端粒酶反义寡核苷酸对膀胱癌端粒酶活性及细胞增殖的影响

目的 :探讨端粒酶反义核苷酸对膀胱癌的治疗作用。方法 :以端粒酶 RNA模板区为靶点 ,不同剂量的序列为 TAGGGTTAGACAA的硫代磷酸修饰的反义寡核苷酸 ( PS- ASON)作为实验组 ( A、B、C) ,以硫代磷酸修饰 1 3个核苷酸的随机引物作为对照组 ( D) ,仅加入培养基作为空白对照组 ( E) ,与人膀胱癌细胞株 BIU87细胞共同孵育 ,TRAP- ELISA方法检测端粒酶活性变化 ,计数1～ 30 d细胞数 ,计算细胞倍增时间 ,MTT法测定 ASON对细胞的抑制率。结果 :2 5例膀胱癌组织中 ,2 4例 ( 96% )表达端粒酶活性 ,4例癌旁组织无端粒酶活性表达。与随机引物及空白对照组比较 ,实验组的 ASON能明显降低端粒酶活性 ,减少细胞增殖数目 ,增加抑制率及延长倍增时间 ,并显示剂量的依赖性 ( P<0 .0 1 )。结论 :以端粒酶 RNA模板区为靶点的反义寡核苷酸引起膀胱癌 BIU87细胞端粒酶活性下降 ,增殖抑制 ,引起细胞退化 ,对膀胱癌的治疗有潜在的意义。

Effect of Telomerase Antisense Oligonucleotides on the Inhibition of Telomerase Activity and Cell Growth in Bladder Cancer in Vitro

Objective:To investigate the therapeutic effects of the telomerase antisense nucleotides on the BIU87 cell derived from bladder cancer. Methods:The phosphorothioate antisense oligonucleotides(PS ASON) of differential dose with sequence TTAGGGTTAGACAA targeted to telomerase RNA template(as A,B,C experiment group)and PS random primer with 13 nucleotides(as control group) were incubated with the BIU87 cell line.The number of cells was counted from 1 to 30 days,doubling time was calculated and inhibiting rate was determined by MTT method. Results:ASON group reduced telomerase activity and the number of cells,lengthened doubling time,and elevated the inhibiting rate comtrasted with random primer control and empty control.The efficacy depended on the dose of ASON ( P <0.05) and it was of sequence selective specifity. Conclusion:The ASON targeted hTR has an inhibition effect on the telomerase activity and growth of BIU87 cell, induce cells degraduation might have potential significance in tumor therapy.