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Circulating Tumor Cell Analysis in Patients with Progressive Castration-Resistant Prostate Cancer

Authors' Affiliations: ¹ Genitourinary Oncology Service, Department of Medicine, ² Molecular Cytogenetics Core Laboratory, Departments of ³ Pathology and ⁴ Clinical Laboratories, ⁵ Leukemia Service, Department of Medicine, ⁶ Immunicon Corporation, Huntingdon Valley, Pennsylvania, ⁷ Urology Service, Department of Surgery, and ⁸ Department of Epidemiology and Biostatistics, Memorial Sloan-Kettering Cancer Center, and ⁹ Department of Medicine, Joan and Sanford E. Weill College of Medicine of Cornell University, New York, New York

Requests for reprints: Howard I. Scher, Genitourinary Oncology Service, Department of Medicine, Sidney Kimmel Center for Prostate and Urologic Cancers, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021. Phone: 646-422-4323; Fax: 212-988-0851; E-mail: Scherh{at}mskcc.org or Martin Fleisher, Department of Laboratory Medicine, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021. E-mail: Fleishem{at}mskcc.org.

Purpose: To better direct targeted therapies to the patients with tumors that express the target, there is an urgent need for blood-based assays that provide expression information on a consistent basis in real time with minimal patient discomfort. We aimed to use immunomagnetic-capture technology to isolate and analyze circulating tumor cells (CTC) from small volumes of peripheral blood of patients with advanced prostate cancer.

Experimental Design: Blood was collected from 63 patients with metastatic prostate cancer. CTCs were isolated by the Cell Search system, which uses antibodies to epithelial cell adhesion marker and immunomagnetic capture. CTCs were defined as nucleated cells positive for cytokeratins and negative for CD45. Captured cells were analyzed by immunofluorescence, Papanicolau staining, and fluorescence in situ hybridization.

Results: Most patients (65%) had 5 or more CTCs per 7.5 mL blood sample. Cell counts were consistent between laboratories (c = 0.99) and did not change significantly over 72 or 96 h of storage before processing (c = 0.99). Their identity as prostate cancer cells was confirmed by conventional cytologic analysis. Molecular profiling, including analysis of epidermal growth factor receptor (EGFR) expression, chromosome ploidy, and androgen receptor (AR) gene amplification, was possible for all prostate cancer patients with 5 CTCs.

Conclusions: The analysis of cancer-related alterations at the DNA and protein level from CTCs is feasible in a hospital-based clinical laboratory. The alterations observed in EGFR and AR suggest that the methodology may have a role in clinical decision making.

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