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学科主题: 基础医学
题名:
Detection of telomerase activity in cultured cells and tumor tissue of lung carcinoma by modified telomeric repeat amplification protocol
作者: Liu, Yan1,2; Wu, Bing-quan1; Zhong, Hao-hao1; Xu, Mei-lin2; Fang, Wei-gang1
关键词: cancer ; real-time polymerase chain reaction ; SYBR green ; telomerase ; telomeric repeat amplification protocol
刊名: PATHOLOGY INTERNATIONAL
发表日期: 2010-05-01
DOI: 10.1111/j.1440-1827.2010.02529.x
卷: 60, 期:5, 页:386-394
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Pathology
研究领域[WOS]: Pathology
关键词[WOS]: QUANTITATIVE-ANALYSIS ; BRONCHIAL LAVAGE ; CANCER ; ASSAY ; QUANTIFICATION ; EXPRESSION ; MARKER ; HTERT
英文摘要:

Telomerase activity is found in various cell types including stem cells, neoplastic cells, and immortalized cells, suggesting a close association with their proliferation capacity. The telomeric repeat amplification protocol (TRAP) has been traditionally used to detect semi-quantitatively the telomerase activity by polyacrylamide gel electrophoresis (PAGE), which is difficult to apply for large scale analysis because of laborious post-PCR manipulation and potential carryover contamination. In the present study, a specific reverse primer was designed and the TRAP protocol was adapted to either PAGE or real-time PCR assay. Using cultured cell lines, the real-time TRAP showed a dramatic improvement in the reliability and accuracy of quantitation of telomerase activity and was able to discriminate the A549 cells from hundreds-fold human embryonic lung cells. Using clinical samples of 60 lung cancers and 8 inflammatory lesions, the real-time TRAP was also superior in quantitation, high-throughput capability and standardization. Our modified real-time TRAP should be applicable for the detection of telomerase activity for the initial screening and progression monitoring of lung cancer patients. Our approach is particularly useful when only limited clinical specimen is available, such as fine needle aspiration or other cytological specimens that may contain only a small number of tumor cells.

语种: 英语
WOS记录号: WOS:000277205100007
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/58338
Appears in Collections:基础医学院_病理学系_期刊论文

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作者单位: 1.Peking Univ, Hlth Sci Ctr, Dept Pathol, Beijing 100191, Peoples R China
2.Tianjin Chest Hosp, Tianjin, Peoples R China

Recommended Citation:
Liu, Yan,Wu, Bing-quan,Zhong, Hao-hao,et al. Detection of telomerase activity in cultured cells and tumor tissue of lung carcinoma by modified telomeric repeat amplification protocol[J]. PATHOLOGY INTERNATIONAL,2010,60(5):386-394.
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