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1 Immunicon Corporation, Huntingdon Valley, Pennsylvania; 2 Fels Institute for Cancer Research, Temple University School of Medicine; 3 Department of Medical Oncology, Fox Chase Cancer Center; 4 Department of Urology, Thomas Jefferson University Hospital, Philadelphia, Pennsylvania; and 5 Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan
Requests for reprints: S. Mark O'Hara, Immunicon Corporation, Research and Development, 3401 Masons Mill Road, Suite 100, Huntingdon Valley, PA 19006. Phone: 215-830-0777; Fax: 215-830-0751; E-mail: smohara{at}immunicon.com.
Metastases from primary tumors are responsible for most cancer deaths. It has been shown that circulating tumor cells (CTCs) can be detected in the peripheral blood of patients with a variety of metastatic cancers and that the presence of these cells is associated with poor clinical outcomes. Characterization of CTCs in metastatic cancer patients could provide additional information to augment management of the disease. Here, we describe a novel approach for the identification of molecular markers to detect and characterize CTCs in peripheral blood. Using an integrated platform to immunomagnetically isolate and immunofluorescently detect CTCs, we obtained blood containing 
100 CTCs from one metastatic colorectal, one metastatic prostate, and one metastatic breast cancer patient. Using the RNA extracted from the CTC-enriched portion of the sample and comparing it with the RNA extracted from the corresponding CTC-depleted portion, for the first time, global gene expression profiles from CTCs were generated and a list of cancer-specific, CTC-specific genes was obtained. Subsequently, samples immunomagnetically enriched for CTCs from 74 metastatic cancer patients and 50 normal donors were used to confirm by quantitative real-time reverse transcription-PCR CTC-specific expression of selected genes and to show that gene expression profiles for CTCs may be used to distinguish normal donors from advanced cancer patients as well as to differentiate among the three different metastatic cancers. Genes such as AGR2, S100A14, S100A16, FABP1, and others were found useful for detection of CTCs in peripheral blood of advanced cancer patients.
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