Exposure to benzene, a ubiquitous environmental pollutant, has been linked to leukemogenesis, although the mechanism of benzene initiated carcinogenesis remains unclear. It has been proposed that benzene can be bioactivated to toxic metabolites such as 1,4 benzoquinone (BQ), which can alter signalling pathways and affect chromosomal integrity. BQ has been shown to increase the activity of c-Myb, which is an important transcription factor involved in hematopoiesis, cell proliferation, and cell differentiation. The c-Myb protein also increases topoisomerase IIα (topo IIα) promoter activity specifically in cell lines with hematopoietic origin. Topo II is a critical nuclear enzyme that removes torsional strain by cleaving, untangling and religating double-stranded DNA. Since topo II mediates DNA strand breaks, aberrant topo II activity or increased protein levels may increase the formation of DNA strand breaks, leaving the cell susceptible to mutational events. I hypothesize that BQ increases c-Myb activity, which in turn increases topo IIα promoter activity resulting in increased DNA strand breaks. Using luciferase reporter assays in K-562 cells (human chronic myeloid leukemic cells) I confirmed that BQ exposure (25 and 37 µM) caused an increase in c-Myb activity after 24 hours. Contradictory to previous findings, overexpression of exogenous c-Myb or a polypeptide consisting of c-Myb’s DNA binding domain (DBD), which competitively inhibits the binding of endogenous c-Myb to DNA, did not affect topo IIα promoter activity. However, BQ exposure (37 µM for 24 hours) caused a significant increase in topo IIα promoter activity, which could be blocked by the overexpression of the DBD polypeptide. Western immunoblotting analysis did not show any significant increases in topo IIα protein levels in cells exposed to 37 µM BQ for 24 hours. Overall, this study suggests that BQ exposure increases topo IIα promoter activity through the c-Myb signalling pathway and furthers our understanding of BQ-mediated toxicity. / Thesis (Master, Pharmacology & Toxicology) -- Queen's University, 2008-01-02 14:09:00.011
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OKQ.1974/972 |
| Date | 11 January 2008 |
| Creators | Singh, Roopam |
| 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 |
| Format | 714452 bytes, application/pdf |
| 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 |
Page generated in 0.0026 seconds