P147-T Differential Proteome Analysis of Primary and Metastatic Melanoma Using 2DE and LC-MS/MS.

Melanoma causes the highest rate of skin cancer related fatalities. If detected and treated early, melanoma is mostly curable. Unfortunately, once metastasis has occurred melanoma is often fatal. The biochemical pathways involved in the development from primary to metastatic melanoma is an area under intense investigation. A high-throughput proteomics approach has been applied to better understand the processes that underlie tumor formation and progression. Analyses of global melanoma proteome expression patterns are compared between the primary melanoma cell line WM-115 and the metastatic melanoma cell line WM-266-4, where both cell lines derived from the same patient. Total cell lysates were separated by two dimensional-gel electrophoresis (2DE). After protein differential expression analysis, protein spots of interest were excised, digested with trypsin and analyzed by LC-MS/MS. The mass data were searched against an in-house NCBInr database using the Mascot search engine for protein identification. Comparing the spots between gels, 470 spots were matched for both gels, while 109 spots were up-regulated and 15 spots down-regulated in metastatic melanoma. We identified proteins involved in tumor progression, metabolism, signal transduction, DNA binding, as well as structural and heat shock proteins. This study establishes a step forward in the development of the metstatic melanoma protein database, the understanding of the chemical pathways that lead from primary to metastatic melanoma, and identifying new targets for inhibitor development.