Discovery and validation of colonic tumor-associated proteins via metabolic labeling and stable isotopic dilution
The unique biology of a neoplasm is reflected by its distinct molecular profile compared with normal tissue. To understand tumor development better, we have undertaken a quantitative proteomic search for abnormally expressed proteins in colonic tumors from ApcMin/⁺ (Min) mice. By raising pairs of Mi...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 106; no. 40; pp. 17235 - 17240 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
National Academy of Sciences
06.10.2009
National Acad Sciences |
Subjects | |
Online Access | Get full text |
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Summary: | The unique biology of a neoplasm is reflected by its distinct molecular profile compared with normal tissue. To understand tumor development better, we have undertaken a quantitative proteomic search for abnormally expressed proteins in colonic tumors from ApcMin/⁺ (Min) mice. By raising pairs of Min and wild-type mice on diets derived from natural-abundance or ¹⁵N-labeled algae, we used metabolic labeling to compare protein levels in colonic tumor versus normal tissue. Because metabolic labeling allows internal control throughout sample preparation and analysis, technical error is minimized as compared with in vitro labeling. Several proteins displayed altered expression, and a subset was validated via stable isotopic dilution using synthetic peptide standards. We also compared gene and protein expression among tumor and nontumor tissue, revealing limited correlation. This divergence was especially pronounced for species showing biological change, highlighting the complementary perspectives provided by transcriptomics and proteomics. Our work demonstrates the power of metabolic labeling combined with stable isotopic dilution as an integrated strategy for the identification and validation of differentially expressed proteins using rodent models of human disease. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 ObjectType-Undefined-3 Author contributions: E.L.H., X.C., A.D.H., W.F.D., and M.R.S. designed research; E.L.H. and X.C. performed research; E.L.H., G.A.B.-W., R.B.H., and A.C.H. contributed new reagents/analytic tools; E.L.H. and M.A.N. analyzed data; and E.L.H., W.F.D., and M.R.S. wrote the paper. 2Present address: Department of Medicine, University of Wisconsin, Madison, WI 53792. 1Present address: Department of Cell Biology, Harvard Medical School, Boston, MA 02115. Contributed by William F. Dove, August 14, 2009 |
ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.0909282106 |