title: Uranyl acetate induces hprt mutations and uranium-DNA adducts in Chinese hamster ovary EM9 cells.
abstract: Questions about possible adverse health effects from exposures to uranium have arisen as a result of uranium mining, residual mine tailings and use of depleted uranium in the military. The purpose of the current study was to measure the toxicity of depleted uranium as uranyl acetate (UA) in mammalian cells. The activity of UA in the parental CHO AA8 line was compared with that in the XRCC1-deficient CHO EM9 line. Cytotoxicity was measured by clonogenic survival. A dose of 200 microM UA over 24 h produced 3.1-fold greater cell death in the CHO EM9 than the CHO AA8 line, and a dose of 300 microM was 1.7-fold more cytotoxic. Mutagenicity at the hypoxanthine (guanine) phosphoribosyltransferase (hprt) locus was measured by selection with 6-thioguanine. A dose of 200 microM UA produced approximately 5-fold higher averaged induced mutant frequency in the CHO EM9 line relative to the CHO AA8 line. The generation of DNA strand breaks was measured by the alkaline comet assay at 40 min and 24 h exposures. DNA strand breaks were detected in both lines; however a dose response may have been masked by U-DNA adducts or crosslinks. Uranium-DNA adducts were measured by inductively coupled plasma optical emission spectroscopy (ICP-OES) at 24 and 48 h exposures. A maximum adduct level of 8 U atoms/10(3) DNA-P for the 300 microM dose was found in the EM9 line after 48 h. This is the first report of the formation of uranium-DNA adducts and mutations in mammalian cells after direct exposure to a depleted uranium compound. Data suggest that uranium could be chemically genotoxic and mutagenic through the formation of strand breaks and covalent U-DNA adducts. Thus the health risks for uranium exposure could go beyond those for radiation exposure.
authors.affiliation: Department of Chemistry and Biochemistry, Northern Arizona University, PO Box 5698, Flagstaff, AZ 86011-5698, USA. Diane.Stearns@nau.edu
authors.forename: Diane M
authors.initials: DM
authors.lastname: Stearns
authors.forename: Monica
authors.initials: M
authors.lastname: Yazzie
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Bradley
authors.forename: Virginia H
authors.initials: VH
authors.lastname: Coryell
authors.forename: Jake T
authors.initials: JT
authors.lastname: Shelley
authors.forename: Adam
authors.initials: A
authors.lastname: Ashby
authors.forename: Craig S
authors.initials: CS
authors.lastname: Asplund
authors.forename: R Clark
authors.initials: RC
authors.lastname: Lantz
mesh_terms: D000818:Animals; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D006224:Cricetinae; D003412:Cricetulus; D018736:DNA Adducts / Q000187:drug effects* / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects; D043382:DNA-Formamidopyrimidine Glycosylase / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D016296:Mutagenesis / Q000187:drug effects*; D009154:Mutation / Q000187:drug effects* / Q000235:genetics; D009942:Organometallic Compounds / Q000494:pharmacology*; D012259:Ribonuclease, Pancreatic / Q000494:pharmacology; D013866:Thioguanine / Q000494:pharmacology; D014501:Uranium
chemical_list: D018736:DNA Adducts; D009942:Organometallic Compounds; C005460:uranyl acetate; D014501:Uranium; D006861:Hydrogen Peroxide; D007041:Hypoxanthine Phosphoribosyltransferase; D012259:Ribonuclease, Pancreatic; D043382:DNA-Formamidopyrimidine Glycosylase; D013866:Thioguanine