Iterative sorting reveals CD133+and CD133-melanoma cells as phenotypically distinct populations
AuthorGrasso, C; Anaka, M; Hofmann, O; Sompallae, R; Broadley, K; Hide, W; Berridge, MV; Cebon, J; Behren, A; McConnell, MJ
Source TitleBMC Cancer
PublisherBIOMED CENTRAL LTD
Medicine (Austin & Northern Health)
Document TypeJournal Article
CitationsGrasso, C., Anaka, M., Hofmann, O., Sompallae, R., Broadley, K., Hide, W., Berridge, M. V., Cebon, J., Behren, A. & McConnell, M. J. (2016). Iterative sorting reveals CD133+and CD133-melanoma cells as phenotypically distinct populations. BMC CANCER, 16 (1), https://doi.org/10.1186/s12885-016-2759-2.
Access StatusOpen Access
BACKGROUND: The heterogeneity and tumourigenicity of metastatic melanoma is attributed to a cancer stem cell model, with CD133 considered to be a cancer stem cell marker in melanoma as well as other tumours, but its role has remained controversial. METHODS: We iteratively sorted CD133+ and CD133- cells from 3 metastatic melanoma cell lines, and observed tumourigenicity and phenotypic characteristics over 7 generations of serial xeno-transplantation in NOD/SCID mice. RESULTS: We demonstrate that iterative sorting is required to make highly pure populations of CD133+ and CD133- cells from metastatic melanoma, and that these two populations have distinct characteristics not related to the cancer stem cell phenotype. In vitro, gene set enrichment analysis indicated CD133+ cells were related to a proliferative phenotype, whereas CD133- cells were of an invasive phenotype. However, in vivo, serial transplantation of CD133+ and CD133- tumours over 7 generations showed that both populations were equally able to initiate and propagate tumours. Despite this, both populations remained phenotypically distinct, with CD133- cells only able to express CD133 in vivo and not in vitro. Loss of CD133 from the surface of a CD133+ cell was observed in vitro and in vivo, however CD133- cells derived from CD133+ retained the CD133+ phenotype, even in the presence of signals from the tumour microenvironment. CONCLUSION: We show for the first time the necessity of iterative sorting to isolate pure marker-positive and marker-negative populations for comparative studies, and present evidence that despite CD133+ and CD133- cells being equally tumourigenic, they display distinct phenotypic differences, suggesting CD133 may define a distinct lineage in melanoma.
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