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Molecular & Cellular Proteomics 6:1170-1182, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Endometrial Carcinoma Biomarker Discovery and Verification Using Differentially Tagged Clinical Samples with Multidimensional Liquid Chromatography and Tandem Mass Spectrometry^*,S

Leroi V. DeSouza^,,¶, Jörg Grigull^||, Shaun Ghanny^,¶,||, Valérie Dubé^**,, Alexander D. Romaschin^,¶¶, Terence J. Colgan^**,¶¶ and K. W. Michael Siu^,,¶,||||

From the Departments of Chemistry, Biology, and ^|| Mathematics and Statistics and ^¶ Centre for Research in Mass Spectrometry, York University, 4700 Keele Street, Toronto, Ontario M2J 1P3, Canada, ^** Pathology and Laboratory Medicine, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada, Division of Clinical Biochemistry, St. Michael's Hospital, 30 Bond Street, Toronto, Ontario M5B 1W8, Canada, and ^¶¶ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5G 1L5, Canada

The utility of differentially expressed proteins discovered and identified in an earlier study (DeSouza, L., Diehl, G., Rodrigues, M. J., Guo, J., Romaschin, A. D., Colgan, T. J., and Siu, K. W. M. (2005) Search for cancer markers from endometrial tissues using differentially labeled tags iTRAQ and cleavable ICAT with multidimensional liquid chromatography and tandem mass spectrometry. J. Proteome Res. 4, 377–386) to discriminate malignant and benign endometrial tissue samples was verified in a 40-sample iTRAQ (isobaric tags for relative and absolute quantitation) labeling study involving normal proliferative and secretory samples and Types I and II endometrial cancer samples. None of these proteins had the sensitivity and specificity to be used individually to discriminate between normal and cancer samples. However, a panel of pyruvate kinase, chaperonin 10, and ₁-antitrypsin achieved the best results with a sensitivity, specificity, predictive value, and positive predictive value of 0.95 each in a logistic regression analysis. In addition, three new potential markers were discovered, whereas two other proteins showed promising trends but were not detected in sufficient numbers of samples to permit statistical validation. Differential expressions of some of these candidate biomarkers were independently verified using immunohistochemistry.

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