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Originally published In Press as doi:10.1074/mcp.M600175-MCP200 on July 8, 2006.
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Molecular & Cellular Proteomics 6:987-999, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Research

Comparative Proteomics Analysis of Barrett Metaplasia and Esophageal Adenocarcinoma Using Two-dimensional Liquid Mass Mapping*,S

Jia Zhao{ddagger}, Andrew C. Chang§,¶, Chen Li{ddagger}, Kerby A. Shedden||, Dafydd G. Thomas**, David E. Misek§, Arun Prasad Manoharan{ddagger}{ddagger}, Thomas J. Giordano,**, David G. Beer§,¶ and David M. Lubman{ddagger},§,¶,§§

From the Departments of {ddagger} Chemistry, || Statistics, and {ddagger}{ddagger} Bioinformatics, University of Michigan and Departments of § Surgery and ** Pathology and Comprehensive Cancer Center, University of Michigan Medical Center, Ann Arbor, Michigan 48109

Esophageal adenocarcinoma, currently the seventh leading cause of cancer-related death, has been associated with the presence of Barrett metaplasia. The malignant potential of Barrett metaplasia is evidenced by ultimate progression of this condition to invasive adenocarcinoma. We utilized liquid phase separation of proteins with chromatofocusing in the first dimension and nonporous reverse phase HPLC in the second dimension followed by ESI-TOF mass spectrometry to identify proteins differentially expressed in six Barrett metaplasia samples as compared with six esophageal adenocarcinoma samples; all six Barrett samples were obtained from the identical six patients from whom we obtained the esophageal adenocarcinoma tissue. Approximately 300 protein bands were detected by mass mappings, and 38 differentially expressed proteins were identified by µLC-MS/MS. The false positive rates of the peptide identifications were evaluated by reversed database searching. Among the proteins that were identified, Rho GDP dissociation inhibitor 2, {alpha}-enolase, Lamin A/C, and nucleoside-diphosphate kinase A were demonstrated to be up-regulated in both mRNA and protein expression in esophageal adenocarcinomas relative to Barrett metaplasia. Candidate proteins were examined at the mRNA level using high density oligonucleotide microarrays. The cellular expression patterns were verified in both esophageal adenocarcinomas and in Barrett metaplasia by immunohistochemistry. These differentially expressed proteins may have utility as useful candidate markers of esophageal adenocarcinoma.

§§ To whom correspondence should be addressed: Dept. of Surgery, University of Michigan, Ann Arbor, MI 48109. Tel.: 734-647-8834; Fax: 734-615-2088; E-mail: dmlubman{at}umich.edu


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