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Afatinib, an Irreversible EGFR Family Inhibitor, Shows Activity Toward Pancreatic Cancer Cells, Alone and in Combination with Radiotherapy, Independent of KRAS Status

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Abstract

Background

Pancreatic adenocarcinoma is characterized by a high frequency of KRAS mutations and frequent deregulation of the epidermal growth factor receptor (EGFR) and other EGFR family members such as HER2/ErbB2. The EGFR inhibitor erlotinib is approved for treatment of pancreatic cancer, but has shown modest activity in most patients.

Objective

Here we investigated the activity of afatinib, a second-generation irreversible pan-EGFR family kinase inhibitor, alone or in combination with ionizing radiation, toward pancreatic cancer cells.

Methods

The influence of afatinib on cell proliferation, cell cycle distribution, clonogenic survival, nuclear fragmentation, ploidy, and centrosome amplification following irradiation was determined. Expression and phosphorylation of HER receptors, Akt, DNA-PKcs, and ERK1/2 was characterized by Western blot analysis.

Results

Afatinib was growth-inhibitory for all three cell lines but cytotoxic only toward BxPC3 (KRAS wt) and Capan-2 (KRAS mut) cells, both of which express high levels of EGFR, HER2, and HER3 receptors. Afatinib increased the radiosensitivity of BxPC3 and Capan-2 cells, prevented the radio-induced phosphorylation of Akt, and induced mitotic catastrophe following irradiation. In comparison, Panc-1 cells (KRAS mut) expressing low levels of EGFR family receptors were resistant to afatinib-induced radiosensitization.

Limitations

These results must be confirmed in vivo.

Conclusions

Afatinib showed cytotoxic and radiosensitizing effects toward a subset of pancreatic cancer cells which was closely correlated with expression of EGFR, HER2, and HER3 receptors, but not with KRAS status.

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Acknowledgments

The authors wish to thank Valérie Fauquette (INSERM U837, Lille) for the generous gift of Capan-2 cells and Janet Hall (Institut Curie, Orsay) for helpful discussions.

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Author notes

    Authors and Affiliations

    1. Institut Curie, Centre de Recherche, 91405, Orsay, France

      Florence Huguet, Marie Fernet, Nicole Giocanti & Vincent Favaudon

    2. Institut National de la Santé et de la Recherche Médicale (INSERM) U612, 91405, Orsay, France

      Florence Huguet, Marie Fernet, Nicole Giocanti & Vincent Favaudon

    3. Institut Universitaire de Cancérologie, Université Pierre et Marie Curie, Sorbonne Universités, 75005, Paris, France

      Florence Huguet & Annette K. Larsen

    4. Assistance Publique – Hôpitaux de Paris, Hôpitaux Universitaires Est Parisien, Hôpital Tenon, Service d’Oncologie Radiothérapie, 75020, Paris, France

      Florence Huguet

    5. Laboratory of Cancer Biology and Therapeutics, Centre de Recherche Saint-Antoine, 75012, Paris, France

      Florence Huguet & Annette K. Larsen

    6. Institut National de la Santé et de la Recherche Médicale (INSERM) U938, 75012, Paris, France

      Florence Huguet & Annette K. Larsen

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    1. Florence Huguet
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    Correspondence to Florence Huguet.

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    F. Huguet, M. Fernet, N. Giocanti, V. Favaudon, and A.K. Larsen declare no conflict of interest.

    F. Huguet was supported in part by the Nuovo-Soldati Research Foundation.

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    Vincent Favaudon and Annette K. Larsen contributed equally to this work.

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    Huguet, F., Fernet, M., Giocanti, N. et al. Afatinib, an Irreversible EGFR Family Inhibitor, Shows Activity Toward Pancreatic Cancer Cells, Alone and in Combination with Radiotherapy, Independent of KRAS Status. Targ Oncol 11, 371–381 (2016). https://doi.org/10.1007/s11523-015-0403-8

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    • DOI: https://doi.org/10.1007/s11523-015-0403-8

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