title: Uranium induces apoptosis and is genotoxic to normal rat kidney (NRK-52E) proximal cells.
abstract: Uranium (U) is a heavy metal used in the nuclear industry and for military applications. U compounds are toxic. Their toxicity is mediated either by their radioactivity or their chemical properties. Mammalian kidneys and bones are the main organs affected by U toxicity. Although the most characteristic response to U exposure is renal dysfunction, little information is available on the mechanisms of its toxicity at the molecular level. This report studied the genotoxicity of U. Apoptosis induction in normal rat kidney (NRK-52(E)) proximal cells was investigated as a function of exposure time or concentrations (0-800microM). In parallel, DNA damage was evaluated by several methods. In order to distinguish between the intrinsic and the extrinsic pathways of apoptosis, caspases-8, -9, -10 assays were conducted and the mitochondrial membrane potential was measured. Three methods were selected for their complementarities in the detection of genetic lesions. The comet assay was used for the detection of primary lesions of DNA. gamma-H2AX immunostaining was achieved to detect DNA double-strand breaks. The micronucleus assay was used to detect chromosomic breaks or losses. DNA damage and apoptosis were observed in a concentration-dependent manner. This study demonstrated that U is genotoxic from 300microM and induces caspase-dependent apoptosis cell death from 200microM mainly through the intrinsic pathway in NRK-52(E) cells. These results suggest that the DNA damage caused by U is reversible at low concentration (200-400microM) but becomes irreversible and leads to cell death for higher concentrations (500-800microM).
authors.affiliation: Laboratoire Pierre Sue, CEA-CNRS UMR 9956, 91191 Gif sur Yvette, France. celine.thiebault@cea.fr
authors.forename: Céline
authors.initials: C
authors.lastname: Thiébault
authors.forename: Marie
authors.initials: M
authors.lastname: Carrière
authors.forename: Sarah
authors.initials: S
authors.lastname: Milgram
authors.forename: Angélique
authors.initials: A
authors.lastname: Simon
authors.forename: Laure
authors.initials: L
authors.lastname: Avoscan
authors.forename: Barbara
authors.initials: B
authors.lastname: Gouget
mesh_terms: D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D020552:Comet Assay; D004249:DNA Damage; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D015162:Micronucleus Tests; D009153:Mutagens / Q000633:toxicity*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D014501:Uranium / Q000633:toxicity*
chemical_list: D009153:Mutagens; D017382:Reactive Oxygen Species; D014501:Uranium; D053148:Caspase 3; D053453:Caspase 9