>>Mol Cell Proteomics. 2003 Jun 23
Biomarker discovery and identification in laser microdissected head and neck squamous cell carcinoma with ProteinChip(R) technology, two-dimensional gel electrophoresis, tandem mass spectrometry and immunohistochemistry.
Melle C, Ernst G, Schimmel B, Bleul A, Koscielny S, Wiesner A, Bogumil R, Moller U, Osterloh D, Halbhuber KJ, Von Eggeling F.
Klinikum der FSU, Institut fr Humangenetik, Core Unit Chip Application (CUCA), Jena 07740.
Head and neck cancer is a frequent malignancy with a complex, and up to now not clear etiology. Therefore, despite of improvements in diagnosis and therapy, the survival rate with head and neck squamous-cell carcinomas (HNSCC) is poor. For a better understanding of the molecular mechanisms behind the process of tumorigenesis and tumor progression we have analyzed changes of protein expression between microdissected normal pharyngeal epithelium and tumor tissue by ProteinChip technology. For this cryostat sections from head and neck tumors (n = 57) and adjacent mucosa (n = 44) were laser-microdissected and analyzed on ProteinChip Arrays. The derived mass spectrometry profiles exhibited numerous statistical differences. One peak significantly higher expressed in the tumor (p = 0.000029) was isolated by two-dimensional gel electrophoresis (2-DE) and identified as annexin V by in-gel proteolytic digestion, peptide mapping, Tandem MS/MS analysis and immuno-deplete assay. The relevance of this single marker protein was further evaluated by immunohistochemistry. Annexin-positive tissue areas were re-analyzed on ProteinChip Arrays to confirm the identity of this protein. In this study we could show that biomarker in head and neck cancer can be found, identified and assessed by combination of ProteinChip technology, 2-DE, and immunohistochemistry. In our experience, however, such studies only make sense if a relatively pure microdissected tumor tissue is used. Only then can minute changes in protein expression between normal pharyngeal epithelium and tumor tissue be detected and it will become possible to educe tumor associated protein pattern that might be used as a marker for tumorigenesis and progression. Acknowledgement: This work was support by a grant of the German Federal Ministry of Education and Research (BMBF) and the Interdisciplinary Center for Clinical Research (ICCR), Jena.<<
Cheers, Tuck |
