Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability
Application of DNA damage diagnostic tests is rapidly growing, in particular for ovarian, prostate, and skin cancers; environmental monitoring; chronic and degenerative diseases; and male infertility. Such tests suffer from significant variability among different laboratories due the lack of standar...
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| Format: | Article |
| Language: | English |
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Wiley
2022-01-01
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| Series: | Journal of Nucleic Acids |
| Online Access: | http://dx.doi.org/10.1155/2022/9188636 |
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| author | Donald H. Atha Alessandro Tona Vytas Reipa |
| author_facet | Donald H. Atha Alessandro Tona Vytas Reipa |
| author_sort | Donald H. Atha |
| collection | DOAJ |
| description | Application of DNA damage diagnostic tests is rapidly growing, in particular for ovarian, prostate, and skin cancers; environmental monitoring; chronic and degenerative diseases; and male infertility. Such tests suffer from significant variability among different laboratories due the lack of standardization, experimental validation, and differences in data interpretation. Reference methods and materials for quantitative measurement of UVA-induced DNA damage in mammalian cells are frequently needed. In this study, we examined the use of the single-cell gel electrophoresis (comet) assay to assess the UVA-induced DNA damage in surface-attached Chinese hamster ovary (CHO) cells treated with a photosensitizer as a candidate cellular oxidative damage reference material. We found that the comet images became diffused and the viability of the cells decreased substantially (>20%) as the UVA dose and benzo [a] pyrene (BaP) concentration exceeded 6.3 J/cm2 and 10−6 mol/L BaP. Maintaining the conditions of exposure within this range can improve DNA damage measurement fidelity, particularly if used as a quantitative reference method and to produce materials considered as an in vitro standard for the comet assay. |
| format | Article |
| id | doaj-art-dc73682a252a4574b5c318a54b009fff |
| institution | Kabale University |
| issn | 2090-021X |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Nucleic Acids |
| spelling | doaj-art-dc73682a252a4574b5c318a54b009fff2025-02-03T05:50:35ZengWileyJournal of Nucleic Acids2090-021X2022-01-01202210.1155/2022/9188636Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell ViabilityDonald H. Atha0Alessandro Tona1Vytas Reipa2National Institute of Standards and TechnologyNational Institute of Standards and TechnologyNational Institute of Standards and TechnologyApplication of DNA damage diagnostic tests is rapidly growing, in particular for ovarian, prostate, and skin cancers; environmental monitoring; chronic and degenerative diseases; and male infertility. Such tests suffer from significant variability among different laboratories due the lack of standardization, experimental validation, and differences in data interpretation. Reference methods and materials for quantitative measurement of UVA-induced DNA damage in mammalian cells are frequently needed. In this study, we examined the use of the single-cell gel electrophoresis (comet) assay to assess the UVA-induced DNA damage in surface-attached Chinese hamster ovary (CHO) cells treated with a photosensitizer as a candidate cellular oxidative damage reference material. We found that the comet images became diffused and the viability of the cells decreased substantially (>20%) as the UVA dose and benzo [a] pyrene (BaP) concentration exceeded 6.3 J/cm2 and 10−6 mol/L BaP. Maintaining the conditions of exposure within this range can improve DNA damage measurement fidelity, particularly if used as a quantitative reference method and to produce materials considered as an in vitro standard for the comet assay.http://dx.doi.org/10.1155/2022/9188636 |
| spellingShingle | Donald H. Atha Alessandro Tona Vytas Reipa Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability Journal of Nucleic Acids |
| title | Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability |
| title_full | Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability |
| title_fullStr | Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability |
| title_full_unstemmed | Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability |
| title_short | Development of a Reference Method and Materials for Quantitative Measurement of UV-Induced DNA Damage in Mammalian Cells: Comparison of Comet Assay and Cell Viability |
| title_sort | development of a reference method and materials for quantitative measurement of uv induced dna damage in mammalian cells comparison of comet assay and cell viability |
| url | http://dx.doi.org/10.1155/2022/9188636 |
| work_keys_str_mv | AT donaldhatha developmentofareferencemethodandmaterialsforquantitativemeasurementofuvinduceddnadamageinmammaliancellscomparisonofcometassayandcellviability AT alessandrotona developmentofareferencemethodandmaterialsforquantitativemeasurementofuvinduceddnadamageinmammaliancellscomparisonofcometassayandcellviability AT vytasreipa developmentofareferencemethodandmaterialsforquantitativemeasurementofuvinduceddnadamageinmammaliancellscomparisonofcometassayandcellviability |