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Epidermal growth factor receptor status and persistent activation of Akt and p44/42 MAPK pathways correlate with the effect of cetuximab in head and neck and colon cancer cell lines

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Abstract

Objective

The aim of this study was to investigate the effect of epidermal growth factor receptor (EGFR) blockade on cell survival and on downstream signalling pathways using the monoclonal antibody cetuximab.

Methods

We used three colon cancer cell lines, of which one was EGFR-negative, and two head and neck squamous cell carcinoma (HNSCC) lines. EGFR expression and gene copy number were measured by immunohistochemistry and FISH analysis, respectively. The effect of cetuximab, irradiation or the combination of both on cell growth was estimated by SRB assay. Western blotting was used to determine the phosphorylation of intracellular signalling proteins and cell cycle phase distribution was measured by flow cytometry.

Results

The addition of cetuximab had only limited effects on cell growth, with a maximum inhibition of approximately 30%, but was correlated with the amount of protein expression and gene copy number of EGFR. When combined with irradiation, the effect of cetuximab was only additive and not dependent on the inherent radio-sensitivity of the cell lines. Persistent phosphorylation of Akt and/or p44/42 MAPK was detected by western blot in all of the cell lines, whereas there was no phosphorylation of Jak2 or STAT3.

Conclusions

None of these factors alone could predict the sensitivity to cetuximab. Rather, the results suggest that it might be necessary to determine the activation status of several intracellular signalling proteins to better predict the sensitivity to cetuximab treatment.

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Acknowledgments

We thank Christina Boll for expert technical assistance with cell cultures and immunohistochemistry. This work was supported by the Lund University Hospital Bequest Fund, the Swedish Cancer Society (1304-B06-20XCC and 4839-B05-03PCC), the King Gustaf V Jubilee Fund (05-4161), government funding of clinical research in Swedish healthcare, Scania Region R & D funding, and the Laryngfonden.

Conflict of interest statement

I certify that there is no actual or potential conflict of interest in relation to this article.

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Authors and Affiliations

  1. Department of Otorhinolaryngology/Head and Neck Surgery, Lund University Hospital, 221 85, Lund, Sweden

    Takashi Yamatodani & Johan Wennerberg

  2. Department of Oncology, Lund University Hospital, 221 85, Lund, Sweden

    Lars Ekblad, Elisabeth Kjellén & Anders Johnsson

  3. Department of Otorhinolaryngology/Head and Neck Surgery, Hamamatsu University School of Medicine, Hamamatsu, 431-3192, Japan

    Takashi Yamatodani & Hiroyuki Mineta

Authors
  1. Takashi Yamatodani
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  2. Lars Ekblad
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  3. Elisabeth Kjellén
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  4. Anders Johnsson
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  5. Hiroyuki Mineta
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  6. Johan Wennerberg
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Correspondence to Lars Ekblad.

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Yamatodani, T., Ekblad, L., Kjellén, E. et al. Epidermal growth factor receptor status and persistent activation of Akt and p44/42 MAPK pathways correlate with the effect of cetuximab in head and neck and colon cancer cell lines. J Cancer Res Clin Oncol 135, 395–402 (2009). https://doi.org/10.1007/s00432-008-0475-2

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  • DOI: https://doi.org/10.1007/s00432-008-0475-2

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