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Biotechnology and Applied Biochemistry (2007) 48, (21–27) (Printed in Great Britain)

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Functional expression of human-epidermal-growth-factor receptor in a melanoma cell line

Arlhee Diaz¹, Eduardo Suarez, Rances Blanco, Teresita Badia, Dianelys Rivero, Alejandro Lopez-Requena, Armando Lopez and Enrique Montero

Department of Experimental Immunotherapy, Center of Molecular Immunology, 216 St., P.O. Box 16040, Havana 11600, Cuba

Key words: cytotoxicity, epidermal-growth-factor receptor (EGFR), flow cytometry, melanoma, overexpression, proliferation.

Abbreviations used: CDC, complement-dependent cytotoxicity; DMEM, Dulbecco's modified Eagle's medium; EGF, epidermal growth factor; EGFR, EGF receptor; FCS, foetal calf serum; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; RT, reverse transcriptase.

¹To whom correspondence should be addressed (email arlhee@cim.sld.cu).

EGFR [EGF (epidermal growth factor) receptor] overexpression correlates with poor prognosis and bad outcomes in different tumours. However, evidence for EGFR contribution in melanoma immunobiology is limited. We have expressed the full-length human EGFR gene in a murine melanoma cell line. EGFR protein expression in stably trnasfected B16 cells in culture was defined by immunoblotting, immunohistochemistry and FACS. Additionally, transfected cells became sensitive to the lysis induced with an anti-EGFR monoclonal antibody in the presence of complement. Exogenous human EGF addition induced cell proliferation, validating the transfected receptor functionality. Thus we have developed a system to express a functional EGFR in order to evaluate the potential contribution of EGFR expression in melanoma biology and its resulting relevance as a target for immunointerventions in nonepithelial tumours.

Received 8 January 2007/16 April 2007; accepted 2 May 2007

Published as Immediate Publication 2 May 2007, doi:10.1042/BA20070009

© 2007 Portland Press Ltd