Benzene is a ubiquitous environmental pollutant and a known human leukemogen. Early-life exposure to environmental carcinogens, including benzene, may lead to genomic instability in the fetus, ultimately leading to an increased risk for the development of childhood cancers including leukemia. It is possible that exposure to benzene results in DNA damage that may either be left unrepaired or be repaired erroneously causing genotoxicity.
The first objective of this study was to determine if exposure of fetal hematopoietic cells to p-benzoquinone, a known toxic metabolite of benzene, increased DNA recombination in the pKZ1 model of mutagenesis. A significant increase in recombination was observed following exposure to 25 μM and 50 μM p-benzoquinone for 2, 4, 8, and 24 hours. A significant increase in recombination was also observed following exposure to 25 μM p-benzoquinone for 30 min, 45 min, and 1 hour, but not 15 min as compared to vehicle alone.
Secondly, this study determined if exposure of fetal hematopoietic cells to p-benzoquinone resulted in DNA damage using γ-H2A.X as a marker for DNA double strand breaks and 8-hydroxy-2’-deoxyguanosine as a marker of oxidative DNA damage. A significant increase in γ-H2A.X foci formation was observed following exposure to 25 μM p-benzoquinone for 30 min, 45 min and one hour. Exposure of fetal hematopoietic cells to 25 μM p-benzoquinone did not significantly increase oxidative DNA damage at any of the examined time points.
The third objective of this study was to determine whether or not reactive oxygen species were involved in the observed increase in DNA damage and recombination. Exposure to 25 μM p-benzoquinone for 15 min and 30 min, but not 45 min or one hour, led to an increase in reactive oxygen species production as measured by 5-(and-6)-chloromethyl-2-7-dichlorodihydrofluorescein diacetate fluorescence. Additionally, pretreatment with 400 U/mL PEG-catalase, an antioxidative enzyme, attenuated the increases in both DNA recombination and DNA double strand breaks as compared to treatment with p-benzoquinone alone. These studies indicate that p-benzoquinone is able to induce DNA damage and recombination in fetal hematopoieitic cells and that reactive oxygen species and oxidative stress may be important in the mechanism of toxicity. / Thesis (Master, Pharmacology & Toxicology) -- Queen's University, 2010-07-23 15:44:05.381
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OKQ.1974/5940 |
| Date | 26 July 2010 |
| Creators | MacDonald, Katharine Dawn Dawson |
| Contributors | Queen's University (Kingston, Ont.). Theses (Queen's University (Kingston, Ont.)) |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English, English |
| Detected Language | English |
| Type | Thesis |
| Rights | This publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner. |
| Relation | Canadian theses |
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