端粒酶基因hTRT反义寡核苷酸对其酶活性和肿瘤细胞增殖的影响  

THE INFLUENCE OF TELOMERASE hTRT GENE ANTISENSE OLIGONUCLEOTIDE ON ITS ENZYMIC ACTIVITY AND PROLIFERATION OF TUMOR CELL

作　　者：李贵新[1] 赵常在[1] 王会东[1] 张永庆[1] 石毅然[1] 张建华[1] 

机构地区：[1]山东大学临床医学院山东省立医院肿瘤研究治疗中心,济南250021

出　　处：《青岛大学医学院学报》2002年第1期10-13,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省科学技术攻关计划资助项目 (98115 45 0 3B)

摘　　要：①目的　探讨端粒酶基因hTRT反义寡核苷酸对肿瘤细胞增殖和端粒酶活性的影响。②方法　采用MTT法检测反义寡核苷酸对肿瘤细胞HL 6 0增殖的影响 ;采用TRAP PCR法检测反义寡核苷酸处理肿瘤细胞后端粒酶活性的变化 ;采用Giemsa染色观察反义寡核苷酸处理后肿瘤细胞形态学变化 ;采用DNA凝胶电泳和三苯胺法检测反义寡核苷酸处理后肿瘤细胞DNA片段化。③结果　端粒酶基因hTRT反义寡核苷酸作用 4 8h对肿瘤细胞增殖出现抑制作用。 5 μmol/L反义寡核苷酸处理 2 4h后 ,细胞端粒酶活性下降 ,并且随作用时间的延长 ,活性不断降低 ,96h下降至对照组的 4 4 .4 % .用 5 μmol/L反义寡核苷酸处理 96h的HL 6 0细胞出现凋亡的形态学特征 ,处理 2 4 ,4 8,72 ,96h后凋亡指数为 0 .0 5 3,0 .12 7,0 .2 5 7,0 .315 .用 5 μmol/L反义寡核苷酸处理HL 6 0细胞 96h后 ,电泳出现DNA梯状条带。作用 2 4 ,4 8,72 ,96h细胞DNA片段化率分别为 17.98% ,2 9.34% ,35 .4 3% ,4 1.2 4 % .④结论　端粒酶hTRT基因反义寡核苷酸能抑制肿瘤细胞的增殖和端粒酶活性 ,并能诱导肿瘤细胞HL 6Objective\ To study the effect of human telomerase reverse transcriptase antisense oligonucleotide(hTRT ASO)on its enzymic activity and proliferation of tumor cells. \ Methods\ MTT assay was used to detect the effect of hTRT ASO on proliferation of tumor cell line HL 60. Telomeres repeat amplification protocol(TRAP PCR) was used to study the changes of telomerase activity of tumor cells treated by hTRT ASO. Giemsa stain and light microscope were used to observe the morphological changes of tumor cells after treatment by hTRT ASO.DNA gel electrophoresis and diphenylamine assay were used to test DNA fragmentation of HL 60 cells treated by hTRT ASO. \ Results\ The proliferation of HL 60 cells was inhibited after treatment by hTRT ASO for 48 hours. The telomerase activity decreased after treatment by hTRT ASO(5μmol/L) for 24 hours. The telomerase activity decreased constantly with prolongation of time. The telomerase activity of tumor cell treated for 96 hours was 44.4% of the control. HL 60 cells treated by hTRT ASO (5μmol/L) for 96 hours appeared standard apoptotic morphological characters. Apoptotic indexes of HL 60 treated for 24, 48, 72 and 96 hours were 0.053, 0.127, 0.257 and 0.315, respectively. DNA ladder of HL 60 cells was detected by gel electrophoresis after treatment by hTRT ASO (5μmol/L )for 96 hours. DNA fragmentation ratio of HL 60 cells treated by hTRT ASO for 24, 48, 72 and 96 hours was 17.98%, 29.34%, 35.43% and 41.24% respectively. Conclusion\ hTRT ASO can inhibit the proliferation of tumor cells and the activity of telomerase and induce apoptosis of tumor cells.

关 键 词：端粒酶类基因 寡核苷酸类 肿瘤细胞 

分 类 号：R730.2[医药卫生—肿瘤]