荧光定量PCR法检测肿瘤细胞端粒酶活性的变化  

Using fluorescence quantitative PCR to detect telomerase activity in human hepatocelluar carcinoma cells

作　　者：蒋玉艳[1] 何敏[2] 杨莉[2] 李开鹏 

机构地区：[1]广西壮族自治区疾病预防控制中心,南宁530021 [2]广西医科大学公共卫生学院,南宁530021 [3]广西南宁市农业局,南宁530028

出　　处：《现代生物医学进展》2006年第7期12-16,共5页Progress in Modern Biomedicine

基　　金：广西科技厅资助项目(基金编号;桂科青0135008)

摘　　要：目的:利用荧光定量PCR法检测端粒酶抑制剂作用于人肝癌细胞SMMC-7721后端粒酶活性的变化,探讨其抑制端粒酶活性的可能机制,为端粒酶抑制剂的临床应用提供理论依据。方法:利用荧光染料SYBR-Green I建立一种新的端粒酶活性检测方法:FQ-TRAP法。利用FQ-TRAP法检测端粒酶抑制剂作用后肿瘤细胞端粒酶活性变化。结果:端粒酶抑制剂作用后,肝癌细胞端粒酶活性都有变化,其中以ASODN,EGCG,AZT抑制效果较明显。结论:端粒酶FQ-TRAP法是一种特异性、灵敏度、重复性都较好,可快速、简便及定量检测人端粒酶活性的方法,端粒酶抑制剂作用后癌细胞端粒酶活性的变化,为端粒酶抑制剂的临床应用提供理论依据。Objective： To investigate the mechanisms of telomerase inhibitors in the inhibition of telomerase activity and offer theoretical foundation for its clinical application. Methods： Through combining real - time quantitative PCR with TRAP assay and SYBR - Green I, a novel method （FQ- TRAP assay） quantitative detecting telomerase activity was established in the present study. The telomerase activity of carcinoma cells influenced by telomerase inhibitors was detected by FQ - TRAP assay. Results： The telomerase activity of 100 cells in cell extract was detected by a novel real - time quantitativetelomerase detection method （ FQ - TRAP）. The FQ - TRAP is a sensitive, special and repetitive method of telomerase activity assay. Telomerase activity of carcinoma cells influenced by vavious telomerase inhibitors was detected by FQ - TRAP assay. The down - regulation of telomerase activity of cells treated with vavions telomerase inhibitors was observed in every group, obviously in ASODN, EGCG, AZT groups in SMMC- 7721 cells. Conclusion： FQ- TRAP is a rapid, sensitive, specifical and reliable method to quantify telonlerase activity.

关 键 词：定量PGR 肝癌细胞 端粒酶抑制剂 端粒酶活性 

分 类 号：R446.6[医药卫生—诊断学]