端粒酶反义寡核苷酸诱导肝癌细胞凋亡的实验  

Experimental of Apoptosis of HCC Induced by Telomerase-antisense Oligo Nucleotide

作　　者：邓志华[1] 杨长青[1] 刘燕[1] 刘金龙[1] 杨强[1] 赵婕[1] 韩子岩[1] 曹燕[1] 

机构地区：[1]山西医科大学第二医院消化科,太原030002

出　　处：《肿瘤防治研究》2009年第3期171-174,共4页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金资助项目(30672405);山西省青年基金资助项目(20001028)

摘　　要：目的研究端粒酶反义寡核苷酸对肝癌细胞株的凋亡诱导作用及其机制。方法以人肝癌细胞株SMMC-7721、正常肝细胞株L-02为研究对象,采用TRAP-ELISA法对肿瘤细胞端粒酶活性进行定性定量分析;细胞形态学、TUNEL染色、DNA琼脂糖凝胶电泳观察端粒酶反义寡核苷酸对肝癌细胞的凋亡诱导作用,用流式细胞仪对细胞周期进行时相分析,Western杂交对细胞周期蛋白进行研究。结果一定浓度的端粒酶反义寡核苷酸可抑制肝癌细胞端粒酶活性,肝癌细胞端粒酶活性被抑制后传代23～26次出现细胞凋亡。细胞凋亡与细胞周期蛋白有关。结论端粒酶反义寡核苷酸可抑制肝癌细胞端粒酶活性并诱导其凋亡。Objective To study the effect of telomerase-antisense oligonucleotide on the induction of apoptosis of hepatoeellular carcinoma cell line and its mechanism. Methods We used HCC cell line SMMC- 7721 as a target and cell line L-02 as the normal. Using the TRAP-ELISA we quantitively and qualitatively studied telomerase activity. We detected the apoptosis of HCC cell induced by telomerase-antisense oligonucleotide through TUNEL stain, morphology and DNA electrophoresis. The phase analysis was performed by flow cytometry. We also used Western blot to study the expressing of cell cycle proteins. Results Telomerase-antisense oligonucliotide at proper concentration can obviously inhibit the telomerase activity of hepatocellularcarcinoma cell line and induce apoptosis after the cell were passageed for 23 to 26 times. The apoptosis is related with the expressing of cell cycle proteins. Conclusion Telomerase-antisense oligonucleotide can inhibit the telomerase activity of hepatocellularcarcinoma cell line and induce its apoptosis.

关 键 词：端粒酶 肝癌 反义寡核苷酸 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]