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

BMC Cancer

Research Article

Ramakrishna Sompallae¹  Oliver Hofmann²  Winston Hide³  Jonathan Cebon⁴  Matthew Anaka⁴  Andreas Behren⁴  Kate Broadley⁵  Michael V. Berridge⁵  Carole Grasso⁵  Melanie J. McConnell⁵ 

[1] Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, 02115, Boston, MA, USA;Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, 02115, Boston, MA, USA;Harvard Stem Cell Institute, Holyoke Center, Suite 727W, 1350 Massachusetts Avenue, 02138, Cambridge, MA, USA;Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, 02115, Boston, MA, USA;Sheffield Institute for Translational Neuroscience, The University of Sheffield, 385a Glossop Road, S10 2HQ, Sheffield, UK;Ludwig Institute for Cancer Research, Olivia Newton-John Cancer & Wellness Centre, Austin Hospital, 3084, Heidelberg, VIC, Australia;Malaghan Institute of Medical Research, P.O. Box 7060, 6242, Wellington, New Zealand;

关键词: Melanoma;    Melanoma Cell;    Cancer Stem Cell;    Cancer Stem Cell Marker;    Cell Surface CD133;   

DOI  :  10.1186/s12885-016-2759-2

 received in 2015-06-24, accepted in 2016-09-02,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundThe 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.MethodsWe 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.ResultsWe 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.ConclusionWe 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.

【 授权许可】

CC BY   
© The Author(s). 2016