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Functional features of cancer stem cells in melanoma cell lines
BACKGROUND: Recent evidence suggests a subset of cells within a tumor with "stem-like" characteristics. These cells are able to transplant tumors in immunodeficient hosts. Distinct from non-malignant stem cells, cancer stem cells (CSC) show low proliferative rates, high self-renewing capac...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3765139/ https://www.ncbi.nlm.nih.gov/pubmed/23915418 http://dx.doi.org/10.1186/1475-2867-13-78 |
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author | Zimmerer, Rüdiger M Korn, Philippe Demougin, Philippe Kampmann, Andreas Kokemüller, Horst Eckardt, André M Gellrich, Nils-Claudius Tavassol, Frank |
author_facet | Zimmerer, Rüdiger M Korn, Philippe Demougin, Philippe Kampmann, Andreas Kokemüller, Horst Eckardt, André M Gellrich, Nils-Claudius Tavassol, Frank |
author_sort | Zimmerer, Rüdiger M |
collection | PubMed |
description | BACKGROUND: Recent evidence suggests a subset of cells within a tumor with "stem-like" characteristics. These cells are able to transplant tumors in immunodeficient hosts. Distinct from non-malignant stem cells, cancer stem cells (CSC) show low proliferative rates, high self-renewing capacity, propensity to differentiate into actively proliferating tumor cells, and resistance to chemotherapy or radiation. They are often characterized by elevated expression of stem cell surface markers, in particular CD133, and sets of differentially expressed stem cell-associated genes. CSC are usually rare in clinical specimens and hardly amenable to functional studies and gene expression profiling. In this study, a panel of heterogenous melanoma cell lines was screened for typical CSC features. METHODS: Nine heterogeneous metastatic melanoma cell lines including D10 and WM115 were studied. Cell lines were phenotyped using flow cytometry and clonogenic assays were performed by limiting dilution analysis on magnetically sorted cells. Spheroidal growth was investigated in pretreated flasks. Gene expression profiles were assessed by using real-time rt-PCR and DNA microarrays. Magnetically sorted tumor cells were subcutaneously injected into the flanks of immunodeficient mice. Comparative immunohistochemistry was performed on xenografts and primary human melanoma sections. RESULTS: D10 cells expressed CD133 with a significantly higher clonogenic capacity as compared to CD133- cells. Na8, D10, and HBL cells formed spheroids on poly-HEMA-coated flasks. D10, Me39, RE, and WM115 cells expressed at least 2 of the 3 regulatory core transcription factors SOX2, NANOG, and OCT4 involved in the maintenance of stemness in mesenchymal stem cells. Gene expression profiling on CD133+ and CD133- D10 cells revealed 68 up- and 47 downregulated genes (+/-1.3 fold). Two genes, MGP and PROM1 (CD133), were outstandingly upregulated. CD133+ D10 cells formed tumors in NSG mice contrary to CD133- cells and CD133 expression was detected in xenografts and primary human melanoma sections using immunohistochemistry. CONCLUSIONS: Established melanoma cell lines exhibit, to variable extents, the typical features of CSCs. The tumorigenic cell line D10, expressing CD133 and growing in spheroids and might qualify as a potential model of melanoma CSCs. |
format | Online Article Text |
id | pubmed-3765139 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-37651392013-09-07 Functional features of cancer stem cells in melanoma cell lines Zimmerer, Rüdiger M Korn, Philippe Demougin, Philippe Kampmann, Andreas Kokemüller, Horst Eckardt, André M Gellrich, Nils-Claudius Tavassol, Frank Cancer Cell Int Primary Research BACKGROUND: Recent evidence suggests a subset of cells within a tumor with "stem-like" characteristics. These cells are able to transplant tumors in immunodeficient hosts. Distinct from non-malignant stem cells, cancer stem cells (CSC) show low proliferative rates, high self-renewing capacity, propensity to differentiate into actively proliferating tumor cells, and resistance to chemotherapy or radiation. They are often characterized by elevated expression of stem cell surface markers, in particular CD133, and sets of differentially expressed stem cell-associated genes. CSC are usually rare in clinical specimens and hardly amenable to functional studies and gene expression profiling. In this study, a panel of heterogenous melanoma cell lines was screened for typical CSC features. METHODS: Nine heterogeneous metastatic melanoma cell lines including D10 and WM115 were studied. Cell lines were phenotyped using flow cytometry and clonogenic assays were performed by limiting dilution analysis on magnetically sorted cells. Spheroidal growth was investigated in pretreated flasks. Gene expression profiles were assessed by using real-time rt-PCR and DNA microarrays. Magnetically sorted tumor cells were subcutaneously injected into the flanks of immunodeficient mice. Comparative immunohistochemistry was performed on xenografts and primary human melanoma sections. RESULTS: D10 cells expressed CD133 with a significantly higher clonogenic capacity as compared to CD133- cells. Na8, D10, and HBL cells formed spheroids on poly-HEMA-coated flasks. D10, Me39, RE, and WM115 cells expressed at least 2 of the 3 regulatory core transcription factors SOX2, NANOG, and OCT4 involved in the maintenance of stemness in mesenchymal stem cells. Gene expression profiling on CD133+ and CD133- D10 cells revealed 68 up- and 47 downregulated genes (+/-1.3 fold). Two genes, MGP and PROM1 (CD133), were outstandingly upregulated. CD133+ D10 cells formed tumors in NSG mice contrary to CD133- cells and CD133 expression was detected in xenografts and primary human melanoma sections using immunohistochemistry. CONCLUSIONS: Established melanoma cell lines exhibit, to variable extents, the typical features of CSCs. The tumorigenic cell line D10, expressing CD133 and growing in spheroids and might qualify as a potential model of melanoma CSCs. BioMed Central 2013-08-06 /pmc/articles/PMC3765139/ /pubmed/23915418 http://dx.doi.org/10.1186/1475-2867-13-78 Text en Copyright © 2013 Zimmerer et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Primary Research Zimmerer, Rüdiger M Korn, Philippe Demougin, Philippe Kampmann, Andreas Kokemüller, Horst Eckardt, André M Gellrich, Nils-Claudius Tavassol, Frank Functional features of cancer stem cells in melanoma cell lines |
title | Functional features of cancer stem cells in melanoma cell lines |
title_full | Functional features of cancer stem cells in melanoma cell lines |
title_fullStr | Functional features of cancer stem cells in melanoma cell lines |
title_full_unstemmed | Functional features of cancer stem cells in melanoma cell lines |
title_short | Functional features of cancer stem cells in melanoma cell lines |
title_sort | functional features of cancer stem cells in melanoma cell lines |
topic | Primary Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3765139/ https://www.ncbi.nlm.nih.gov/pubmed/23915418 http://dx.doi.org/10.1186/1475-2867-13-78 |
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