Misrepair of DNA double-strand breaks after exposure to heavy-ion beams causes a peak in the LET-RBE relationship with respect to cell killing in DT40 cells

Abstract

To determine the radiobiological mechanisms underlying relative biological effectiveness (RBE) and the repair efficiencies of DNA double-strand breaks (DSBs) as a function of linear energy transfer (LET), we exposed cells of the chicken B-lymphocyte cell line DT40 and its DSB repair pathway-deficient derivatives to heavy-ion beams produced at the Heavy-Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Sciences (NIRS), Chiba, Japan. The relationship between LET and cell lethality was investigated in the DNA DSB repair gene knockouts Ku70(-/-), Rad54(-/-), and Ku70(-/-)Rad54(-/-), and in the wild-type cells. We found that cell-cycle stage and activity of the DNA DSB repair pathways influence LET-mediated biological effects. An expected LET-RBE relationship was observed in the cells capable of DNA repair, but no peak was found in the RBE with respect to cell survival in the Ku70(-/-)Rad54(-/-) cells or in Ku70(-/-) cells in the G1 and early S cell-cycle phases (when no sister chromatids were present and homologous recombination could not occur). These findings suggest that the peak in RBE is caused by deficient repair of the DNA DSBs.

Keywords: heavy ion; linear energy transfer; misrepair; relative biological effectiveness; sensitivity.

Fig. 1.

Survival curves for the (a) DT40 wild-type, (b) Rad54^−/−, (c) Ku70^−/−Rad54^−/−, and (d) Ku70^−/− cells in different cell-cycle phases exposed to X-rays or Ne, Si, Ar or Fe ions with different LETs. The cell strains, ions and the LETs (keV/µm) are indicated.

Fig. 2.

LET versus D₁₀ values for cells in different knockout status (wild-type, Rad54^−/−, Ku70^−/−Rad54^−/− or Ku70^−/− ). Ku70^−/− cells at different cell cycle stages were divided into two subtypes: Ku70^−/−(f) (in the G₁/early-S phase), or Ku70^−/−(s) (in the late-S/G₂ phase). Plots of Ku70^−/−Rad54^−/− and Ku70^−/−(f) are fitted together. Each mark corresponds to inversed D₁₀ values from a single experiment.

Fig. 3.

Dose–response for the frequency of chromosome #1 fragmentation in the wild-type and Ku70^−/−Rad54^−/− cells after exposure to X-rays, Ar ions or Fe ions with different LETs (in keV/µm). Values with error bars are calculated from at least three experiments.

Fig. 4.

Induction yields of chromosome #1 fragmentation per Gy in wild-type or Ku70^−/−Rad54^−/− cells exposed to heavy ions, as a function of LET. Note that the vertical axis is inverted to be seen as the same as the D₁₀ spectrum in Fig. 2. The lines represent a simple smooth fit.