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Journal for ImmunoTherapy of Cancer

Open Access

New strategy to reveal the black seeds of melanomas (cancer stem cells) with their vulnerable characteristics at cellular and molecular levels

  • Beatrix Kotlan1, 6,
  • Gabriella Liszkay2, 7, 8,
  • Gyorgy Naszados3,
  • Zoltan Dolescall4,
  • Laszlo Toth5,
  • Laszlo Gobor5,
  • Istvan N Vamosy5,
  • Andras Szollar5,
  • Szabolcs Horvath6,
  • Klara Eles6,
  • Orsolya Csuka4, 7,
  • Miklos Kasler7, 8,
  • Maria Godeny3, 7, 8 and
  • Francesco M Marincola9
Journal for ImmunoTherapy of Cancer20131(Suppl 1):P160

Published: 7 November 2013


MelanomaCancer Stem CellMetastatic MelanomaSelection MarkerGene Expression Microarray


A suitable approach to select cancer stem cells (CSC), the black seed of melanomas would enable their characterization and elimination.


Cancerous tissues from primary and metastatic lesions of patients with malignant melanomas (n=150) were investigated by cell cultures and molecular genetics. Double labelled cells were sorted by BD FACSAvia Sorter. Gene expression analysis by Real Time PCR (MYiQTM, BIO-RAD) and RNA microarray (Agilent) has been performed (n=48).


90% of the cell cultures grew and cancer initiating cells could be IF FACS sorted (0,1% - 1 %). Colocalisation of unique GD3 sialilated glycosphingolipids and antiCD20 binding was proved. Characteristic growth pattern, spheroid forming, CSC markers (e.g. CD133, Nestin, ABCB5, CD20 and unique GD3) was observed (Figure 1.a,b,c,d). We found enhanced gene expression of CXCR4 and other markers correlated to metastatic potential, clinical outcome and CSC presence. High throughput gene expression microarray analysis of RNA preparations of punch biopsies and CSC outgrowth are compared.

Figure 1


Unique GD3 sialilated glycosphingolipids with colocalised CD20 proved to be selection markers for CSC in metastatic melanomas. Our strategy paves the way for detection and characterization of cancer stem cells and provides material for therapeutic developments to eliminate the black seeds of melanomas.



Harry J Lloyd Charitable Trust Melanoma Research Award.

Authors’ Affiliations

Molecular Immunology and Toxicology, National Institute of Oncology, Budapest, Hungary
Oncodermatology, National Institute of Oncology, Budapest, Hungary
Radiological Diagnostics, National Institute of Oncology, Budapest, Hungary
Pathogenetics, National Institute of Oncology, Budapest, Hungary
Oncosurgery, National Institute of Oncology, Budapest, Hungary
Surgical and Molecular Tumorpathology, National Institute of Oncology, Budapest, Hungary
Board of Directors, National Institute of Oncology, Budapest, Hungary
Univ Med Pharm, Tirgu Mures, Romania
SIDRA Medical and Research Center, Doha, Qatar


© Kotlan et al; licensee BioMed Central Ltd. 2013

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.