Methylene chloride-induced DNA damage: an interspecies comparison.
DNA single-strand (ss) breaks were detected in the livers of B6C3F1 mice immediately following exposure to 4000-8000 p.p.m. methylene chloride (MC) for 6 h. This damage was undetectable 2 h after exposure, suggesting an active DNA repair process. Similarly, DNA ss breaks were detected in whole lung homogenates taken from mice exposed to 2000-6000 p.p.m. MC. The DNA of mouse Clara cells incubated in vitro with MC was also damaged at concentrations of 5 mM MC and above. Pre-treatment of mice with the glutathione depletor buthionine sulphoximine (BSO) caused a decrease in the amount of DNA damage detected, suggesting a GST-mediated mechanism. DNA damage was also reduced in Clara cells when incubated in vitro with MC in the presence of BSO. In CHO cells induction of DNA damage was dependent upon exogenous MC metabolism by mouse liver S100 fraction (but not microsomes) in the presence of GSH. DNA ss breaks were not induced by MC in hamster hepatocytes in vitro at concentrations from 5 to 90 mM MC, nor in eight individual samples of normal human hepatocytes exposed to MC at similar concentrations. The ability of MC to induce DNA ss breaks in the four species studied is entirely compatible with the known carcinogenicity of this chemical in animals and offers experimental evidence to suggest that humans would not be susceptible to MC-induced liver cancer. The DNA ss breaks correlate with the metabolism of MC by the GST pathway and provide an explanation for the lack of sensitivity of hamsters and rats to MC-induced liver cancer.
Graves
R J
RJ
Zeneca Central Toxicology Laboratory, Macclesfield, Cheshire, UK.
Coutts
C
C
Green
T
T
D000818:Animals; D019328:Buthionine Sulfoximine; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000502:physiology; D006801:Humans; D008099:Liver / Q000187:drug effects; D008113:Liver Neoplasms / Q000139:chemically induced; D008297:Male; D008647:Mesocricetus; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D008752:Methylene Chloride / Q000633:toxicity*; D051379:Mice; D051381:Rats
D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D008752:Methylene Chloride; D004247:DNA; D005978:Glutathione