Validation of a phosphoprotein array assay for characterization of human tyrosine kinase receptor downstream signaling in breast cancer.
Fiche publication
Date publication
juillet 2009
Auteurs
Membres identifiés du Cancéropôle Est :
Dr LEROUX Agnès, Pr MERLIN Jean-Louis
Tous les auteurs :
Chergui F, Chretien AS, Bouali S, Ramacci C, Rouyer M, Bastogne T, Genin P, Leroux A, Merlin JL
Lien Pubmed
Résumé
BACKGROUND: Human epidermal growth factor receptor (HER) downstream signaling kinases have important effects on tumor response to anti-HER monoclonal antibodies and tyrosine kinase inhibitors. We validated an assay that uses phosphoprotein arrays for measurement of HER downstream signaling functionality in breast carcinomas. METHODS: Using the Bio-Plex(R) phosphoprotein array (BPA), we performed multiplex immunoanalysis to investigate the expression of phosphorylated epidermal growth factor receptor and phosphorylated HER downstream signaling proteins (phosphorylated protein kinase B, phosphorylated glycogen synthase kinase -3beta, phosphorylated P70 ribosomal protein S6 kinase, and phosphorylated extracellular signal regulated kinase 42/44) in 49 frozen specimens of ductal infiltrating breast carcinoma taken at diagnosis. BPA was cross-validated with Western blot analysis. Sample size, homogenicity, tumor content, protein extraction, and monoclonal antibody detection were in accordance with optimized standard operating procedures. RESULTS: Linear regression showed significant quantitative correlations between BPA and Western blot, with regression coefficient values of 0.71-0.87 (P < 0.001). BPA intra- and interassay CVs were
Référence
Clin Chem. 2009 Jul;55(7):1327-36