Purpose To further the molecular biological understanding of the events governing the development of metastatic disease and to identify potential targets for therapy.
Methods 8 metastatic vs 8 non‐metastatic fresh uveal melanoma tissue with a minimum follow‐up of 10 years were subjected to quantitative, label‐free proteomic analysis. Strict criteria was applied in the Progenesis LC‐MS software to filter out statistically non‐significant peptides and proteins. These were anova p < 0.05, power > 0.8 and protein confidence score > 100. Differential expression of proteins and their relative abundance in the 2 disease groups were determined using the same software.
Results 653 proteins were identified (global proteome of all 16 samples). Of these, 61 proteins met the statistics criteria applied. 9 of those had good separation between metastatic and non‐metastatic disease groups. 4 proteins were selected for further follow‐up and validation. Currently, immunohistochemistry studies are being performed to confirm expression levels in paraffin embedded tissue slides. Then, siRNA knockdown and immunoprecipitation experiments will be performed in uveal melanoma cell lines (primary and metastatic) to determine downstream effects and potential targets for therapy.
Conclusion Proteomic analysis of 8 metastatic vs 8 non‐metastatic fresh uveal melanoma tissue has identified 4 proteins which are currently being validated using immunohistochemistry, siRNA knockdown and immunoprecipitation experiments.