Protein microarrays for the detection of biomarkers in head and neck squamous cell carcinomas.
Protein microarrays are of increasing importance for high-throughput screening of fresh tissues. In our study, protein microarrays were generated by printing antibodies onto membranes to characterize protein profiles expressed by head and neck squamous cell carcinomas (HNSCCs). Cellular proteomes of 30 matched normal squamous epithelial cells and carcinoma specimens were analyzed after tissue microdissection using microarrays composed of 83 different antibodies. As controls, Western blot analysis and tissue microarrays (TMAs) containing 98 HNSCC specimens were used. Of the 83 proteins examined, 14 showed differential expression between HNSCCs and normal epithelium. The protein microarray approach revealed an upregulation of 8 proteins and a downregulation of 6 proteins. Bag-1, Cox-2, Hsp-70, Stat3, pescadillo, MMP-7 (matrilysin), IGF-2, and cyclin D1 were identified to be significantly upregulated, whereas suppressor of cytokine signaling 1, thrombospondin, TGF-beta1, Jun, Fos, and Fra-2 were downregulated. The differential expression of these proteins was confirmed using Western blot and TMA. Upon correlation of differentially regulated proteins with the clinicopathologic data of our patients, MMP-7 (matrilysin) was found to be associated with survival in univariate, but not multivariate, analysis. These data indicate that our protein arrays provide protein information in a systematic, reproducible, and also high-throughput fashion.
Weber
Anette
A
Department of ENT, University of Leipzig, 04103 Leipzig, Germany.
Hengge
Ulrich R
UR
Stricker
Ingo
I
Tischoff
Iris
I
Markwart
Annett
A
Anhalt
Kathrin
K
Dietz
Andreas
A
Wittekind
Christian
C
Tannapfel
Andrea
A
D014408:Biomarkers, Tumor / Q000032:analysis*; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology*; D019938:Cyclin D1 / Q000032:analysis; D006258:Head and Neck Neoplasms / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D020783:Matrix Metalloproteinase 7 / Q000032:analysis; D015999:Multivariate Analysis; D040081:Protein Array Analysis / Q000379:methods*; D020543:Proteome / Q000032:analysis; D015203:Reproducibility of Results; D050796:STAT3 Transcription Factor / Q000032:analysis; D046888:Tissue Array Analysis
D014408:Biomarkers, Tumor; D020543:Proteome; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D019938:Cyclin D1; C060932:MMP7 protein, human; D020783:Matrix Metalloproteinase 7