Mutagenicity of mercury chloride and mechanisms of cellular defence: the role of metal-binding proteins.
The mechanisms of toxicity and, particularly, the potential carcinogenicity of inorganic mercury are still under debate. Results of mutagenicity and genotoxicity testing with mercury have been inconsistent: mercury induces DNA single-strand breaks at low concentrations in mammalian cells but has not proved mutagenic in several bacterial mutagenicity assays. We investigated the mutagenicity of subtoxic concentrations of inorganic mercury and the role of metal-binding proteins and free radicals in this process. A mutagenicity assay using NIH 3T3 cells, transfected with a vector containing lacZ' as a reporter for mutational events, was applied. In this model, inorganic mercury significantly increased the mutation frequency in the lacZ gene, even at the lowest concentration tested. The mutation frequency was greatest at an Hg(2+) concentration of 0.5 microM. To identify the mechanisms involved, different cellular responses to non-cytotoxic concentrations of HgCl(2) were measured. Hg(2+) increased the intracellular amount of reactive oxygen species. This induction of oxidative stress was observed, although the intracellular glutathione (GSH) and metallothionein (MT) concentrations were increased significantly. Mercury-induced MT expression was even more pronounced after GSH depletion. Correspondingly, radical formation was more evident in the presence of the GSH-depleting agent L-buthioneine-[S:,R:]-sulfoximine. These findings suggest that the observed mutations might be a consequence of oxidative processes, rather than due to a direct interaction of mercury with nuclear DNA. The results also indicate that the auto-induction of MT by Hg(2+) fails to prevent these mutational events.
Schurz
F
F
Department of Analytical and Molecular Pharmacology, TNO Pharma Zeist, Utrecht, The Netherlands.
Sabater-Vilar
M
M
Fink-Gremmels
J
J
D016475:3T3 Cells; D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005609:Free Radicals; D017930:Genes, Reporter; D005978:Glutathione / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007763:Lac Operon; D008627:Mercuric Chloride; D008628:Mercury / Q000378:metabolism; D008668:Metallothionein / Q000378:metabolism; D051379:Mice; D009152:Mutagenicity Tests; D009153:Mutagens; D009154:Mutation; D010100:Oxygen / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism
D000964:Antimetabolites, Antineoplastic; D005609:Free Radicals; D009153:Mutagens; D019328:Buthionine Sulfoximine; D008627:Mercuric Chloride; D008668:Metallothionein; D006861:Hydrogen Peroxide; D008628:Mercury; D005978:Glutathione; D010100:Oxygen