Flow cytometric assessment of DNA double-strand break and repair kinetics in prediction of intrinsic radiosensitivity

Objective(s): To enhance the efficiency of radiotherapy (RT), implementation of individual-based treatment is essential. In this way, determining individual intrinsic radiosensitivity (IRS) can be useful to achieve minimal adverse effects of RT. The present study aimed to identify IRS of breast cancer (BC) patients through determination of radiation-induced DNA double-strand breaks (DSBs), repair kinetics, and acute normal tissue complications induced by RT.Materials and Methods: DSBs induction and its repair kinetics in ۵۰ BC patients’ lymphocytes were analyzed by flow cytometric analysis of H۲AX Ser-۱۳۹ phosphorylation at ۳۰ min, ۳ and ۲۴ hr after in vitro irradiation. In vivo skin dosimetry was done by GAFChromic films and acute skin toxicity was scored by radiation oncologists according to the criteria of Radiation Therapy and Oncology Group (RTOG) in all patients with similar prescribed treatment.Results: The average surface dose for patients ranged from ۰.۹۲ to ۱.۹ Gy and correlation analysis showed no significant relationship with weekly acute skin reactions. Formation of γH۲AX after ۳۰ min, slope of dose-response curve and repair kinetics of DSBs after ۳ and ۲۴ hr (intrinsic radiosensitivity) were significantly correlated with the RTOG scores following irradiation (clinical radiosensitivity) (r=۰.۴۸ and P-value<۰.۰۰۰۱, r=۰.۷۲ and P-value<۰.۰۰۰۱, r=۰.۴۸ and P-value<۰.۰۰۱, and finally r=۰.۵۳ and P-value<۰.۰۰۱, respectively; (using Pearson’s correlation test). Conclusion: Flow cytometric analysis of DNA DSBs by γH۲AX measurement has the potential to be developed into a clinical predictor for identifying the overreactor patients prior to RT. Our result suggests that the slope-related quantity based on the linear pattern of the dose-response curve has the merit to predict overreactor patients with a sensitivity of ۸۹% and a specificity of ۹۴%.

Keywords:

Breast Cancer , DNA damage , Double-Strand Break , Radiosensitivity , Radiotherapy