Wang, H., Wang, X., Iliakis, G. and Wang, Y. Caffeine Could Not Efficiently Sensitize Homologous Recombination Repair-Deficient Cells to Ionizing Radiation-Induced Killing. Radiat. Res. 159, 420–425 (2003).

Caffeine inhibits ATM and ATR, two important checkpoint regulators, abolishes ionizing radiation-induced checkpoint response, and radiosensitizes cells. Radiation-induced DNA double-strand breaks (DSBs) are repaired by two major processes, homologous recombination repair (HRR) and nonhomologous end joining (NHEJ). It remains unclear which repair process, HRR or NHEJ, is affected when the checkpoint responses are abolished by caffeine. In this study we observed the effect of caffeine on gene-targeted DT40 chicken lymphoblast cells. We show that caffeine efficiently abolishes S- and G₂-phase checkpoint responses after irradiation in all cell lines tested and greatly radiosensitizes wild-type and ATM^–/– cells, the partially checkpoint-deficient cells. However, caffeine has a much smaller radiosensitizing effect on RAD54^–/– cells and has no effect on RAD51-deficient cells. RAD51 and RAD54 are the important factors for HRR. Our results indicate that the checkpoint responses abolished by caffeine (S and G₂) mainly affect HRR, which results in cell radiosensitization.