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Mechanisms of resistance to erbitux (anti-epidermal growth factor receptor) combination therapy in pancreatic adenocarcinoma cells

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Journal of Gastrointestinal Surgery Aims and scope

Abstract

We previously demonstrated that pancreatic adenocarcinoma BxPC-3 xenografts display resistance to treatment with Erbitux, gemcitabine, and radiation, whereas MIA PaCa-2 xenografts are highly sensitive to the same therapy. Here, we elucidate in vitro mechanisms that may explain the observed differential response of epidermal growth factor receptor (EGFR) expressing pancreatic adenocarcinoma xenografts to Erbitux-based combination therapy in vivo. MIA PaCa-2 and BxPC-3 protein lysates were probed with antibodies to EGFR, ErbB2, ErbB3, and ErbB4. Constitutive ErbB3 activity was visualized by immunoblot analysis using anti-phosphotyrosine antibodies and receptor-specific immunoprecipitates. erbB2 and erbB3 gene expression in both cell lines was quantified with real-time polymerase chain reaction. Erbitux-induced internalization of EGFR was determined by flow cytometry following Erbitux treatment for different incubation times at 0°C and 37°C. MIA PaCa-2 and BxPC-3 protein extracts were also probed with anti-phospho-mitogen-activated protein kinase antibody after stimulation with EGF and in the presence of Erbitux. Although both cell lines expressed EGFR and ErbB2 protein, ErbB3 protein was selectively expressed by BxPC-3 cells, where it also showed evidence of constitutive phosphorylation. There was a 10-fold increase of erbB3 transcript levels in BxPC-3 cells compared with MIA PaCa-2. ErbB4 protein was not detectable in either cell line. Erbitux mediated EGFR internalization in MIA PaCa-2 cells after 2 hours of incubation, whereas it did not promote EGFR internalization in BxPC-3 cells. Likewise, EGF-dependent phosphorylation of MAPK p44/42 was blocked by Erbitux treatment in MIAPaCa-2 but not BxPC-3 cells. Erbitux selectively interfered with EGF-induced MAPK activation in MIA PaCa-2 but not BxPC-3 cells. Persistent MAPK activation and impaired in vitro internalization of EGFR by BxPC-3 pancreatic cancer cells may be due to constitutive ErbB3 signaling, facilitated by heterodimerization with EGFR, which may explain resistance to Erbitux-based combination therapy in vivo.

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Authors and Affiliations

  1. Department of Surgery, University of Alabama at Birmingham, Birmingham, Alabama

    J. Pablo Arnoletti M.D., Zhi-qiang Huang M.D., Ashley E. Hawkins M.D. & Selwyn M. Vickers M.D.

  2. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, Alabama

    Donald J. Buchsbaum Ph.D.

  3. Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama

    Muhamad B. Khazaeli Ph.D. & Matthias H Kraus M.D.

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  1. J. Pablo Arnoletti M.D.
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  2. Donald J. Buchsbaum Ph.D.
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  3. Zhi-qiang Huang M.D.
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  5. Muhamad B. Khazaeli Ph.D.
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  6. Matthias H Kraus M.D.
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  7. Selwyn M. Vickers M.D.
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Correspondence to Selwyn M. Vickers M.D..

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Arnoletti, J.P., Buchsbaum, D.J., Huang, Zq. et al. Mechanisms of resistance to erbitux (anti-epidermal growth factor receptor) combination therapy in pancreatic adenocarcinoma cells. J Gastrointest Surg 8, 960–970 (2004). https://doi.org/10.1016/j.gassur.2004.09.021

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  • DOI: https://doi.org/10.1016/j.gassur.2004.09.021

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