For X-irradiated normal human fibroblasts, only half of cell inactivation results from chromosomal damage.

摘要

PURPOSE: To study the relationship between residual double-strand breaks (dsbs), chromosomal damage, and cell inactivation for X-irradiated normal human fibroblasts. METHODS AND MATERIALS: The experiments were performed with 12 normal human fibroblast strains and, for comparison, a fibroblast line from a LiFraumeni patient (LFS2800), a squamous cell carcinoma line (FaDu), and CHO cells. Cells were irradiated in plateau phase, which was followed by immediate or delayed (14 h) plating. Chromosomal damage was measured by metaphase technique and loss of proliferative capacity by colony-forming assay. The data obtained were compared with residual double-strand breaks measured previously (Dikomey et al. IJROBP 2000;46:481-490). RESULTS: For each fibroblast strain, the number of lethal chromosome aberrations (CAs) increased with dose, but with a substantial variation among the strains (coefficient of variation = 20%-26%). The number of lethal aberrations was significantly correlated with the number of residual dsbs measured for the same strain (r(2) = 0.71, p = 0.0006). The residual dsbs were assumed to represent both non- and also mis-rejoined dsbs. There was a significant correlation between lethal aberrations and cell survival, but only for delayed and not immediate plating (r(2) = 0.69, p < 0.0008 vs. r(2) = 0.19, p = 0.16). For delayed plating, the ratio between lethal events (LEs) and CAs amounted to LE:CA = 2.0 +/- 0.05:1, indicating that on average, only half of cell inactivation resulted from chromosomal damage. The other 50% was attributed to the p53-dependent permanent G1 arrest, because cells lacking in functional p53 (LFS2800, FaDu, CHO) showed a ratio of LE:CA = 1.01 +/- 0.02:1. CONCLUSION: On average, up to 50% of the inactivation of X-irradiated normal human fibroblasts is a result of lethal chromosome aberrations, whereas the rest is due to a p53-dependent process, probably permanent G1 arrest.