DNA damage induced by the drinking water mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX) in mammalian cells in vitro and in mice.
3-chloro-4-(dichloromethyl)-5-hydroxy-2[5H]-furanone (MX) formed during chlorination of water containing natural organic substances, is a very potent bacterial mutagen. Recently, tumours at multiple sites were reported in rats given MX-containing drinking water. We have investigated the genotoxicity of MX in mammalian cells exposed in vitro and in vivo using alkaline filter elution to detect DNA single-strand breaks and/or alkali-labile sites (SSBs). Concentrations as high as 100 and 300 microM MX were required to induce detectable levels of SSBs in the HL-60 cells. If MX treatment was carried out in the presence of DNA repair inhibitors (AraC plus hydroxyurea), the sensitivity of the assay to detect MX-induced SSBs was increased by a factor of 100. The presence of serum proteins during exposure resulted in a minor reduction of the MX-induced DNA damage in HL-60 cells at the lowest MX concentrations. In primary cultures of testicular cells as well as in resting human peripheral blood mononuclear cells (PBMC), a slightly increased level of SSBs was observed at MX-concentrations above 30 microM, this effect was not further increased by repair inhibitors. In LLC-PK1 renal proximal tubular epithelial cells and in growth stimulated human peripheral PBMC, increased SSBs were detected at MX concentrations as low as low as 3-10 microM and higher using repair inhibitors, and at 10 times higher concentrations without repair inhibitors. No dose dependent DNA damage was detected in the liver, kidney, spleen and colon of male B6C3F1 mice administrated high doses of MX (40 and 80 mg kg-1). Moderately increased and dose dependent SSBs were detected in the liver and kidney in the presence of DNA repair inhibitors during MX treatment, but no such increase was observed in the spleen and colon.
Holme
J A
JA
Department of Environmental Medicine, National Institute of Public Health, P.O. Box 4404 Torshov, N-0403, Oslo, Norway. jorn.holme@folkehelsa.no
Haddeland
U
U
Haug
K
K
Brunborg
G
G
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000187:drug effects; D005663:Furans / Q000633:toxicity*; D018922:HL-60 Cells; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D051381:Rats; D013094:Spermatozoa / Q000187:drug effects; D013552:Swine; D013737:Testis / Q000187:drug effects*; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D014881:Water Supply
D004277:DNA, Single-Stranded; D005663:Furans; D014874:Water Pollutants, Chemical; C054121:3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone