Epidermal growth factorIn tumor cells regulation of DNA double strand break repair through EGF receptor involves both NHEJ and HR and is independent of p53 and K-Ras status
Section snippets
Cell lines
Bronchial carcinoma cells A549 and H1299, cervix carcinoma cells HeLa and vervet monkey fibroblasts CV1 were grown in D-MEM (Gibco) containing 10% FBS (Biochrom AG) and 4 mM glutamine (Gibco) at 37 °C, 10% CO2 and 100% humidification.
X-irradiation
Cells were irradiated at room temperature with 200 kV X-rays (Gulmay RS225, Gulmay Medical Ltd., 15 mA, 0.8 mm Be + 0.5 mm Cu filtering; dose rate of 1.2 Gy/min).
Ligands and inhibitors
EGF, ARG, TGFα and DMSO, Sigma; cetuximab (Erbitux®), Merck; erlotinib (Tarceva®), Roche.
Western blot
Proteins from whole
Modulation of DSB repair by different EGFR ligands
We have recently shown that the stimulation of EGFR by EGF activates the overall DSB repair and especially NHEJ [16]. We now tested whether this effect can be observed when EGFR is stimulated by other specific ligands such as amphiregulin (ARG) and transforming growth factor α (TGFα). Treatment of A549 cells with TGFα or ARG was found to result in a concentration-dependent phosphorylation of EGFR and ERK1/2, whereby the strongest activation was measured for EGF and the weakest for ARG (Fig. 1
Discussion
The aim of this study was to elucidate whether the regulation of DSB repair via EGFR is a common phenomenon or restricted to specific conditions. This is important not only to understand the underlying mechanisms of this regulation but also to determine whether EGFR can be used as a general target or only in some specific cell lines.
In contrast to previous studies focussing on EGF-mediated EGFR activation [14], [16], it was shown here for the first time that both overall DSB repair and NHEJ are
Conflict of interest statement
It is hereby confirmed that all authors disclose all financial and personal relationships with other people or organisations that could inappropriately influence (bias) their work.
Acknowledgments
The authors greatly acknowledge the technical assistance of K. Hoffer and B. Riepen. This project was supported by Deutsche Forschungsgemeinschaft (DFG PAK 190, DI 457/8-1; U.K.P., J.D.-D., E.D.) and Deutsche Krebshilfe (Grant No. 107889; T.R., J.D.-D.).
References (27)
Molecular radiation biology: perspectives for radiation oncology
Radiother Oncol
(2009)- et al.
EGFR-targeted anti-cancer drugs in radiotherapy: preclinical evaluation of mechanisms
Radiother Oncol
(2007) - et al.
Radiotherapy plus cetuximab for locoregionally advanced head and neck cancer: 5-year survival data from a phase 3 randomised trial, and relation between cetuximab-induced rash and survival
Lancet Oncol
(2010) - et al.
Receptor signaling as a regulatory mechanism of DNA repair
Radiother Oncol
(2009) - et al.
Mechanisms of human DNA repair: an update
Toxicology
(2003) - et al.
Radiation-induced epidermal growth factor receptor nuclear import is linked to activation of DNA-dependent protein kinase
J Biol Chem
(2005) - et al.
The epidermal growth factor receptor modulates DNA double-strand break repair by regulating non-homologous end-joining
DNA Repair
(2010) - et al.
Inhibition of radiation-induced EGFR nuclear import by C225 (Cetuximab) suppresses DNA-PK activity
Radiother Oncol
(2005) - et al.
Radioresistance of K-Ras mutated human tumor cells is mediated through EGFR-dependent activation of PI3K-AKT pathway
Radiother Oncol
(2005) - et al.
Impact of epidermal growth factor receptor expression on survival and pattern of relapse in patients with advanced head and neck carcinoma
Cancer Res
(2002)
Radiosensitivity of tumor cell lines after pretreatment with the EGFR tyrosine kinase inhibitor ZD1839 (Iressa)
Strahlenther Onkol
Epidermal growth factor receptor blockade with C225 modulates proliferation, apoptosis, and radiosensitivity in squamous cell carcinomas of the head and neck
Cancer Res
Blockage of epidermal growth factor receptor-phosphatidylinositol 3-kinase-AKT signaling increases radiosensitivity of K-RAS mutated human tumor cells in vitro by affecting DNA repair
Clin Cancer Res
Cited by (53)
Relationships between DNA repair and RTK-mediated signaling pathways
2021, Biochimica et Biophysica Acta - Reviews on CancerModulating the DNA Damage Response to Improve Treatment Response in Cervical Cancer
2017, Clinical OncologyCytotoxicity and genotoxicity effects of arsenic trioxide on SQ20B human laryngeal carcinoma cells
2017, Experimental and Toxicologic PathologyCitation Excerpt :Several reports have involved the implication of the EGFR pathway in the regulation of DNA DSB repair by positive downstream of the homologous recombination as well as the non homologous end-joining (Li et al., 2008; Myllynen et al., 2011). Thus, the activation of EGFR decreases the number of residual DNA DSBs, whereas the number of γ-H2AX-positive DSB foci was clearly increased when EGFR was blocked in other lung cancer cell lines (Myllynen et al., 2011). Herbert and Snow (2012) showed that the H2AX deficiency could be responsible for the chromosomal instability as well as cancer due to the alteration of this DNA reparation process.
Radiation DNA damage and use in cancer/therapeutics-translation of radiation modifiers
2016, DNA Repair in Cancer Therapy: Molecular Targets and Clinical Applications: Second EditionPhosphatidylinositol 3-kinase/Akt signaling as a key mediator of tumor cell responsiveness to radiation
2015, Seminars in Cancer BiologyCitation Excerpt :The status of targetability of the PI3K/Akt pathway in hematological malignancies has also been well reviewed by other authors [160–162]. Previous reports showed a direct EGFR-Akt correlation, indicating that EGFR was a major regulator of Akt-dependent DNA-DSB repair [97,163–165] after irradiation. Moreover, the impact of Akt activity on DNA-DSB repair and radioresistance in tumor cells from different origins has also been demonstrated [23,102,128,166,167].