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...
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| Format: | Article |
| Language: | English |
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Wiley
2020-01-01
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| Series: | Journal of Nucleic Acids |
| Online Access: | http://dx.doi.org/10.1155/2020/8810105 |
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| author | Donald H. Atha Erdem Coskun Onur Erdem Alessandro Tona Vytas Reipa Bryant C. Nelson |
| author_facet | Donald H. Atha Erdem Coskun Onur Erdem Alessandro Tona Vytas Reipa Bryant C. Nelson |
| author_sort | Donald H. Atha |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-f52cbe19d05d4856ba1c417f4f8f0ec7 |
| institution | Kabale University |
| issn | 2090-0201 2090-021X |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nucleic Acids |
| spelling | doaj-art-f52cbe19d05d4856ba1c417f4f8f0ec72025-02-03T05:44:13ZengWileyJournal of Nucleic Acids2090-02012090-021X2020-01-01202010.1155/2020/88101058810105Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet AssayDonald H. Atha0Erdem Coskun1Onur Erdem2Alessandro Tona3Vytas Reipa4Bryant C. Nelson5National Institute of Standards and Technology, Biosystems and Biomaterials Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USANational Institute of Standards and Technology, Biomolecular Measurement Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USANational Institute of Standards and Technology, Biomolecular Measurement Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USANational Institute of Standards and Technology, Biosystems and Biomaterials Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USANational Institute of Standards and Technology, Biosystems and Biomaterials Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USANational Institute of Standards and Technology, Biosystems and Biomaterials Division, Materials Measurement Laboratory, Gaithersburg, MD 20899, USATo 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.http://dx.doi.org/10.1155/2020/8810105 |
| spellingShingle | Donald H. Atha Erdem Coskun Onur Erdem Alessandro Tona Vytas Reipa Bryant C. Nelson Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay Journal of Nucleic Acids |
| title | Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay |
| title_full | Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay |
| title_fullStr | Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay |
| title_full_unstemmed | Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay |
| title_short | Genotoxic Effects of Etoposide, Bleomycin, and Ethyl Methanesulfonate on Cultured CHO Cells: Analysis by GC-MS/MS and Comet Assay |
| title_sort | genotoxic effects of etoposide bleomycin and ethyl methanesulfonate on cultured cho cells analysis by gc ms ms and comet assay |
| url | http://dx.doi.org/10.1155/2020/8810105 |
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