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[Cancer Research 64, 4099-4104, June 15, 2004]
© 2004 American Association for Cancer Research

Regular Articles

A Technical Triade for Proteomic Identification and Characterization of Cancer Biomarkers

Christian Melle1, Günther Ernst1, Bettina Schimmel1, Annett Bleul1, Sven Koscielny2, Andreas Wiesner4, Ralf Bogumil4, Ursula Möller3, Dirk Osterloh5, Karl-Jürgen Halbhuber3 and Ferdinand von Eggeling1

1 Core Unit Chip Application, Institute of Human Genetics and Anthropology, 2 Clinic of ENT Diseases, and 3 Institute of Anatomy II, Friedrich-Schiller-University, Jena; 4 Ciphergen Biosystems GmbH, Göttingen; and 5 SIRS-Lab GmbH, Jena, Germany

Biomarkers are needed to elucidate the biological background and to improve the detection of cancer. Therefore, we have analyzed laser-microdissected cryostat sections from head and neck tumors and adjacent mucosa on ProteinChip arrays. Two differentially expressed proteins (P = 3.34 x 10–5 and 4.6 x 10–5) were isolated by two-dimensional gel electrophoresis and identified as S100A8 (calgranulin A) and S100A9 (calgranulin B) by in-gel proteolytic digestion, peptide mapping, tandem mass spectrometry analysis, and immunodepletion assay. The relevance of these single marker proteins was evaluated by immunohistochemistry. Positive tissue areas were reanalyzed on ProteinChip arrays to confirm the identity of these proteins. As a control, a peak with low P was identified as calgizzarin (S100A11) and characterized in the same way. This technical triade of tissue microdissection, ProteinChip technology, and immunohistochemistry opens up the possibility to find, identify, and characterize tumor relevant biomarkers, which will allow the movement toward the clonal heterogeneity of malignant tumors. Taking this approach, proteins were identified that might be responsible for invasion and metastasis.




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