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Received on November 5, 2004
Accepted on March 17, 2005
Cancer Diagnostics |
1 Department of Molecular Oncology, UCLA School of Medicine, Los Angeles, CA
2 Department of Biomathematics, UCLA School of Medicine, Los Angeles, CA
3 Division of Surgical Oncology, John Wayne Cancer Institute at Saint John's Health Center, Santa Monica, CA
4 Cancer Institute Medical Group, Santa Monica, CA
5 Department of Molecular Oncology
* To whom correspondence should be addressed. E-mail: hoon{at}jwci.org.
Background: Detection of melanoma cells in circulation may be important in assessing tumor progression. The objective of this study was to develop a specific, reliable, multimarker quantitative real-time reverse transcription-PCR (qRT) assay for detecting melanoma cells in patients' blood.
Methods: We developed qRT assays for the mRNA of four melanoma-associated markers: MART-1, GalNAc-T, PAX-3, and MAGE-A3. In optimization studies, we tested 17 melanoma cell lines and 49 peripheral blood leukocyte (PBL) samples from volunteers. We performed RNA and melanoma cell dilution studies to assess the detection limits and imprecision of the assays. We measured the mRNAs in blood specimens from 94 melanoma patients [American Joint Committee on Cancer (AJCC) stage I, n = 20; II, n = 20; III, n = 32; IV, n = 22].
Results: All markers were frequently detected in melanoma cell lines, whereas none of the markers was detected in PBLs from volunteers. The qRT assay could detect 1 melanoma cell in 107 PBLs in the melanoma cell-dilution studies. Markers were detected in 15%, 30%, 75%, and 86% of melanoma patients with AJCC stage I, II, III, and IV disease, respectively. The number of positive markers and AJCC stage were significantly correlated (Spearman correlation coefficient = 0.58; P <0.0001).
Conclusions: Multimarker qRT can detect circulating melanoma cells in blood. Measurement of the studied molecular markers in blood may be useful in detection of metastasis and monitoring treatment response of melanoma patients.
The following articles in journals at HighWire Press have cited this article:
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T. Nakagawa, S. R. Martinez, Y. Goto, K. Koyanagi, M. Kitago, T. Shingai, D. A. Elashoff, X. Ye, F. R. Singer, A. E. Giuliano, et al. Detection of Circulating Tumor Cells in Early-Stage Breast Cancer Metastasis to Axillary Lymph Nodes Clin. Cancer Res., July 15, 2007; 13(14): 4105 - 4110. [Abstract] [Full Text] [PDF]
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S. Mocellin, D. Hoon, A. Ambrosi, D. Nitti, and C. R. Rossi The Prognostic Value of Circulating Tumor Cells in Patients with Melanoma: A Systematic Review and Meta-analysis Clin. Cancer Res., August 1, 2006; 12(15): 4605 - 4613. [Abstract] [Full Text] [PDF]
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K. Koyanagi, S. J. O'Day, R. Gonzalez, K. Lewis, W. A. Robinson, T. T. Amatruda, C. Kuo, H.-J. Wang, R. Milford, D. L. Morton, et al. Microphthalmia Transcription Factor as a Molecular Marker for Circulating Tumor Cell Detection in Blood of Melanoma Patients Clin. Cancer Res., February 15, 2006; 12(4): 1137 - 1143. [Abstract] [Full Text] [PDF]
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K. Koyanagi, S. J. O'Day, R. Gonzalez, K. Lewis, W. A. Robinson, T. T. Amatruda, H.-J. Wang, R. M. Elashoff, H. Takeuchi, N. Umetani, et al. Serial Monitoring of Circulating Melanoma Cells During Neoadjuvant Biochemotherapy for Stage III Melanoma: Outcome Prediction in a Multicenter Trial J. Clin. Oncol., November 1, 2005; 23(31): 8057 - 8064. [Abstract] [Full Text] [PDF]
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