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    Iterative sorting reveals CD133+and CD133-melanoma cells as phenotypically distinct populations

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    7
    Author
    Grasso, C; Anaka, M; Hofmann, O; Sompallae, R; Broadley, K; Hide, W; Berridge, MV; Cebon, J; Behren, A; McConnell, MJ
    Date
    2016-09-09
    Source Title
    BMC Cancer
    Publisher
    BIOMED CENTRAL LTD
    University of Melbourne Author/s
    Behren, Andreas; CEBON, JONATHAN; ANAKA, MATTHEW; Hofmann, Oliver
    Affiliation
    Medicine and Radiology
    Clinical Pathology
    Metadata
    Show full item record
    Document Type
    Journal Article
    Citations
    Grasso, 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 Status
    Open Access
    URI
    http://hdl.handle.net/11343/260207
    DOI
    10.1186/s12885-016-2759-2
    Abstract
    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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