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Biomarker Discovery and Identification in Laser Microdissected Head and Neck Squamous Cell Carcinoma

Escrito por Carla Brito Lopes

Abstract

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 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 and identified as annexin V by in-gel proteolytic digestion, peptide mapping, tandem mass spectrometry 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, two-dimensional gel electrophoresis, and immunohistochemistry. In our experience, however, such studies only make sense if a relatively pure microdissected tumor tissue is used. Only then minute changes in protein expression between normal pharyngeal epithelium and tumor tissue can be detected, and it will become possible to educe a tumor-associated protein pattern that might be used as a marker for tumorigenesis and progression.

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Article first published: June 23, 2003

doi:10.1074/mcp.M300033-MCP200

URL: here

Citation: Melle C, Ernst G, Schimmel B, Bleul A, Koscielny S, Wiesner A, et al. Biomarker Discovery and Identification in Laser Microdissected Head and Neck Squamous Cell Carcinoma with ProteinChip® Technology, Two-dimensional Gel Electrophoresis, Tandem Mass Spectrometry, and Immunohistochemistry. Mol Cell Proteomics. 2003 Jul 1;2 (7 ):443–52.

Copyright: © 2003 by The American Society for Biochemistry and Molecular Biology, Inc. [Reprinted with permission]

 

Sobre o autor

Carla Brito Lopes

Carla nasceu a 14 de Fevereiro de 1977 em Viana do Castelo. Completou o bacharelato em Anatomia Patológica, Citológica e Tanatológica na ESTES-Porto em 1998 e licenciou-se em 2001 pela Escola Superior de Tecnologia da Saúde em Lisboa. Concluiu a certificação em Laboratory Management pela ASCP (American Association of Clinical Pathology) em Setembro de 2016.
Actualmente encontra-se a frequentar mestrado em Genética Molecular e Biomedicina na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa.