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Molecular & Cellular Proteomics 6:2150-2164, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

A Proteomics Analysis of Cell Signaling Alterations in Colorectal Cancer^*,S

Juan Madoz-Gúrpide^,, Marta Cañamero^¶, Lydia Sanchez^||, José Solano^**, Patricia Alfonso and J. Ignacio Casal^,

From the Protein Technology Unit, ^¶ Comparative Pathology Unit, and ^|| Histology and Immunohistochemistry Unit, Biotechnology Programme, Centro Nacional de Investigaciones Oncológicas (CNIO), c/Melchor Fernández Almagro 3, 28029 Madrid, Spain and ^** Servicio de Anatomia Patológica, Hospital Sta. María del Rosell, Cartagena, 30203 Murcia, Spain

To gain further insight into alterations in cellular pathways, tumor profiling, and marker discovery in colorectal cancer (CRC) we used a new antibody microarray specific for cell signaling. Soluble protein extracts were prepared from paired tumor/normal biopsies of 11 patients diagnosed with colorectal carcinoma at different stages; four liver carcinomas were used as a reference. Antibody microarray analysis identified 46 proteins that were differentially expressed between normal colorectal epithelium and adenocarcinoma. These proteins gave a specific signature for CRC, different from other tumors, as well as a panel of novel markers and potential targets for CRC. Twenty-four proteins were validated by using a specific colorectal cancer tissue microarray and immunoblotting analysis. Together with some previously well known deregulated proteins in CRC (β-catenin, c-MYC, or p63), we found new potential markers preferentially expressed in CRC tumors: cytokeratin 13, calcineurin, CHK1, clathrin light chain, MAPK3, phospho-PTK2/focal adhesion kinase (Ser-910), and MDM2. CHK1 antibodies were particularly effective in discriminating between tumoral and normal mucosa in CRC. Moreover a global picture of alterations in signaling pathways in CRC was observed, including a significant up-regulation of different components of the epidermal growth factor receptor and Wnt/β-catenin pathways and the down-regulation of p14^ARF. The experimental approach described here should be applicable to other pathologies and neoplastic processes.

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