Switch to
Predicate | Object |
---|---|
rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
|
pubmed:dateCreated |
1997-10-23
|
pubmed:abstractText |
Conflicting results have been reported on the use of cytokeratin-19 (CK-19) in the detection of tumor cells in the peripheral blood of patients with solid tumors. We investigated the expression of CK-19 in lung cancer cell lines and in human lung tumor samples using a nested reverse transcriptase (RT)-PCR to determine the sensitivity and specificity of this method. In addition, blood samples of lung cancer patients and healthy controls were analyzed for the presence of CK-19 transcripts. Amplification products were visualized by ethidium bromide staining and radioactive hybridization with a CK-19-specific probe. Application of a previously described nested RT-PCR for the detection of CK-19 resulted in amplification of the processed pseudogene. Therefore, a more stringent RT-PCR was developed by increasing the annealing temperature. RT-PCR amplification products for CK-19 were detected in 38 of 41 lung cancer cell lines. The three negative cell lines were all variant small-cell lung cancer cell lines. Concordant results were observed between CK-19 detection by immunohistochemistry and by RT-PCR. In serial RNA dilution experiments, CK-19 transcripts could be detected in 18 to 80 pg of total cellular RNA in three cell lines and in 60 ng total RNA in one cell line. The nested RT-PCR had the sensitivity of detecting 50 tumor cells in 10(6) peripheral blood mononuclear cells (PBMNC), and CK-19 transcripts were randomly detected in normal PBMNC. This study shows the necessity in processing parallel samples without reverse transcriptase enzyme to avoid amplification of pseudogenes. A serious problem in the detection of tissue-specific transcripts in PBMNC is the detection of illegitimate transcription levels. In conclusion, although CK-19 may be a useful marker for the detection of lung cancer cells, its application for the detection of circulating tumor cells is not recommended.
|
pubmed:commentsCorrections | |
pubmed:language |
eng
|
pubmed:journal | |
pubmed:citationSubset |
IM
|
pubmed:chemical | |
pubmed:status |
MEDLINE
|
pubmed:month |
Sep
|
pubmed:issn |
0023-6837
|
pubmed:author | |
pubmed:issnType |
Print
|
pubmed:volume |
77
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
213-20
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:9314945-Blood Cells,
pubmed-meshheading:9314945-Humans,
pubmed-meshheading:9314945-Immunohistochemistry,
pubmed-meshheading:9314945-Keratins,
pubmed-meshheading:9314945-Lung,
pubmed-meshheading:9314945-Lung Neoplasms,
pubmed-meshheading:9314945-Polymerase Chain Reaction,
pubmed-meshheading:9314945-RNA, Messenger,
pubmed-meshheading:9314945-Reference Values,
pubmed-meshheading:9314945-Sensitivity and Specificity,
pubmed-meshheading:9314945-Transcription, Genetic,
pubmed-meshheading:9314945-Tumor Cells, Cultured
|
pubmed:year |
1997
|
pubmed:articleTitle |
Detection of cytokeratin-19 transcripts by reverse transcriptase-polymerase chain reaction in lung cancer cell lines and blood of lung cancer patients.
|
pubmed:affiliation |
Department of Medical Oncology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|