不同端粒酶抑制剂对肝癌细胞端粒酶活力的影响  

Effect of different telomerase inhibitors on telomerase activity of hepatocellular carcinoma cells

作　　者：何敏[1] 蒋玉艳[2] 吴华[1] 杨莉[2] 张志勇[2] 李力[3] 

机构地区：[1]广西医科大学医学科学实验中心,广西南宁530021 [2]广西医科大学公共卫生学院劳动卫生教研室,530021 [3]广西壮族自治区肿瘤医院,南宁530021

出　　处：《中国职业医学》2006年第2期79-82,共4页China Occupational Medicine

基　　金：国家自然科学基金项目(30460121);广西自然科学基金项目(桂科青0135008)

摘　　要：目的通过多种端粒酶抑制剂同时作用于人肝癌SMMC-7721细胞,比较各种端粒酶抑制剂抑制癌细胞端粒酶活力的效果。方法利用针对端粒酶RNA的反义寡核苷酸(ASODN)和正义寡核苷酸(NODN),针对端粒酶催化亚基的反义寡核苷酸(HASODN)和正义寡核苷酸(HNODN),以及没食子酰表没食子儿茶素(EGCG),3’-叠氮3’-脱氧胸苷(AZT),全反式维甲酸(ATRA),盐酸阿酶素(ADM)作用于人肝癌SMMC-7721细胞株,实时荧光定量端粒重复序列扩增法(real-timefluorescentquantitativeTRAPassay,FQ-TRAP)检测端粒酶抑制剂作用后SMMC-7721细胞端粒酶活力变化。结果FQ-TRAP法可检测到102个细胞的端粒酶活力,ASODN、NODN、HASODN、HNODN、EGCG、AZT、ADM、ATRA作用于SMMC-7721细胞后,与对照组比较,癌细胞的端粒酶活力均受到抑制(P<0.05),其端粒酶活力的下调率分别为93.30%、43.44%、41.02%、39.26%、58.15%、48.93%、24.44%和53.49%。结论FQ-TRAP法可快速、简便及定量检测人端粒酶活力。ASODN,EGCG抑制SMMC-7721细胞端粒酶活力的效果较明显。Objective To compare the inhibitive effects of telomerase inhibitors on human hepatocelluar cancer SMMC- 7721 cells. Methods SMMC-7721 cells were treated with telomerase inhibitors,antisense oligodeoxynucleotide （ASODN） and sense oligodeoxy-nucleotide （NODN） against hTR gene, antisense oligodeoxynucleotide （hTERT） and sense oligodeoxynucleotide（HNODN） against hTERT gene, EGCG,AZT,ATRA,and ADM. Telomerase activities of SMMC-7721 cells were detected by a novel real-time fluorescent quantitative TRAP （FQ-TRAP） assay. Results The telomerase activities of 100 cells in cell extract were successfully detected by FQ-TRAP assay. Compared with the control, telomerase activities of SMMC-7721 cells were inhibited by vavious telomerase inhibitors, ASODN, NODN, HASODN, HNODN, EGCG, AZT, ADM, and ATRA. The rates of down-regulation of telomerase activity were 93.30% ,43.44% ,41.02% ,39. 26% ,58.15% ,48.93% ,24. 44% and 53.49% , respectively. Conclusion It was suggested that FQ-TRAP could be a rapid and convenient method of determination for telomerase activity. ASODN and EGCG could obviously inhibit telomerase activity of SMMC-7721 cells.

关 键 词：端粒酶活力 端粒酶抑制剂 实时荧光定量端粒重复序列扩增法 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]