A SIMPLE TECHNIQUE FOR QUANTITATION OF LOW-LEVELS OF DNA DAMAGE IN INDIVIDUAL CELLS
A SIMPLE TECHNIQUE FOR QUANTITATION OF LOW-LEVELS OF DNA DAMAGE IN INDIVIDUAL CELLS
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DOI：
10.1016/0014-4827(88)90265-0
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发表时间：
1988-03-01
影响因子：
3.7
通讯作者：
SCHNEIDER, EL
中科院分区：
文献类型：
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作者：
SINGH, NP;MCCOY, MT;SCHNEIDER, EL
将人淋巴细胞分别置于4℃的x射线照射（26 ~ 200 rad）和H2O2 （9.1 ~ 291 μ m）下，在碱性条件下用单细胞微凝胶电泳技术测定DNA迁移程度。两种药物都诱导了DNA迁移的显著增加，从最低剂量开始评估。在暴露于x射线的细胞中，迁移模式相对均匀，而在H2O2处理的细胞中，迁移模式则不均匀。对暴露于200拉德x射线后的修复动力学进行了分析，并从三个人身上获得淋巴细胞。大部分DNA修复发生在前15分钟内，而所有修复基本上在暴露后120分钟完成。然而，一些细胞在孵育期间没有修复，而另一些细胞显示出DNA迁移模式，表明比初始x射线照射引起的损伤更大。这项技术对于检测单个细胞的损伤和修复似乎是敏感和有用的。
Human lymphocytes were either exposed to X-irradiation (26 to 200 rads) or treated with H2O2 (9.1 to 291 .mu.) at 4.degree.C and the extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Both agents induced a significant increase in DNA migration, beginning at the lowest dose evaluated. Migration patterns are relatively homogeneous among cells exposed to X-rays but heterogeneous among cells treated with H2O2. An analysis of repair kinetics following exposure to 200 rads X-rays was conducted with lymphocytes obtained from three individuals. The bulk of the DNA repair occurred within the first 15 min, while all of the repair was essentially complete by 120 min after exposure. However, some cells demonstrated no repair during this incubation period while other cells demonstrated DNA migration patterns indicative of more damage than that induced by the initial irradiation with X-rays. This technique appears to be sensitive and useful for detecting damage and repair in single cells.