Ludwików G, Hofer K G, Bao S P, Ludwików F
Department of Radiobiology, Swedish University of Agricultural Sciences, Uppsala, Sweden.
Int J Radiat Biol. 1996 Aug;70(2):177-87. doi: 10.1080/095530096145175.
Chinese hamster ovary (CHO) cells were synchronized in M phase by mitotic selection, and then re-synchronized with aphidicolin at the G1/S phase border. The cells were labelled in early-S phase by 10 min exposure to 125I-iododeoxyuridine and then cultured (chased) in non-radioactive medium for 0.5, 3 or 5h, followed by harvesting and freezing to accumulate the desired number of 125I decays. Cell damage was assessed by evaluating colony formation, micronucleus formation and chromosome aberrations. These biological estimators of damage showed that the cytocidal effect of 125I decay increased with the duration of the post-labelling chase period: the highest level of damage was found in cells from the 5 h chase period and the lowest in the cells from the 0.5 h chase period. Survival curves for the three chase periods displayed low-dose hyper-radiosensitivity for 0 to 20 125I decays cell-1. The results indicate that the repair of DNA double-strand breaks (DSBs) may depend on the maturation stage of chromatin and an explanation of this finding is proposed which invokes the homologous recombination model for DSB repair.
中国仓鼠卵巢(CHO)细胞通过有丝分裂选择同步于M期,然后在G1/S期边界用阿非迪霉素重新同步。细胞在早S期通过暴露于125I-碘脱氧尿苷10分钟进行标记,然后在无放射性培养基中培养(追踪)0.5、3或5小时,随后收获并冷冻以积累所需数量的125I衰变。通过评估集落形成、微核形成和染色体畸变来评估细胞损伤。这些损伤的生物学指标表明,125I衰变的杀细胞效应随着标记后追踪期的延长而增加:在5小时追踪期的细胞中发现的损伤水平最高,在0.5小时追踪期的细胞中最低。三个追踪期的存活曲线在0至20次125I衰变/细胞时显示出低剂量超放射敏感性。结果表明,DNA双链断裂(DSB)的修复可能取决于染色质的成熟阶段,并提出了对此发现的一种解释,该解释援引了DSB修复的同源重组模型。