Abstract
The detection of circulating melanoma cells has been the subject of numerous investigations in recent years. We developed a cellular approach to identifying circulating melanoma cells in peripheral blood using immunomagnetic cell sorting. The examination covered 205 blood samples from 155 melanoma patients and 30 samples from healthy persons and nonmelanoma patients. After density gradient centrifugation, the interphase was incubated with the 9.2.27 antibody. Positive cells were labeled with magnetic micro-beads and enriched by immunomagnetic cell sorting. Cells were stained using an alkaline phosphatase—anti-alkaline phosphatase assay and examined by light microscopy. In spiking experiments, melanoma cells seeded at a concentration of one melanoma cell per milliliter of whole blood could be detected reliably. Circulating melanoma cells were not found in 30 controls, nor were 9.2.27-positive cells found in 41 patients with primary malignant melanoma. In patients with regional lymph node metastases and disseminated disease, circulating 9.2.27-positive cells could be detected in 3 of 29 patients (10%) and 13 of 85 patients (15%) examined, respectively. We conclude that immunomagnetic cell sorting is a promising method with high sensitivity and specificity. The method is not suitable for early detection of metastases but is a valuable tool for further investigating the biological characteristics of circulating melanoma cells.
Supported by a grant from the Federal Ministry of Education, Science, Research, and Technology (Fö. 01KS9602) and the Interdisciplinary Clinical Research Center (IKFZ), Tübingen, Germany.
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References
Keilholz U, Willhauck M, Scheibenbogen C, de Vries TJ, Burchill S (1997) Polymerase chain reaction detection of circulating tumour cells. EORTC Melanoma Cooperative Group, Immunotherapy Subgroup. Melanoma Res 7 Suppl 2: S133–141
Curry BJ, Myers K, Hersey P (1998) Polymerase chain reaction detection of melanoma cells in the circulation: relation to clinical stage, surgical treatment, and recurrence from melanoma. J Clin Oncol 16: 1760–1769
Farthmann B, Eberle J, Krasagakis K, Gstottner M, Wang N, Bisson S, Orfanos CE (1998) RT-PCR for tyrosinase-mRNA-positive cells in peripheral blood: evaluation strategy and correlation with known prognostic markers in 123 melanoma patients. J Invest Dermatol 110: 263–267
Ghossein RA, Coit D, Brennan M, Zhang ZF, Wang Y, Bhattacharya S, Houghton A, Rosai J (1998) Prognostic significance of peripheral blood and bone marrow tyrosinase messenger RNA in malignant melanoma. Clin Cancer Res 4: 419–428
Mellado B, Colomer D, Castel T, Munoz M, Carballo E, Galan M, Mascaro JM, Vives Corrons JL, Grau JJ, Estape J (1996) Detection of circulating neoplastic cells by reverse-transcriptase polymerase chain reaction in malignant melanoma: association with clinical stage and prognosis. J Clin Oncol 14: 2091–2097
Curry BJ, Smith MJ, Hersey P (1996) Detection and quantitation of melanoma cells in the circulation of patients. Melanoma Res 6: 45–654
Glaser R, Rass K, Seiter S, Hauschild A, Christophers E, Tilgen W (1997) Detection of circulating melanoma cells by specific amplification of tyrosinase complementary DNA is not a reliable tumor marker in melanoma patients: a clinical two-center study. J Clin Oncol 15: 2818–2825
Reinhold U, Ludtke Handjery HC, Schnautz S, Kreysel HW, Abken H (1997) The analysis of tyrosinase-specific mRNA in blood samples of melanoma patients by RT-PCR is not a useful test for metastatic tumor progression. J Invest Dermatol 108: 166–169
Brossart P, Schmier JW, Kruger S, Willhauck M, Scheibenbogen C, Mohler T, Keilholz U (1995) A polymerase chain reaction-based semiquantitative assessment of malignant melanoma cells in peripheral blood. Cancer Res 55: 4065–4068
Brossart P, Keilholz U, Scheibenbogen C, Mohler T, Willhauck M, Hunstein W (1994) Detection of residual tumor cells in patients with malignant melanoma responding to immunotherapy. J Immunother 15: 38–41
Brossart P, Keilholz U, Willhauck M, Scheibenbogen C, Mohler T, Hunstein W (1993) Hematogenous spread of malignant melanoma cells in different stages of disease. J Invest Dermatol 101: 887–889
Foss AJ, Guile MJ, Occleston NL, Hykin PG, Hungerford JL, Lightman S (1995) The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction. Br J Cancer 72: 155–159
Palmieri G, Strazzullo M, Ascierto PA, Satriano SMR, Daponte A, Castello G (1999) Polymerase chain reaction-based detection of circulating melanoma cells as an effective marker of tumor progression. J Clin Oncol 17: 304–311
Pluschke G, Vanek M, Evans A, Dittmar T, Schmid P, Itin P, Filardo EJ, Reisfeld RA (1996) Molecular cloning of a human melanoma-associated chondroitin sulfate proteoglycan. Proc Natl Acad Sci USA 93: 9710–9715
Heyderman E, McCartney JC (1985) Epithelial membrane antigen and lymphoid cells. Lancet 1: 109–110
Bumol TF, Reisfeld RA (1982) Unique glycoprotein-proteoglycan complex defined by monoclonal antibody on human melanoma cells. Proc Natl Acad Sci USA 79: 1245–1249
Harper JR, Quaranta V, Reisfeld RA (1986) Ammonium chloride interferes with a distinct step in the biosynthesis and cell surface expression of human melanoma-type chondroitin sulfate proteoglycan. J Biol Chem 261: 3600–3606
Oldham RK, Foon KA, Morgan AC, Woodhouse CS, Schroff RW, Abrams PG, Fer M, Schoenberger CS, Farrell M, Kimball E et al. (1984) Monoclonal antibody therapy of malignant melanoma: in vivo localization in cutaneous metastasis after intravenous administration. J Clin Oncol 2: 1235–1244
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Benez, A., Schiebel, U., Fierlbeck, G. (2001). Morphologically Intact Melanoma Cells May Be Detected in Peripheral Blood of Melanoma Patients. In: Reinhold, U., Tilgen, W. (eds) Minimal Residual Disease in Melanoma. Recent Results in Cancer Research, vol 158. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59537-0_11
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DOI: https://doi.org/10.1007/978-3-642-59537-0_11
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