotides of telomerase in the inhibition of telomerase activity and renal carcinoma cell proliferation in vitro

目的 探讨端粒酶反义核酸对肾癌细胞端粒酶活性和体外增殖的影响。 方法 以端粒酶RNA模板区为靶点 ,序列为TAGGGTTAGACAA的硫代磷酸修饰的反义寡核苷酸 (ASON )与肾癌细胞株RCZ细胞共同孵育 ,计 1～ 2 8天细胞数 ,MTT比色法测细胞生长抑制率 ,PCR ELISA法检测端粒酶活性。 结果 ASON实验组与随机引物及空白对照组比较 ,端粒酶活性明显降低 (P<0 .0 0 1) ,细胞增殖数目显著减少 (P <0 .0 1) ,抑制率显著增加 (P <0 .0 0 1) ,抑制效果显示剂量依赖性 (P <0 .0 5 ) ,具有序列选择的特异性。 结论 以端粒酶RNA模板区为靶点的反义寡核苷酸抑制肾癌RCZ细胞的端粒酶活性和体外增殖 ,可能对肿瘤治疗有潜在的意义。

Objective To study the effect of antisense oligonucleotides of telomerase on the telo merase activity and renal carcinoma cell proliferation. Methods Phosphorothioate antisense oligonucleotides (PS ASON) with sequence TAGGGTTAGACAA,which can recognize the RNA template region of telo merase and PS tandem primer with 13 nucleotides,was incubated with RCZ cell line derived from renal cell carcinoma(RCC).Number of the cells was counted from 1 to 28 days, the inhibiting rate determined by MTT and telomerase activity by PCR ELISA. Results The ASON reduced the telomerase activity ( P <0.001) and the number of cells ( P <0.01) and elevated the inhibiting rate ( P <0.001) as compared with the tandem primer and the empty controls.The efficacy depended on the dose of ASON ( P <0.05) and the sequence selectivity. Conclusions ASON targeted the RNA template inhibited the telomerase activity and RCZ cell proliferation and therefore potentially might be significant in tumor therapy.