Application of quantitative RT-PCR using 'TaqMan' technology to evaluate the expression of CK 18 mRNA in various cell lines

E. Tokunaga, Yoshihiko Maehara, Eiji Oki, T. Koga, Y. Kakeji, K. Sugimachi

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Reverse transcriptase polymerase chain reaction (RT-PCR) is often used for sensitive detection of micrometastasis in peripheral blood, lymph nodes and bone marrow. While the utility of this method has been documented, it also has limitations in the detection of micrometastasis. The mRNA of target genes can be detected in healthy donors or in samples used for negative control, therefore the non-quantitativeness of conventional RT-PCR has been called into question. We analyzed the expression level of cytokeratin (CK) 18 mRNA in established esophageal and gastrointestinal carcinoma cell lines and non-epithelial cells, using quantitative RT-PCR, based on real time 'TaqMan TM' technology. CK 18 mRNA is more highly expressed in carcinoma cells than in non-epithelial cells. However, the expression level in non-epithelial cells was easily detected using conventional RT-PCR and agarose gel electrophoresis. In an analysis of CK 18 mRNA expression in peripheral venous blood in 13 healthy volunteers, we found that CK 18 mRNA was much less expressed than in cancer cell lines. However, the expression in all samples was at a level which was also detected using conventional RT-PCR. It would thus seem that not only qualitative, but also quantitative analysis, of the target mRNA is important to detect micrometastasis. Quantitative RT-PCR methods will make comparisons of the possible differences in expression levels of the target gene. For clinical applications, much further study is needed.

Original languageEnglish
Pages (from-to)375-381
Number of pages7
JournalJournal of Experimental and Clinical Cancer Research
Volume19
Issue number3
Publication statusPublished - Nov 25 2000

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Keratin-18
Reverse Transcriptase Polymerase Chain Reaction
Technology
Cell Line
Neoplasm Micrometastasis
Messenger RNA
Carcinoma
Agar Gel Electrophoresis
Genes
Healthy Volunteers
Lymph Nodes
Bone Marrow
Tissue Donors
Neoplasms

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Application of quantitative RT-PCR using 'TaqMan' technology to evaluate the expression of CK 18 mRNA in various cell lines. / Tokunaga, E.; Maehara, Yoshihiko; Oki, Eiji; Koga, T.; Kakeji, Y.; Sugimachi, K.

In: Journal of Experimental and Clinical Cancer Research, Vol. 19, No. 3, 25.11.2000, p. 375-381.

Research output: Contribution to journalArticle

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