Application of 2D-DIGE to formalin-fixed diseased tissue samples from hospital repositories: Results from four case studies

Alessandro Tanca, Salvatore Pisanu, Grazia Biosa, Daniela Pagnozzi, Elisabetta Antuofermo, Giovanni P. Burrai, Vincenzo Canzonieri, Paolo Cossu-Rocca, Valli De Re, Albino Eccher, Giuseppe Fanciulli, Stefano Rocca, Sergio Uzzau, Maria Filippa Addis

Research output: Contribution to journalArticle

Abstract

Purpose: In the recent past, the potential suitability of fixed samples to 2D-DIGE studies has been demonstrated on model tissues, but not on "real-world" archival tissues. Therefore, this study was aimed to assess the quality of the results delivered by 2D-DIGE on samples retrieved from hospital tissue repositories. Experimental design: Diseased and normal tissue samples (namely, human gastric adenocarcinoma and normal gastric tissue, human lung neuroendocrine tumors, canine mammary tubulo-papillary carcinoma and normal mammary tissue, sheep liver with cloudy swelling degeneration and normal liver tissue) were retrieved from human and veterinary biorepositories and subjected to full-length protein extraction, cyanine labeling, 2D-DIGE separation, image analysis, MS analysis, and protein identification. Results: Archival samples could be successfully subjected to 2D-DIGE, providing maps of satisfactory resolution, although with varying pattern complexity (possibly influenced by preanalytical variables). Moreover, differentially expressed protein identities were consistent with the disease biology. Conclusions and clinical relevance: 2D-DIGE can support biomarker discovery and validation studies on large sample cohorts. In fact, although some information complexity is lost when compared to fresh-frozen tissues, their vast availability and the associated patient information can considerably boost studies suffering limited sample availability or involving long-distance exchange of samples.

Original languageEnglish
Pages (from-to)252-263
Number of pages12
JournalProteomics - Clinical Applications
Volume7
Issue number3-4
DOIs
Publication statusPublished - Apr 2013

Keywords

  • Canine mammary tumor
  • FFPE
  • Gastric adenocarcinoma
  • Gel-based proteomics
  • Lung neuroendocrine tumor

ASJC Scopus subject areas

  • Clinical Biochemistry

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    Tanca, A., Pisanu, S., Biosa, G., Pagnozzi, D., Antuofermo, E., Burrai, G. P., Canzonieri, V., Cossu-Rocca, P., Re, V. D., Eccher, A., Fanciulli, G., Rocca, S., Uzzau, S., & Addis, M. F. (2013). Application of 2D-DIGE to formalin-fixed diseased tissue samples from hospital repositories: Results from four case studies. Proteomics - Clinical Applications, 7(3-4), 252-263. https://doi.org/10.1002/prca.201200054