CHANGES IN ENERGY METABOLISM OF BREAST CANCER CELLS INDUCED TO EMT BY EGF

Lanfredi, Guilherme P. (Faculdade de Medicina de Ribeirão Preto – USP), Palma, Camila de S. (Faculdade de Medicina de Ribeirão Preto – USP), Dechandt, Carlos R. P. (Faculdade de Ciências Farmacêuticas de RP – USP), Alberici, Luciane C. (Faculdade de Ciênc

Faculdade de Medicina de Ribeirão Preto da Universidade de São PauloTo investigate the deregulation of cellular energetics, a known hallmark of cancer, analyzing glycolysis and oxygen consuption of MCF7 breast cancer cell line induced to EMT (Epithelial-Mesenchymal Transition) by EGF (Epidermal Growth Factor). MCF7 cells cultured in standard MEM condition and treated with EGF (10ng/ml) were evaluated in terms of O2 consumption by a high-resolution respirometry assay. In parallel, mass spectrometric analysis using MRM (Multiple Reaction Monitoring) method was used to quantitate a panel of glycolytic and other proteins of interest through representative tryptic peptides. The comparison of MRM analysis display increased levels of Hexokinase, Phosphofructokinase and Piruvate kinase (irreversible glycolysis steps) and the respirometry assay exhibited raise of O2 consumption indicating greater mitochondrial capability (ETS state) corroborating with citrate synthase activity superior mitochondria quantity in cells induced to EMT by EGF. This study showed changes related to glucose metabolism and mitochondrial respiration revealing new disposition of EMT induced MCF7 cells compared to normally cultured cells and shows the modulation of breast cancer metabolism in our model of cancer progression. In addition, our results indicate evidences of changes in functional level and characterize metastasis in terms of protein level beyond cytoskeleton, junctions, membrane and transcription factors, commonly used to characterize EMT.Breast Cancer, MCF7, EMT, EGF, Mass Spectrometry, MRM, Respirometry Assay.

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