Analysis of non-homologous end joining repair of DNA damage induced by high LET heavy ions

Wada, Seiichi; Kobayashi, Yasuhiko; Funayama, Tomoo; Sakashita, Tetsuya; Yokota, Yuichiro; Matsumoto, Yoshihisa*; Oto, Takayo*; Hosoi, Yoshio*; Suzuki, Norio*; Hamada, Nobuyuki*; Kakizaki, Takehiko*

Clustered DNA damages induced by high LET heavy ions are thought non-repairable or difficult to repair. However, much less is known about the reparability of clustered DNA damages. The major DNA repair pathway in mammalian cells is non-homologous end joining (NHEJ). In this study we investigated NHEJ pathway by exposure to high LET heavy ions, by analyzing responses of Ku to DNA damage induced by high LET heavy ions. pGFP and pGFP-Ku80 were transfected into xrs-5 cells (mutated in Ku80). Cells were irradiated with rays and ion beams (LET=2.7-1610 keV/m) at TIARA JAERI-Takasaki. Survival rates were measured by colony formation assay. To examine DNA damage and NHEJ pathway induced by heavy ion beams, H2AX and GFP signal on the nuclei were observed. Xrs5-GFP-Ku80 cells were radioresistant to rays and all ion beams, in comparison with xrs5-GFP cells. When the inactivation cross section was calculated to evaluate lethal effects per one particle, the difference of inactivation cross section between xrs5-GFP cells and xrs5-GFP-Ku80 cells became smaller with increasing LET. GFP and H2AX foci on the nuclei co-localized from 10 to 30 min after C (108 keV/m) and Ne ion (321 keV/m) irradiation. While GFP foci were observed for 10 min after Ar ion (1610 keV/m) irradiation, were not observed for 20 min. The difference of LET of ion beams influenced NHEJ responses.