Application of a Filtration- and Isolation-by-Size Technique for the Detection of Circulating Tumor Cells in Cutaneous Melanoma Vincenzo De Giorgi, Pamela.

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Presentation transcript:

Application of a Filtration- and Isolation-by-Size Technique for the Detection of Circulating Tumor Cells in Cutaneous Melanoma Vincenzo De Giorgi, Pamela Pinzani, Francesca Salvianti, John Panelos, Milena Paglierani, Agata Janowska, Marta Grazzini, Janine Wechsler, Claudio Orlando, Marco Santucci, Torello Lotti, Mario Pazzagli, Daniela Massi Journal of Investigative Dermatology Volume 130, Issue 10, Pages 2440-2447 (October 2010) DOI: 10.1038/jid.2010.141 Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Morphological and immunohistochemical identification of circulating melanoma cells. (a) ISET-positive samples display isolated circulating melanoma cells. Neoplastic cells fulfill criteria for circulating tumor cells, including: (i) cell size ≥16μm, (ii) nucleo-cytoplasmic ratio ≥50%, (iii) irregular nuclear shape, (iv) hyperchromatic nucleus, and (v) basophilic cytoplasm (upper row, original magnification × 63, hematoxylin and eosin stain). Immunohistochemical analyses with anti-S100 protein, MART-1, HMB-45 antibodies identify circulating melanoma cells as weak to moderately stained cells. CD45 is negative (lower row, original magnification × 63). (b) Melanoma cell lines (SK-MEL-28) immunostained in artificial samples with S100 protein, MART-1, HMB-45, and CD45. Note that the intensity of the immunostaining is stronger and more diffuse. (Original magnification × 63; scale bar 10μm). Journal of Investigative Dermatology 2010 130, 2440-2447DOI: (10.1038/jid.2010.141) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Amplification plots of real-time RT-PCR assay for tyrosinase mRNA expression of SK-MEL-28 melanoma cell lines and melanoma patients’ CTC isolated on ISET filters. Evaluation of tyrosinase mRNA on selected ISET-positive filters was obtained after direct lysis of cellular material on the spot filter surface and subsequent RNA extraction. Journal of Investigative Dermatology 2010 130, 2440-2447DOI: (10.1038/jid.2010.141) Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions