TY - JOUR
T1 - Application of 2D-DIGE to formalin-fixed diseased tissue samples from hospital repositories
T2 - Results from four case studies
AU - Tanca, Alessandro
AU - Pisanu, Salvatore
AU - Biosa, Grazia
AU - Pagnozzi, Daniela
AU - Antuofermo, Elisabetta
AU - Burrai, Giovanni P.
AU - Canzonieri, Vincenzo
AU - Cossu-Rocca, Paolo
AU - Re, Valli De
AU - Eccher, Albino
AU - Fanciulli, Giuseppe
AU - Rocca, Stefano
AU - Uzzau, Sergio
AU - Addis, Maria Filippa
PY - 2013/4
Y1 - 2013/4
N2 - 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.
AB - 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.
KW - Canine mammary tumor
KW - FFPE
KW - Gastric adenocarcinoma
KW - Gel-based proteomics
KW - Lung neuroendocrine tumor
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U2 - 10.1002/prca.201200054
DO - 10.1002/prca.201200054
M3 - Article
C2 - 23090899
AN - SCOPUS:84876338400
VL - 7
SP - 252
EP - 263
JO - Proteomics - Clinical Applications
JF - Proteomics - Clinical Applications
SN - 1862-8346
IS - 3-4
ER -