Functional features of cancer stem cells in melanoma cell lines

Cancer Cell International

Frank Tavassol¹  Nils-Claudius Gellrich¹  André M Eckardt¹  Horst Kokemüller¹  Andreas Kampmann¹  Philippe Demougin²  Philippe Korn¹  Rüdiger M Zimmerer¹ 

[1] Department of Oral and Maxillofacial Surgery, Head & Neck Oncology Laboratory, Hannover Medical School, Carl-Neuberg-Str. 1, Hannover D-30625, Germany;Division of Molecular Psychology, Life Sciences Training Facility (LSTF), Missionsstrasse 60/62, Basel CH-4055, Switzerland

关键词: WM115;    Melanoma;    D10;    Cell lines;    CD271;    CD146;    CD133;    CD117;    Cancer stem cells;   

Others  :  793190
DOI  :  10.1186/1475-2867-13-78

 received in 2013-03-21, accepted in 2013-08-01,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Zimmerer et al.; licensee BioMed Central Ltd.

【 图 表 】

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