title: Nitroxide TEMPO: a genotoxic and oxidative stress inducer in cultured cells.
abstract: 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO) is a low molecular weight nitroxide and stable free radical. In this study, we investigated the cytotoxicity and genotoxicity of TEMPO in mammalian cells using the mouse lymphoma assay (MLA) and in vitro micronucleus assay. In the absence of metabolic activation (S9), 3mM TEMPO produced significant cytotoxicity and marginal mutagenicity in the MLA; in the presence of S9, treatment of mouse lymphoma cells with 1-2mM TEMPO resulted in dose-dependent decreases of the relative total growth and increases in mutant frequency. Treatment of TK6 human lymphoblastoid cells with 0.9-2.3mM TEMPO increased the frequency of both micronuclei (a marker for clastogenicity) and hypodiploid nuclei (a marker of aneugenicity) in a dose-dependent manner; greater responses were produced in the presence of S9. Within the dose range tested, TEMPO induced reactive oxygen species and decreased glutathione levels in mouse lymphoma cells. In addition, the majority of TEMPO-induced mutants had loss of heterozygosity at the Tk locus, with allele loss of ⩽34Mbp. These results indicate that TEMPO is mutagenic in the MLA and induces micronuclei and hypodiploid nuclei in TK6 cells. Oxidative stress may account for part of the genotoxicity induced by TEMPO in both cell lines.
authors.affiliation: Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Jefferson, AR 72079, United States.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Guo
authors.forename: Roberta A
authors.initials: RA
authors.lastname: Mittelstaedt
authors.forename: Lei
authors.initials: L
authors.lastname: Guo
authors.forename: Joseph G
authors.initials: JG
authors.lastname: Shaddock
authors.forename: Robert H
authors.initials: RH
authors.lastname: Heflich
authors.forename: Anita H
authors.initials: AH
authors.lastname: Bigger
authors.forename: Martha M
authors.initials: MM
authors.lastname: Moore
authors.forename: Nan
authors.initials: N
authors.lastname: Mei
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003497:Cyclic N-Oxides / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism; D051379:Mice; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D003497:Cyclic N-Oxides; D009153:Mutagens; D017382:Reactive Oxygen Species; D005978:Glutathione; C003959:TEMPO