title: Quantification of the mutagenic potency and repair of glycidol-induced DNA lesions.
abstract: Glycidol (Gly) is an electrophilic low-molecular weight epoxide that is classified by IARC as probably carcinogenic to humans. Humans might be exposed to Gly from food, e.g. refined vegetable oils, where Gly has been found as a food process contaminant. It is therefore important to investigate and quantify the genotoxicity of Gly as a primary step towards cancer risk assessment of the human exposure. Here, quantification of the mutagenic potency expressed per dose (AUC: area under the concentration-time curve) of Gly has been performed in Chinese hamster ovary (CHO) cells, using the HPRT assay. The dose of Gly was estimated in the cell exposure medium by trapping Gly with a strong nucleophile, cob(I)alamin, to form stable cobalamin adducts for analysis by LC-MS/MS. Gly was stable in the exposure medium during the time for cell treatment, and thus the dose in vitro is the initial concentration×cell treatment time. Gly induced mutations in the hprt-gene at a rate of 0.08±0.01 mutations/10(5) cells/mMh. Through comparison with the effect of ionizing radiation in the same system a relative mutagenic potency of 9.5rad-eq./mMh was obtained, which could be used for comparison of genotoxicity of chemicals and between test systems and also in procedures for quantitative cancer risk assessment. Gly was shown to induce strand breaks, that were repaired by base excision repair. Furthermore, Gly-induced lesions, present during replication, were found to delay the replication fork elongation. From experiments with repair deficient cells, homologous recombination repair and the ERCC1-XPF complex were indicated to be recruited to support in the repair of the damage related to the stalled replication elongation. The type of DNA damage responsible for the mutagenic effect of Gly could not be concluded from the present study.
authors.affiliation: Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden.
authors.forename: Jenny
authors.initials: J
authors.lastname: Aasa
authors.affiliation: Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, SE-10691 Stockholm, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Vare
authors.affiliation: Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden.
authors.forename: Hitesh V
authors.initials: HV
authors.lastname: Motwani
authors.affiliation: Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, SE-10691 Stockholm, Sweden.
authors.forename: Dag
authors.initials: D
authors.lastname: Jenssen
authors.affiliation: Department of Environmental Science and Analytical Chemistry, Stockholm University, SE-10691 Stockholm, Sweden. Electronic address: margareta.tornqvist@aces.su.se.
authors.forename: Margareta
authors.initials: M
authors.lastname: Törnqvist
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D004852:Epoxy Compounds / Q000633:toxicity*; D005027:Ethylene Oxide / Q000633:toxicity; D009153:Mutagens / Q000633:toxicity; D020005:Propanols / Q000633:toxicity*; D053719:Tandem Mass Spectrometry
chemical_list: D004852:Epoxy Compounds; D009153:Mutagens; D020005:Propanols; C071834:glycidamide; D005027:Ethylene Oxide; C004312:glycidol
keywords: Base excision repair; Glycidol; Mutations; Replication fork elongation; Strand breaks