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Melanoma Research:
doi: 10.1097/CMR.0000000000000025
ORIGINAL ARTICLES: Translational research

Prevalence and heterogeneity of circulating tumour cells in metastatic cutaneous melanoma

Khoja, Leilaa,b,c; Shenjere, Patrickb; Hodgson, Clarea; Hodgetts, Jackieb; Clack, Glend; Hughes, Andrewd; Lorigan, Paulc; Dive, Carolinea,c

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Abstract

We previously demonstrated that circulating tumour cells (CTCs) are detectable by the MelCAM and high molecular weight melanoma-associated antigen (HMW-MAA)-dependent CellSearch platform. However, CTCs which do not express these capture and detection markers are not detectable by CellSearch. Consequently, we explored the use of isolation by size of epithelial tumour cells (ISET), a marker independent, filtration-based device to determine the prevalence and heterogeneity of CTCs in metastatic cutaneous melanoma patients. Ninety patients were prospectively recruited and blood samples taken before treatment. Patients’ blood was filtered using the ISET platform. CTCs were enumerated using dual immunohistochemistry with positive selection by S100 expression and exclusion of leucocytes and endothelial cells expressing CD45 or CD144, respectively. A panel of markers (Melan-A, MITF, MelCAM, high molecular melanoma-associated antigen, CD271 and MAGEC) was also examined. Fifty-one patients (57%) had CTCs (range 1–44 CTCs/4 ml blood) and 12 patients also had circulating tumour microemboli. Seven patients had S100− CTCs, 11 patients’ CTCs were S100+ and 33 patients had S100+ and S100− CTCs. Substantial intrapatient and interpatient heterogeneity was observed for all other melanoma-associated markers. CTCs in metastatic cutaneous melanoma are detectable using the flexible marker-independent ISET platform. CTCs display significant marker expression heterogeneity implying that marker-dependent platforms would not detect all CTCs and multimarker assays are now required to reveal the biological significance of this CTC heterogeneity.

© 2014 Wolters Kluwer Health | Lippincott Williams & Wilkins

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