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Proteomic comparison of colorectal tumours and non-neoplastic mucosa from paired patient samples using iTRAQ mass spectrometry


Quantitative mass spectrometry using iTRAQ was used to identify differentially expressed proteins from 16 colorectal cancer (CRC) tumours compared to patient-paired adjacent normal mucosa. Over 1400 proteins were identified and quantitated, with 118 determined as differentially expressed by >1.3-fold, with false discovery rate < 0.05. Gene Ontology analysis indicated that proteins with increased expression levels in CRC tumours include those associated with glycolysis, calcium binding, and protease inhibition. Proteins with reduced levels in CRC tumours were associated with loss of ATP production through: (i) reduced beta-oxidation of fatty acids, (ii) reduced NADH production by the tricarboxylic acid cycle and (iii) decreased oxidative phosphorylation activity. Additionally, biosyntheses of glycosaminoglycans and proteoglycans were significantly reduced in tumour samples. Validation experiments using immunoblotting and immunohistochemistry (IHC) showed strong concordance with iTRAQ data suggesting that this workflow is suitable for identifying biomarker candidates. We discuss the uses and challenges of this approach to generate biomarker leads for patient prognostication.

Type Journal
ISBN 1742-2051 (Electronic) 1742-2051 (Linking)
Authors Jankova, L.; Chan, C.; Fung, C. L.; Song, X.; Kwun, S. Y.; Cowley, M. J.; Kaplan, W.; Dent, O. F.; Bokey, E. L.; Chapuis, P. H.; Baker, M. S.; Robertson, G. R.; Clarke, S. J.; Molloy, M. P.;
Publisher Name Molecular BioSystems
Published Date 2011-08-03
Published Volume 7
Published Issue 11
Published Pages 2997-3005
Status Published in-print
URL link to publisher's version
OpenAccess link to author's accepted manuscript version