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Wendy w. Gao, John lattier, pingbo liu, hua yang, martina herwig, Hans E. Grossniklaus; Characterizing Cancer Stem Cells In Uveal Melanoma Cell Lines. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3389.
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© ARVO (1962-2015); The Authors (2016-present)
Previous work has shown evidence of ocular melanoma stem-like cells (Kalirai et al IOVS 2011;52:8458). Subgroups of cancer stem cells are capable of self-renewal, multipotency and differentiation plasticity and treatment resistance. This study analyzed phenotype expression of cancer stem cell markers and migration /invasion properties of melanoma spheres from uveal melanoma cell lines.
Uveal melanoma cell lines from the primary tumor (Mel 270 and Mel 290) were used. None adherent melanoma spheres culture and colony forming efficiency assays were performed to assess the self-renewal. Expression of differentiation and putative stem cell markers were analyzed on both MS and adherent monolayer cells by immunofluorescence. Migration/invasion assays were compared between melanoma spheres and adherent monolayer cells. Live cell imaging of melanoma cells from 3-D collagen cell culture and H&E staining of melanoma stem cells (MSCs) were performed to assess the morphology, proliferation and migration.
Live cell imaging of uveal melanoma cells demonstrated morphologic, proliferative and migratory heterogeneity. A population of small cells exhibited high rates of proliferation and migration compared to other cell types. Both cell lines were capable of forming melanoma spheres at clonal density under non-adhesive cell culture conditions, although non-stem cells adhered and formed adherent monolayer under adhesive cell culture conditions. These melanoma spheres could be passaged to generate further spheres, suggesting self-renewal capacity and enrichment of MSCs. The average colony forming efficiency of Mel 270 and Mel 290 were 0.02% and 0.04% respectively. Expression of CD133, CD271, C-kit, and MITF was greater in MSCs compared to adhesive non-stem cells; Nestin and HMB45 were expressed in both groups, but the level is higher in adhesive non-stem cells. The above markers’ expression indicated the neuroectodermal origin and differentiation properties of MSCs. Migration and invasion assays showed that the invasion rate of melanoma spheres was significantly higher than that of adhesive monolayer cells (t=3.3x10-6, p<0.001). H&E staining of melanoma spheres helped to identify the small MSCs.
This study adds evidence to the existence of MSCs in uveal melanoma cell line. This subgroup of cells exhibits a greater migration and invasion attributes than non-stem cells.
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