荧光定量PCR 法检测肿瘤细胞端粒酶活性的变化

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。

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

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 realtime 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 vavious 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 telomerase activity.