Oxidant-induced DNA damage by quartz in alveolar epithelial cells.
Respirable quartz has recently been classified as a human carcinogen. Although, studies with quartz using naked DNA as a target suggest that formation of oxyradicals by particles may play a role in the DNA-damaging properties of quartz, it is not known whether this pathway is important for DNA damage in the target cells for quartz carcinogenesis, i.e. alveolar epithelial cells. Therefore, we determined in vitro DNA damage by DQ12 quartz particles in rat and human and alveolar epithelial cells (RLE, A549) using the single cell gel electrophoresis/comet assay. The radical generation capacity of quartz was analysed by electron spin resonance (ESR) and by immunocytochemical analysis of the hydroxyl radical-specific DNA lesion 8-hydroxydeoxyguanosine (8-OHdG) in the epithelial cells. Quartz particles as well as the positive control hydrogen peroxide, caused a dose-dependent increase in DNA strand breaks in both cell lines. DNA damage by quartz was significantly reduced in the presence of the hydroxyl-radical scavengers mannitol or DMSO. The involvement of hydroxyl radicals was further established by ESR measurements and was also demonstrated by the ability of the quartz to induce formation of 8-OHdG. In conclusion, our data show that quartz elicits DNA damage in rat and human alveolar epithelial cells and indicate that these effects are driven by hydroxyl radical-generating properties of the particles.
Schins
Roel P F
RP
Institut für Umweltmedizinische Forschung, Heinrich-Heine Universität Düsseldorf, Auf'm Hennekamp 50, 40225 Düsseldorf, Germany. roel.schins@uni-duesseldorf.de
Knaapen
Ad M
AM
Cakmak
Gonca D
GD
Shi
Tingming
T
Weishaupt
Christel
C
Borm
Paul J A
PJ
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D002273:Carcinogens; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D020552:Comet Assay; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy; D004847:Epithelial Cells / Q000187:drug effects*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D016877:Oxidants / Q000494:pharmacology*; D011650:Pulmonary Alveoli / Q000187:drug effects*; D011791:Quartz; D051381:Rats
D002273:Carcinogens; D016877:Oxidants; D011791:Quartz; D000080242:8-Hydroxy-2'-Deoxyguanosine; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D003849:Deoxyguanosine