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A sensitive enzyme-linked immunosorbent assay used for quantitation of epidermal growth factor receptor protein in
M E Christensen1, F Engbaek, M H Therkildsen
1Department of Oto-Laryngology-Head and Neck Surgery, Rigshospitalet, University of Copenhagen, Denmark.
Abstract:
The EGF receptor is a transmembrane glycoprotein exerting mitogenic effects on epithelial cells. The purpose of the present study was to develop a sensitive enzyme-linked immunosorbent assay (ELISA) for determination of the epidermal growth factor receptor (EGFR) protein to examine whether the receptor was overexpressed in head and neck squamous cell carcinomas compared with the normal counterpart, and to establish whether clinicopathological correlations were present by investigating a broad spectrum of parameters (tumour size, clinical stage, positive lymph nodes, tumour site, histological grade, keratinisation, preoperative irradiation and clinical outcome). The assay employs two commercially available monoclonal antibodies, both detecting protein epitopes. The material comprises 60 head and neck carcinomas, corresponding normal tissue and normal oral mucosa from healthy individuals. The study demonstrates significantly higher receptor levels in tumours compared with normal tissue (P < 0.002) and a range in tumours and normal tissues of 0.4-10.5 and 0.1-4.3 nmol g-1 membrane protein respectively. Quantitation of receptors in normal mucosa emphasises the importance of using the patients' corresponding normal tissue, because using the patients' mucosa resulted in 83% overexpression, while using normal mucosa from healthy individuals only demonstrated overexpression in 50% of cases. No significant clinicopathological correlations could be established, although the mean values for EGFR increased with tumour size and advanced clinical stage. Furthermore, the prognostic value concerning disease-free survival, recurrence and the time interval for recurrence were investigated but no significance could be demonstrated. In conclusion, the investigation supports the theory of overexpression of EGFR protein as a common motif for malignant epithelial tumours, but limitations in interpretations are demonstrated and discussed further.