Enumeration of Circulating Tumor Cells in the Blood of Breast Cancer Patients After Filtration Enrichment: Correlation with Disease Stage
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The biological and clinical significance of circulating tumor cells (CTC) in the peripheral blood of breast cancer patients is not known. To study this question, we used a direct visualization assay to correlate the number of CTC with disease stage and progression. The CTC were enriched from the nucleated cell fraction by filtration and enumerated visually following immunostaining with anti-cytokeratin 8 (CK8) antibody CAM 5.2. In mixing experiments, we achieved a limit of detection of 5 MCF7 cells per 5 ml of blood or 5 × 107 peripheral blood leukocytes (PBL). We did not detect CTC in any control subjects (0/20). In 131 breast cancer patients, we found a higher incidence of CTC in patients with distant metastatic 36/51 (71%) than those with node-positive 17/36 (47%)(p= 0.026), or node-negative 17/44 (39%)(p= 0.001) disease. The distribution of the highest numbers of CTC observed in individual patients by repeated sampling over time ranged from 1 to 700 per 5 ml of blood with a trend toward higher numbers in those with distant metastases. In comparison with previous studies of equal specificity, based on a similar absence of CTC in controls, we report a higher incidence of CTC in node-negative and node-positive patients, suggesting a more frequent detection of CTC by our approach. This higher incidence was achieved, in part, by repeated sampling of our study population over time. Our results support the concept that CTC can be detected and enumerated in peripheral blood and that this minimally invasive assay merits further evaluation as a potential prognostic indicator and marker of disease progression.
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- 2.Schoenfeld A, Kruger KH, Gomm J, Sinnett HD, Gazet JC, Sacks N, Bender HG, Luqmani Y, Coombes RC: The detection of micrometastases in the peripheral blood and bone marrow of patients with breast cancer using immunohistochemistry and reverse transcriptase polymerase chain reaction for keratin 19. Eur J Cancer 33: 854–861, 1997CrossRefPubMedGoogle Scholar
- 9.Smith BM, Slade MJ, English J, Graham H, Lüchtenborg M, Sinnett HD, Cross NCP, Coombes C: Response of circulating tumor cells to systemic therapy in patients with metastatic breast cancer: comparison of quantitative polymerase chain reaction and immunocytochemical techniques. J Clin Oncol 18: 1432–1439, 2000PubMedGoogle Scholar
- 14.Witzig TE, Bossy B, Kimlinger T, Roche PC, Ingle JN, Grant C, Donohue J, Suman VJ, Harrington D, Torre–Bueno J, Bauer KD: Detection of circulating cytokeratinpositive cells in the blood of breast cancer patients using immunomagnetic enrichment and digital microscopy. Clin Cancer Res 8: 1085–1091, 2002PubMedGoogle Scholar
- 19.Braun S, Pantel K, Muller P, Janni W, Hepp F, Kentenich CRM, Gastroph S, Wischnik A, Dimpfl T, Kindermann G, Riethmuller G, Schlimok G: Cytokeratinpositive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. New Engl J Med 342: 525–533, 2000CrossRefPubMedGoogle Scholar
- 20.Braun S, Hepp F, Kentenich CRM, Janni W, Pantel K, Riethmüller G, Willgeroth F, Sommer HL: Monoclonal antibody therapy with edrecolomab in breast cancer patients: monitoring of elimination of disseminated cytokeratin–positive tumor cells in bone marrow. Clin Cancer Res 5: 3999–4004, 1999 HJ Kahn et al. 246Google Scholar
- 21.Ménard S, Squicciarini P, Luini A, Sacchini V, Rovini D, Tagliabue E, Veronesi P, Salvadori B, Veronesi U, Colnaghi MI: Immunodetection of bone marrow micrometastases in breast carcinoma patients and its correlation with primary tumour prognostic features. Brit J Cancer 69: 1126–1129, 1994PubMedGoogle Scholar