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E-GEOD-48838 - Side population cells in human primary melanoma

Released on 27 May 2014, last updated on 3 June 2014
Homo sapiens
Samples (8)
Array (1)
Protocols (6)
Melanoma remains the most lethal skin cancer, mainly because of high resistance to therapy. Side population (SP) cells are found in many types of cancer and are usually enriched in therapy-resistant as well as tumorigenic cells. Here, we identified a Hoechst dye-effluxing SP in a large series of human melanoma samples representing different progression phases. The SP size did not change with disease stage but was correlated with the prognostic “Breslow’s depth” in the primary (cutaneous) tumors. When injected into immunodeficient mice, the SP generated larger tumors than the bulk “main population” (MP) melanoma cells in two consecutive generations, and showed tumorigenic capacity at lower cell numbers than the MP. In addition, the SP reconstituted the heterogeneous composition of the human A375 melanoma cell line, and its clonogenic activity was 2.5-fold higher than that of the MP. Gene-expression analysis revealed upregulated expression in the melanoma SP (versus the MP) of genes associated with chemoresistance and anti-apoptosis. Consistent with these molecular characteristics, the SP increased in proportion when A375 cells were exposed to the melanoma standard chemotherapeutic agent dacarbazine, and to the aggravating condition of hypoxia. In addition, the SP showed enhanced expression of genes related to cell invasion and migration, as well as to putative (melanoma) cancer stem cells (CSC) including ABCB1 and JARID1B. ABCB1 immunoreactivity was detected in a number of tumor cells in human melanomas, and in particular in clusters at the invasive front of the primary tumors. Together, our findings support that the human melanoma SP is enriched in tumorigenic and chemoresistant capacity, considered key characteristics of CSC. The melanoma SP may therefore represent an interesting therapeutic target. Four primary melanomas (subtype SSMM) were dissociated into single cells. From each of these single cell suspensions, side population (SP) and bulk \main population\ (MP) cells were isolated based on their Hoechst-effluxing capability using fluorescence-activated cell sorting. Gene expression of SP cells is analyzed relative to the gene expression of the corresponding MP cells.
Experiment type
transcription profiling by array 
Wouter Van Delm <>, Anke Vandenbroeck, Hannelore Maes, Hugo Vankelecom, Jasper Wouters, Joost vandenOord, Lies Gremeaux, Marguerite Stas, Olivier Govaere, Patrizia Agostinis, Tania Roskams
The human melanoma side population displays molecular and functional characteristics of enriched chemoresistance and tumorigenesis. Wouters J, Stas M, Gremeaux L, Govaere O, Van den Broeck A, Maes H, Agostinis P, Roskams T, van den Oord JJ, Vankelecom H. , PMID:24098529
Investigation descriptionE-GEOD-48838.idf.txt
Sample and data relationshipE-GEOD-48838.sdrf.txt
Raw data (1)
Processed data (1)
Array designA-AGIL-28.adf.txt