Donald H Atha, Erdem Coskun, Onur Erdem, Alessandro Tona, Vytas Reipa, Bryant C Nelson
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引用次数: 0
摘要
为了评估分析哺乳动物细胞系基因毒性影响的方法,我们通过单细胞凝胶电泳(彗星试验)和气相色谱-串联质谱(GC-MS/MS)测试了三种常见的基因毒性药物对中国仓鼠卵巢(CHO)细胞的影响。悬浮生长的 CHO 细胞分别与依托泊苷、博莱霉素和甲磺酸乙酯培养,并通过碱性彗星试验和 GC-MS/MS 进行分析。虽然彗星试验能检测到三种药物处理后的 DNA 链断裂,但 GC-MS/MS 只能检测到博莱霉素氧化诱导的 DNA 核碱基损伤。这表明,虽然 GC-MS/MS 在检测基因毒性效应方面存在局限性,但它可用于检测某些导致氧化过程的化学基因毒性物质。彗星试验与气相色谱-质谱/质谱联用,可以成为筛选多种化学基因毒性物质以及监测其他 DNA 损伤因素的更有用的方法。
Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay.
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.