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Functional expression of human-epidermal-growth-factor receptor in a melanoma cell line
Arlhee Diaz1, Eduardo Suarez, Rances Blanco
1Department of Experimental Immunotherapy, Center of Molecular Immunology, Havana, Cuba. arlhee@cim.sld.cu
Abstract:
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.
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