Olive P L, Banáth J P
Medical Biophysics Department, British Columbia Cancer Research Centre, Vancouver, Canada.
Mutat Res. 1993 Oct;294(3):275-83. doi: 10.1016/0921-8777(93)90010-e.
Previous results using a variety of methods have shown no significant difference in induction or rejoining of radiation-induced DNA single-strand breaks (SSBs) as a function of cell-cycle position. However, concurrent analysis of DNA damage and cell-cycle position measured using the alkaline comet assay indicates cycle-dependent differences in "tail moment" which has been shown to be a measure of SSBs. Unirradiated S phase cells showed a significantly higher tail moment presumably as a result of the presence of active replication sites. The time required to repair half of the damage appeared to be consistently shorter for G1 cells than for cells in other phases of the cell cycle (3.7 min versus approximately 5 min). Although early kinetics of rejoining (within 5 min) was identical for CHO, V79 and repair-deficient TK6 cells, TK6 cells subsequently repaired fewer breaks than did CHO or V79 cells. S phase TK6 cells rejoined SSBs more slowly than cells in the other phases; differences in rejoining rates could not be explained by the presence of a subset of slowly repairing TK6 cells. We conclude that the comet assay detects subtle differences in the tail moment when cells in different phases of the cell cycle are irradiated and allowed to repair damage. The biological importance of these observations remains to be determined.
此前使用多种方法得到的结果表明,辐射诱导的DNA单链断裂(SSB)的诱导或重新连接情况,并不会随细胞周期位置的不同而产生显著差异。然而,通过碱性彗星试验对DNA损伤和细胞周期位置进行同步分析发现,“尾矩”存在细胞周期依赖性差异,而尾矩已被证明是衡量SSB的一个指标。未受辐射的S期细胞显示出显著更高的尾矩,这可能是由于存在活跃复制位点所致。修复一半损伤所需的时间,G1期细胞似乎始终比细胞周期其他阶段的细胞更短(3.7分钟对约5分钟)。尽管CHO、V79和修复缺陷型TK6细胞重新连接的早期动力学(5分钟内)是相同的,但TK6细胞随后修复的断裂比CHO或V79细胞更少。S期TK6细胞重新连接SSB的速度比其他阶段的细胞更慢;重新连接速率的差异无法用存在一小部分修复缓慢的TK6细胞来解释。我们得出结论,彗星试验能够检测出不同细胞周期阶段的细胞受到辐射并进行损伤修复时,尾矩的细微差异。这些观察结果的生物学重要性仍有待确定。