Impaired DNA Repair Fidelity in a Breast Cancer Patient With Adverse Reactions to Radiotherapy

Abstract

We tested the hypothesis that differences in DNA double-strand break (DSB) repair fidelity underlies differences in individual radiosensitivity and, consequently, normal tissue reactions to radiotherapy. Fibroblast cultures derived from a radio-sensitive (RS) breast cancer patient with grade 3 adverse reactions to radiotherapy were compared with normal control (NC) and hyper-radiosensitive ataxia-telangiectasia mutated (ATM) cells. DSB repair and repair fidelity were studied by Southern blotting and hybridization to Alu repetitive sequence and to a specific 3.2-Mbp NotI restriction fragment on chromosome 21, respectively. Results for DNA repair kinetics using the NotI fidelity assay showed significant differences (P < 0.001) with higher levels of misrepaired (misrejoined and unrejoined) DSBs in RS and ATM compared with NC. At 24-h postradiation, the relative fractions of misrepaired DSBs were 10.64, 23.08, and 44.70% for NC, RS, and ATM, respectively. The Alu assay showed significant (P < 0.05) differences in unrepaired DSBs only between the ATM and both NC and RS at the time points of 12 and 24 h. At 24 h, the relative percentages of DSBs unrepaired were 1.33, 3.43, and 12.13% for NC, RS, and ATM, respectively. The comparison between the two assays indicated an average of 5-fold higher fractions of misrepaired (NotI assay) than unrepaired (Alu assay) DSBs. In conclusion, this patient with increased radiotoxicity displayed more prominent misrepaired than unrepaired DSBs, suggesting that DNA repair fidelity is a potential marker for the adverse reactions to radiotherapy. More studies are required to confirm these results and further develop DSB repair fidelity as a hallmark biomarker for interindividual differences in radiosensitivity.

Keywords: Alu sequence; DNA double-strand breaks; NotI fragment; adverse reactions to radiotherapy; misrepair; pulsed field gel electrophoresis; radiosensitivity; repair fidelity.