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Unformatted text preview: ts suggest that even in normal adult skin melanocytes PAX3 regulates differentiation and proliferation, as it does during development. It seems likely then, that PAX3 would continue to control maintenance of the undifferentiated state, proliferation and migration, throughout melanocyte development and maturation, and melanoma‐genesis, possibly playing a significant role in driving melanoma development and progression. 79 Chapter IV – Medic, Rizos and Ziman, 2011 To clarify its role and identify any potential differences in the functional repertoire of PAX3 between melanocytes and melanoma, we analysed its direct target genes, and assessed their differential activation in each cell type. We show here that PAX3 is similarly involved in promoting a “stem” cell phenotype in both melanoma cells and melanocytes (via Nestin (NES) and SRY (sex determining region Y)‐box 9 (SOX9)). There is however, a hitherto unreported difference in PAX3 regulation of melanoma cell proliferation relative to melanocytes (through Tumour protein D52 (TPD52)). We also confirm that its role in cell survival (via BCL2L1 and Phosphatase and tensin homologue deleted from chromosome 10 (PTEN)) is distinct for melanoma cells. Moreover, while PAX3 commonly regulates cell migration through Melanoma cell adhesion molecule (MCAM) and Chondroitin sulphate proteoglycan 4 (CSPG4) in both melanocytes and melanoma cells, binding and expression of these genes are significantly enhanced in melanoma cells, presumably affecting the functional outcome. Therefore, PAX3 may indeed ‘tick all the boxes’ as an intrinsic factor in melanoma development and progression. Materials and Methods Cell culture Human primary neonatal foreskin melanocytes HEM1455 (Cell Applications) were cultured as previously described . A2058 metastatic melanoma cells (ATCC) were cultured in DMEM medium (Invitrogen) supplemented with 10% FBS (Interpath). Cell cultures were incubated at 37°C in a humidified incubator supplemented with 5% CO2. Immunocytochemistry Immunofluorescent analysis was done as previously...
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This document was uploaded on 03/06/2014 for the course CHEMISTRY 12 at National University of Singapore.
- Spring '14