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This Article Full Text Full Text (PDF) 081828.Supplemental Data All Versions of this Article: clinchem.2006.081828v1 53/7/1206    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (5) Citing Articles via Google Scholar Google Scholar Articles by Xi, L. Articles by Godfrey, T. E. Search for Related Content PubMed PubMed Citation Articles by Xi, L. Articles by Godfrey, T. E. Related Collections Molecular Diagnostics and Genetics Cancer Diagnostics (since 2002)

Optimal Markers for Real-Time Quantitative Reverse Transcription PCR Detection of Circulating Tumor Cells from Melanoma, Breast, Colon, Esophageal, Head and Neck, and Lung Cancers

¹ Mount Sinai School of Medicine, New York, NY.
² Department of Surgery, University of Pittsburgh, Pittsburgh, PA.
³ Heart, Lung, and Esophageal Surgery Institute, Pittsburgh, PA.

^aAddress correspondence to this author at: Mount Sinai School of Medicine, Box 1079, 1 Gustave L. Levy Pl., New York, NY 10029. Fax 212-241-5432; e-mail: tony.godfrey{at}mssm.edu.

Background: The detection of circulating tumor cells (CTCs) may prove useful for screening, prognostication, and monitoring of response to therapy. However, given the large background of circulating cells, it is probably necessary to detect 1 cancer cell in >10⁶ leukocytes. Although reverse transcription (RT)-PCR is potentially sensitive and specific enough to achieve this goal, success will require the use of appropriate mRNA markers. The goal of this study was to identify optimal marker combinations for detection of CTCs.

Methods: An extensive literature and internet database survey was conducted to identify potential markers. We then used real-time quantitative RT-PCR to test for expression of selected potential markers in tissue samples from primary tumors of breast, colon, esophagus, head and neck, lung, and melanoma and normal blood samples. Markers with high expression in tumors and a median 1000-fold lower expression in normal blood were considered potentially useful for CTC detection and were tested further in an expanded sample set.

Results: A total of 52 potential markers were screened, and 3–8 potentially useful markers were identified for each tumor type. The mRNAs for all but 2 markers were found in normal blood. Marker combinations were identified for each tumor type that had a minimum 1000-fold higher expression in tumors than in normal blood.

Conclusions: Several mRNA markers may be useful for RT-PCR–based detection of CTCs from each of 6 cancer types. Quantification of these mRNAs is essential to distinguish normal expression in blood from that due to the presence of CTCs. Few markers provide adequate sensitivity individually, but combinations of markers may produce good sensitivity for detection of the presence of these 6 neoplasms.

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