Identification of novel proteins associated with hepatocellular carcinomas using protein microarrays.
Characterization of the protein profiles expressed by hepatocellular carcinomas (HCCs) may identify the genes involved in hepatocellular carcinogenesis and offers the possibility of elucidating clinical biomarkers. In an effort to discover such proteins and pathways that are deregulated in hepatocellular carcinogenesis, cellular proteomes of matched normal liver cells and carcinoma were analysed by tissue microdissection and protein microarrays. Using protein microarrays made up of 83 different antibodies, it was possible to monitor alterations of the protein levels in HCC and non-neoplastic liver tissue. Further analysis of altered proteins was performed using western blot analysis and tissue microarrays (TMAs) containing 210 HCC specimens and corresponding liver tissue. The protein microarray approach revealed differential expression between HCC and normal liver of 32 of the 83 proteins examined: 21 of these were up-regulated and 11 down-regulated. IGF (insulin growth factor) II, ADAM (a disintegrin and metalloproteases) 9, STAT (signal transducers and activators of transcription) 3, SOCS (suppressors of cytokine signalling) 3, and cyclin D1 were significantly up-regulated and collagen I, SMAD 4, FHIT (fragile histidine triad), and SOCS1 were down-regulated. The differential expression of these proteins was confirmed using western blot analysis and TMAs. Correlation of differentially regulated proteins with clinico-pathological data showed that cyclin D1 and SOCS1 were associated with tumour prognosis in univariate analysis, but not multivariate analysis. These data indicate that the development of an array-based approach for the determination of protein profiles in HCC may facilitate the identification of new proteins associated with carcinogenesis or prognosis.
D051722:ADAM Proteins; D017766:Acid Anhydride Hydrolases; D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western / Q000379:methods; D006528:Carcinoma, Hepatocellular / Q000175:diagnosis*; D002352:Carrier Proteins / Q000235:genetics; D024042:Collagen Type I / Q000235:genetics; D019938:Cyclin D1 / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D019483:Disintegrins / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D008099:Liver / Q000378:metabolism; D008113:Liver Neoplasms / Q000175:diagnosis*; D008565:Membrane Proteins; D008666:Metalloendopeptidases / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics*; D009367:Neoplasm Staging; D040081:Protein Array Analysis; D011506:Proteins / Q000235:genetics; D012097:Repressor Proteins; D015203:Reproducibility of Results; D050796:STAT3 Transcription Factor; D051901:Smad4 Protein; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors; D000099170:Bis(5'-adenosyl)triphosphatase
D051722:ADAM Proteins; D017766:Acid Anhydride Hydrolases; D014408:Biomarkers, Tumor; D002352:Carrier Proteins; D024042:Collagen Type I; D019938:Cyclin D1; D004268:DNA-Binding Proteins; D019483:Disintegrins; D007335:Insulin-Like Growth Factor II; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008666:Metalloendopeptidases; D009363:Neoplasm Proteins; D011506:Proteins; D012097:Repressor Proteins; D050796:STAT3 Transcription Factor; D051901:Smad4 Protein; D000071222:Suppressor of Cytokine Signaling 1 Protein; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D000099170:Bis(5'-adenosyl)triphosphatase; C497796:SMAD4 protein, human; C108944:SOCS1 protein, human; C106644:SOCS3 protein, human; C494086:STAT3 protein, human; C098339:ADAM9 protein, human