Document Detail


Detection of telomerase activity in cultured cells and tumor tissue of lung carcinoma by modified telomeric repeat amplification protocol.
MedLine Citation:
PMID:  20518889     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Yan Liu; Bing-quan Wu; Hao-hao Zhong; Mei-lin Xu; Wei-gang Fang
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Pathology international     Volume:  60     ISSN:  1440-1827     ISO Abbreviation:  Pathol. Int.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-06-03     Completed Date:  2010-09-02     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9431380     Medline TA:  Pathol Int     Country:  Australia    
Other Details:
Languages:  eng     Pagination:  386-94     Citation Subset:  IM    
Affiliation:
Department of Pathology, Peking University Health Science Center, Beijing, China.
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MeSH Terms
Descriptor/Qualifier:
Adult
Aged
Aged, 80 and over
Carcinoma, Non-Small-Cell Lung / enzymology*
Cell Line, Tumor
Electrophoresis, Gel, Two-Dimensional
Female
High-Throughput Screening Assays
Humans
Lung Neoplasms / enzymology*,  genetics
Male
Middle Aged
Nucleic Acid Amplification Techniques / methods*
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Telomerase / metabolism*
Telomere / genetics*
Terminal Repeat Sequences / genetics
Chemical
Reg. No./Substance:
EC 2.7.7.49/Telomerase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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