A technical triade for proteomic identification and characterization of cancer biomarkers.
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.
Melle
Christian
C
Core Unit Chip Application, Institute of Human Genetics and Anthropology, Friedrich-Schiller-University, Jena, Germany.
Ernst
Günther
G
Schimmel
Bettina
B
Bleul
Annett
A
Koscielny
Sven
S
Wiesner
Andreas
A
Bogumil
Ralf
R
Möller
Ursula
U
Osterloh
Dirk
D
Halbhuber
Karl-Jürgen
KJ
von Eggeling
Ferdinand
F
D014408:Biomarkers, Tumor / Q000378:metabolism*; D040501:Calgranulin A / Q000378:metabolism; D040502:Calgranulin B / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D016616:Cryoultramicrotomy; D004848:Epithelium / Q000378:metabolism; D006258:Head and Neck Neoplasms / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D010614:Pharynx / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods; D040901:Proteomics / Q000379:methods*; D009418:S100 Proteins / Q000378:metabolism
D014408:Biomarkers, Tumor; D040501:Calgranulin A; D040502:Calgranulin B; D009418:S100 Proteins; C072337:S100A11 protein, human