BRCA1, FANCD2 and Chk1 are potential molecular targets for the modulation of a radiation-induced DNA damage response in bystander cells

TY - JOUR

T1 - BRCA1, FANCD2 and Chk1 are potential molecular targets for the modulation of a radiation-induced DNA damage response in bystander cells

AU - Burdak-Rothkamm, Susanne

AU - Rothkamm, Kai

AU - McClelland, Keeva

AU - Al Rashid, Shahnaz T

AU - Prise, Kevin M

N1 - Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

PY - 2015/1/28

Y1 - 2015/1/28

N2 - Radiotherapy is an important treatment option for many human cancers. Current research is investigating the use of molecular targeted drugs in order to improve responses to radiotherapy in various cancers. The cellular response to irradiation is driven by both direct DNA damage in the targeted cell and intercellular signalling leading to a broad range of bystander effects. This study aims to elucidate radiation-induced DNA damage response signalling in bystander cells and to identify potential molecular targets to modulate the radiation induced bystander response in a therapeutic setting. Stalled replication forks in T98G bystander cells were visualised via bromodeoxyuridine (BrdU) nuclear foci detection at sites of single stranded DNA. γH2AX co-localised with these BrdU foci. BRCA1 and FANCD2 foci formed in T98G bystander cells. Using ATR mutant F02-98 hTERT and ATM deficient GM05849 fibroblasts it could be shown that ATR but not ATM was required for the recruitment of FANCD2 to sites of replication associated DNA damage in bystander cells whereas BRCA1 bystander foci were ATM-dependent. Phospho-Chk1 foci formation was observed in T98G bystander cells. Clonogenic survival assays showed moderate radiosensitisation of directly irradiated cells by the Chk1 inhibitor UCN-01 but increased radioresistance of bystander cells. This study identifies BRCA1, FANCD2 and Chk1 as potential targets for the modulation of radiation response in bystander cells. It adds to our understanding of the key molecular events propagating out-of-field effects of radiation and provides a rationale for the development of novel molecular targeted drugs for radiotherapy optimisation.

AB - Radiotherapy is an important treatment option for many human cancers. Current research is investigating the use of molecular targeted drugs in order to improve responses to radiotherapy in various cancers. The cellular response to irradiation is driven by both direct DNA damage in the targeted cell and intercellular signalling leading to a broad range of bystander effects. This study aims to elucidate radiation-induced DNA damage response signalling in bystander cells and to identify potential molecular targets to modulate the radiation induced bystander response in a therapeutic setting. Stalled replication forks in T98G bystander cells were visualised via bromodeoxyuridine (BrdU) nuclear foci detection at sites of single stranded DNA. γH2AX co-localised with these BrdU foci. BRCA1 and FANCD2 foci formed in T98G bystander cells. Using ATR mutant F02-98 hTERT and ATM deficient GM05849 fibroblasts it could be shown that ATR but not ATM was required for the recruitment of FANCD2 to sites of replication associated DNA damage in bystander cells whereas BRCA1 bystander foci were ATM-dependent. Phospho-Chk1 foci formation was observed in T98G bystander cells. Clonogenic survival assays showed moderate radiosensitisation of directly irradiated cells by the Chk1 inhibitor UCN-01 but increased radioresistance of bystander cells. This study identifies BRCA1, FANCD2 and Chk1 as potential targets for the modulation of radiation response in bystander cells. It adds to our understanding of the key molecular events propagating out-of-field effects of radiation and provides a rationale for the development of novel molecular targeted drugs for radiotherapy optimisation.

KW - Ataxia Telangiectasia Mutated Proteins/genetics

KW - BRCA1 Protein/metabolism

KW - Blotting, Western

KW - Brain Neoplasms/genetics

KW - Bystander Effect/genetics

KW - Cell Proliferation/radiation effects

KW - Checkpoint Kinase 1

KW - DNA Damage/genetics

KW - DNA Repair/genetics

KW - DNA Replication/genetics

KW - Fanconi Anemia Complementation Group D2 Protein/metabolism

KW - Flow Cytometry

KW - Glioma/genetics

KW - Humans

KW - Immunoenzyme Techniques

KW - Mutation/genetics

KW - Phosphorylation/radiation effects

KW - Protein Kinases/metabolism

KW - Signal Transduction/radiation effects

KW - Tumor Cells, Cultured

KW - X-Rays

U2 - 10.1016/j.canlet.2014.09.043

DO - 10.1016/j.canlet.2014.09.043

M3 - SCORING: Journal article

C2 - 25304378

VL - 356

SP - 454

EP - 461

JO - CANCER LETT

JF - CANCER LETT

SN - 0304-3835

IS - 2 Pt B

ER -