Atha Donald H, Coskun Erdem, Erdem Onur, Tona Alessandro, Reipa Vytas, Nelson Bryant C
National Institute of Standards and Technology, Biosystems and Biomaterials Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USA.
National Institute of Standards and Technology, Biomolecular Measurement Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USA.
J Nucleic Acids. 2020 Jul 30;2020:8810105. doi: 10.1155/2020/8810105. eCollection 2020.
To evaluate methods for analysis of genotoxic effects on mammalian cell lines, we tested the effect of three common genotoxic agents on Chinese hamster ovary (CHO) cells by single-cell gel electrophoresis (comet assay) and gas chromatography-tandem mass spectrometry (GC-MS/MS). Suspension-grown CHO cells were separately incubated with etoposide, bleomycin, and ethyl methanesulfonate and analyzed by an alkaline comet assay and GC-MS/MS. Although DNA strand breaks were detected by the comet assay after treatment with all three agents, GC-MS/MS could only detect DNA nucleobase lesions oxidatively induced by bleomycin. This demonstrates that although GC-MS/MS has limitations in detection of genotoxic effects, it can be used for selected chemical genotoxins that contribute to oxidizing processes. The comet assay, used in combination with GC-MS/MS, can be a more useful approach to screen a wide range of chemical genotoxins as well as to monitor other DNA-damaging factors.
为了评估分析对哺乳动物细胞系遗传毒性作用的方法，我们通过单细胞凝胶电泳（彗星试验）和气相色谱-串联质谱法（GC-MS/MS）测试了三种常见遗传毒性剂对中国仓鼠卵巢（CHO）细胞的影响。将悬浮培养的CHO细胞分别与依托泊苷、博来霉素和甲磺酸乙酯孵育，并用碱性彗星试验和GC-MS/MS进行分析。尽管在用这三种试剂处理后通过彗星试验检测到了DNA链断裂，但GC-MS/MS只能检测到博来霉素氧化诱导的DNA核碱基损伤。这表明尽管GC-MS/MS在检测遗传毒性作用方面存在局限性，但它可用于某些导致氧化过程的化学遗传毒素。彗星试验与GC-MS/MS结合使用，可能是一种更有用的方法，可用于筛选多种化学遗传毒素以及监测其他DNA损伤因素。
