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Profiling signatures of ovarian cancer tumour suppression using 2D-DIGE and 2D-LC-MS/MS with tandem mass tagging.
Epithelial ovarian cancer (EOC) is the most common form of gynaecological malignancy in the developed world and has a poor prognosis due to its late detection. Identifying molecular markers of the disease may provide novel approaches to screening and could enable targeted treatment and the design of novel therapies. Although blood is recognized as a highly important source of disease-related biomarkers, the complexity and dynamic range of protein abundance in body fluids has hampered proteomic biomarker discovery and alternative approaches using cell models may be more successful. Herein, we have utilized two cellular models of EOC, where transfer of normal chromosome 18 material into the EOC cell lines TOV-112D and TOV-21G induced in vitro and in vivo suppression of their tumourigenic phenotype. A combination of quantitative two-dimensional difference gel electrophoresis (2D-DIGE) and two-dimensional-liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) with tandem mass tagging (TMT) was employed to examine the whole cell, secreted and crude membrane proteomes of the parental and hybrid cell models to identify differentially expressed proteins as potential markers of tumour suppression. Protein changes of interest were confirmed by immunoblotting in additional hybrid and revertant cell lines where incorporated chromosome 18 material was lost. One candidate marker was also tested in sera from a set of ovarian cancer cases and controls. We have identified a list of promising candidate biomarkers for further testing and functional characterization.
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