| 蒋玉艳1 何 敏2 杨 莉2 李开鹏3.荧光定量PCR 法检测肿瘤细胞端粒酶活性的变化[J].,2006,6(7):12-16 |
| 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 activityin human hepatocelluar carcinoma cells |
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| DOI： |
| 中文关键词: 定量PCR 肝癌细胞 端粒酶抑制剂 端粒酶活性 |
| 英文关键词: The real - time quantitative PCR Hepatocelluar cancer cell Telomer ase inhibitors Telomerase activity |
| 基金项目:广西科技厅资助项目( 基金编号, 桂科青0135008) |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 carc-i
noma 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 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. |
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