Effect of cetuximab and fractionated irradiation on tumour micro-environment

Santiago, Alina; Eicheler, Wolfgang; Bussink, Jan; Rijken, Paul; Yaromina, Ala; Beuthien-Baumann, Bettina; van der Kogel, Albert J.; Baumann, Michael; Krause, Mechthild

doi:10.1016/j.radonc.2010.07.007

« Previous Next »Radiotherapy & Oncology
Volume 97, Issue 2 , Pages 322-329, November 2010

Effect of cetuximab and fractionated irradiation on tumour micro-environment

Alina Santiago
- Department of Radiation Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- OncoRay Center for Radiation Research in Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
Wolfgang Eicheler
- Department of Radiation Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- OncoRay Center for Radiation Research in Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- Experimental Center, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
Jan Bussink
- Department of Radiation Oncology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
Paul Rijken
- Department of Radiation Oncology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
Ala Yaromina
- Department of Radiation Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- OncoRay Center for Radiation Research in Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
Bettina Beuthien-Baumann
- Department of Nuclear Medicine, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
Albert J. van der Kogel
- Department of Radiation Oncology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
Michael Baumann
- Department of Radiation Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- OncoRay Center for Radiation Research in Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- Experimental Center, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
Mechthild Krause
- Department of Radiation Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- OncoRay Center for Radiation Research in Oncology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany
- Corresponding author. Address: Department of Radiation Oncology, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstr. 74, 01307 Dresden, Germany.

Received 7 February 2010; received in revised form 4 July 2010; accepted 7 July 2010. published online 28 July 2010.

Abstract

Background and purpose

Previous experiments have shown that application of the anti-EGFR monoclonal antibody C225 (cetuximab) improves local tumour control after irradiation in FaDu human squamous cell carcinoma (hSCC) due to the combined effect of decreased repopulation and improved reoxygenation. The present study investigates early changes of the pimonidazole hypoxic fraction of FaDu tumours and the expression and phosphorylation of the EGFR and its downstream signal transduction molecules after treatment with C225 alone or in combination with irradiation.

Material and methods

FaDu tumour xenografts were irradiated with up to 3×3Gy with or without additional C225 treatment and excised at different time points. Tumour hypoxia was evaluated using pimonidazole. EGFR expression and phosphorylation and intratumoural distribution of C225 were assessed by immunofluorescence analysis. Western blots were performed to evaluate expression and phosphorylation of EGFR, ErbB2, AKT and MAPK (ERK1/2).

Results

Hypoxia did not change during the 4days of treatment in the tumours treated with C225 alone or combined with irradiation. C225 treatment led to downregulation of the total EGFR in FaDu tumours, accompanied by a change of the spatial distribution of the receptor favouring the membranous expression. An induction of phosphorylation of the EGFR (tyr992, tyr1173) was observed with C225 alone or combined with irradiation. AKT phosphorylation was decreased, whereas MAPK phosphorylation remained unchanged. C225 membrane staining was homogeneously distributed over the whole tumour with no differences between hypoxic and non-hypoxic tumour cells.

Conclusion

Pimonidazole-hypoxia of FaDu tumours during the initial part of fractionated irradiation is not influenced by C225, indicating that external hypoxia markers may not be promising as biomarkers for tumour response to combined treatment. The downregulation of the total EGFR, but at the same time higher membrane staining, as well as the changes in downstream signal transduction molecules, warrants further investigation in other tumour models.

Keywords: EGFR inhibition, Radiotherapy, Cetuximab, C225, Monoclonal antibodies, Hypoxia, Pimonidazole, Tumour micro-environment, Biomarkers, Signal transduction