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Expression and activity of EGFR in human cutaneous melanoma cell lines and influence of vemurafenib on the EGFR pathway

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Abstract

Data regarding the expression of epidermal growth factor receptor (EGFR) in melanoma and its role in the tumor biology are conflicting. In BRAF V600-mutant melanomas, the expression of EGFR has been associated with acquired resistance to BRAF inhibitors. In this study, we assessed EGFR expression and downstream signaling activity in a panel of melanoma cell lines and we investigated the effects of the BRAF inhibitor vemurafenib on expression of EGFR and its downstream effectors in a subgroup of BRAF-mutant melanoma cells. Three out of 10 melanoma cell lines expressed EGFR. Downstream signaling via ERK and AKT was responsive to either stimulation by EGF or inhibition by erlotinib. Constitutive activation of ERK occurred in all the cell lines investigated whereas constitutive activation of AKT only in three cell lines. Constitutive activation of ERK and AKT was independent from EGFR expression. Vemurafenib did not affect EGFR expression in general, but it increased EGFR phosphorylation in the cell line SkMel5. Induced EGFR phosphorylation was sensitive to treatment with erlotinib. Vemurafenib efficiently blocked ERK activation in all the BRAF-mutant cell lines tested, whereas its effects on AKT activation were dissimilar in the different cell lines. Our data suggest that EGFR is functional but usually inactive in EGFR high-expressing cell lines. Basal EGFR expression unlikely represents a biomarker for predicting the sensitivity to vemurafenib in melanoma, but EGFR activation might represent a mechanism of vemurafenib resistance in a subset of melanoma cells.

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Conflicts of Interest

All authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Hematology and Medical Oncology, Charité, Campus Benjamin Franklin, Hindenburgdamm 30, 12200, Berlin, Germany

    Alexander Gross, Annett Niemetz-Rahn, Anika Nonnenmacher, Johannes Tucholski, Ulrich Keilholz & Alberto Fusi

  2. Department of Medical Oncology, The Christie NHS Foundation Trust, Manchester, M20 4BX, UK

    Alberto Fusi

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  1. Alexander Gross
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  2. Annett Niemetz-Rahn
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Correspondence to Alexander Gross.

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Fig. S1

Cellular localization of EGFR. Five human melanoma cell lines were stained with a fluorescent antibody against EGFR. To evaluate extracellular EGFR expression, cells were stained directly after scratching and washing. For intracellular staining, cells were fixed and permeabilized prior to incubation with the fluorescent antibody. The human squamous cell carcinoma cell line FaDu served as positive control (GIF 59 kb)

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Gross, A., Niemetz-Rahn, A., Nonnenmacher, A. et al. Expression and activity of EGFR in human cutaneous melanoma cell lines and influence of vemurafenib on the EGFR pathway. Targ Oncol 10, 77–84 (2015). https://doi.org/10.1007/s11523-014-0318-9

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  • DOI: https://doi.org/10.1007/s11523-014-0318-9

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