Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells
BrdU (bromodeoxyuridine) and EdU (ethynyldeoxyuridine) have been largely utilized as the means of monitoring DNA replication and cellular division. Although BrdU induces gene and chromosomal mutations and induces sensitization to photons, EdU‘s effects have not been extensively studied yet. Therefor...
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MDPI AG
2020-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/18/6631 |
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| author | Jeremy S. Haskins Cathy Su Junko Maeda Kade D. Walsh Alexis H. Haskins Allison J. Allum Coral E. Froning Takamitsu A. Kato |
| author_facet | Jeremy S. Haskins Cathy Su Junko Maeda Kade D. Walsh Alexis H. Haskins Allison J. Allum Coral E. Froning Takamitsu A. Kato |
| author_sort | Jeremy S. Haskins |
| collection | DOAJ |
| description | BrdU (bromodeoxyuridine) and EdU (ethynyldeoxyuridine) have been largely utilized as the means of monitoring DNA replication and cellular division. Although BrdU induces gene and chromosomal mutations and induces sensitization to photons, EdU‘s effects have not been extensively studied yet. Therefore, we investigated EdU’s potential cytotoxic and mutagenic effects and its related underlying mechanisms when administered to Chinese hamster ovary (CHO) wild type and DNA repair-deficient cells. EdU treatment displayed a higher cytotoxicity and genotoxicity than BrdU treatment. Cells with defective homologous recombination repair displayed a greater growth delay and severe inhibition of clonogenicity with EdU compared to wild type and other DNA repair-deficient cells. Inductions of sister chromatid exchange and hypoxanthine phosphorybosyl transferase (HPRT) mutation were observed in EdU-incorporated cells as well. Interestingly, on the other hand, EdU did not induce sensitization to photons to the same degree as BrdU. Our results demonstrate that elevated concentrations (similar to manufacturers suggested concentration; >5–10 μM) of EdU treatment were toxic to the cell cultures, particularly in cells with a defect in homologous recombination repair. Therefore, EdU should be administered with additional precautions. |
| first_indexed | 2024-03-10T16:25:22Z |
| format | Article |
| id | doaj.art-4ad0ef60adf946c38dca5009a6f06212 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T16:25:22Z |
| publishDate | 2020-09-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-4ad0ef60adf946c38dca5009a6f062122023-11-20T13:16:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-09-012118663110.3390/ijms21186631Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian CellsJeremy S. Haskins0Cathy Su1Junko Maeda2Kade D. Walsh3Alexis H. Haskins4Allison J. Allum5Coral E. Froning6Takamitsu A. Kato7Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USADepartment of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO 80526, USABrdU (bromodeoxyuridine) and EdU (ethynyldeoxyuridine) have been largely utilized as the means of monitoring DNA replication and cellular division. Although BrdU induces gene and chromosomal mutations and induces sensitization to photons, EdU‘s effects have not been extensively studied yet. Therefore, we investigated EdU’s potential cytotoxic and mutagenic effects and its related underlying mechanisms when administered to Chinese hamster ovary (CHO) wild type and DNA repair-deficient cells. EdU treatment displayed a higher cytotoxicity and genotoxicity than BrdU treatment. Cells with defective homologous recombination repair displayed a greater growth delay and severe inhibition of clonogenicity with EdU compared to wild type and other DNA repair-deficient cells. Inductions of sister chromatid exchange and hypoxanthine phosphorybosyl transferase (HPRT) mutation were observed in EdU-incorporated cells as well. Interestingly, on the other hand, EdU did not induce sensitization to photons to the same degree as BrdU. Our results demonstrate that elevated concentrations (similar to manufacturers suggested concentration; >5–10 μM) of EdU treatment were toxic to the cell cultures, particularly in cells with a defect in homologous recombination repair. Therefore, EdU should be administered with additional precautions.https://www.mdpi.com/1422-0067/21/18/6631EdUBrdUradiosensitizerDNA repair |
| spellingShingle | Jeremy S. Haskins Cathy Su Junko Maeda Kade D. Walsh Alexis H. Haskins Allison J. Allum Coral E. Froning Takamitsu A. Kato Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells International Journal of Molecular Sciences EdU BrdU radiosensitizer DNA repair |
| title | Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells |
| title_full | Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells |
| title_fullStr | Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells |
| title_full_unstemmed | Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells |
| title_short | Evaluating the Genotoxic and Cytotoxic Effects of Thymidine Analogs, 5-Ethynyl-2′-Deoxyuridine and 5-Bromo-2′-Deoxyurdine to Mammalian Cells |
| title_sort | evaluating the genotoxic and cytotoxic effects of thymidine analogs 5 ethynyl 2 deoxyuridine and 5 bromo 2 deoxyurdine to mammalian cells |
| topic | EdU BrdU radiosensitizer DNA repair |
| url | https://www.mdpi.com/1422-0067/21/18/6631 |
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