Inhibition of radiation-induced EGFR nuclear import by C225 (Cetuximab) suppresses DNA-PK activity

• Published in: Radiotherapy and oncology 2005-08, Vol.76 (2), p.157-161
• Main Authors: Dittmann, Klaus, Mayer, Claus, Rodemann, Hans-Peter
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal - pharmacology; Antibodies, Monoclonal, Humanized; Antineoplastic Agents - pharmacology; C225; Cell Line, Tumor - metabolism; Cell Line, Tumor - radiation effects; Cell Nucleus - metabolism; Cetuximab; Cytoplasm - metabolism; DNA Repair; DNA-Activated Protein Kinase - metabolism; DNA-PK; EGFR; Enzyme Activation; Humans; Radiation; Radiation Tolerance; Receptor, Epidermal Growth Factor - antagonists & inhibitors; Receptor, Epidermal Growth Factor - metabolism; Receptor, Epidermal Growth Factor - radiation effects; Tumor Cells, Cultured
• ISSN: 0167-8140; 1879-0887
• DOI: 10.1016/j.radonc.2005.06.022

Inhibition of EGFR-function can induce radiosensitization in tumor cells. Purpose of our investigation was to identify the possible molecular mechanism of radiosensitization following treatment with anti-EGFR-antibody C225 (Cetuximab). The effect of C225 on radiation response was determined in human cell lines of bronchial carcinoma (A549) and breast adenoma cells (MDA MB 231). The molecular effects of C225 on EGFR-function after irradiation were analyzed applying western blotting, immune-precipitation and kinase assays. Effects on DNA-repair were detected by quantification of γ-H2AX positive foci 24 h after irradiation. The EGFR specific antibody C225 induced radiosensitization in A549 and also in MDA MB 231 cells. Radiosensitization in A549 was associated with blockage of radiation-induced EGFR transport into the nucleus, and immobilized the complex of EGFR with DNA-dependent protein kinase (DNA-PK) in the cytoplasm. As a consequence radiation-induced DNA-PK activation was abolished, a process that is essential for DNA-repair after radiation exposure. Likewise C225 treatment increased the residual amount of γ-H2AX-positive foci 24 h after irradiation in A549 and in MDA MB 231 cells. Our results suggest that irradiation induced DNA-PK activation—essential for DNA repair—may be hampered specifically by use of the anti-EGFR-antibody C225. This process is associated with radiosensitization.