In vitro and in vivo (32)P-postlabeling analysis of 4-vinyl-1-cyclohexene (butadiene dimer) diepoxide-DNA adducts.
4-Vinyl-1-cyclohexene diepoxide (VCD), and industrial chemical, and its parent compound, 4-vinyl-1-cyclohexene (VCH), are potential health hazards, a they destroy oocytes in follicles in rodents. VCD is also a skin carcinogen at the site of application in both female and male mice and rats and after gavage, induces ovarian tumors in mice and forestomach tumors in rats. A (32)P-postlabeling assay was developed for the detection and measurement of VCD-DNA adducts. VCD, a direct-acting carcinogen, was reacted with DNA in vitro, as well as through mouse skin painting for 3 days with different doses of VCD. (32)P-Labeled adducts were separated by polyethyleneimine (PEI)-cellulose TLC and detected by screen-enhanced autoradiography. Comparable adduct profiles were obtained in vitro and in vivo. At higher doses (36-225 micro mol/mouse), adduct levels in vivo showed a linear dose response, while there was no difference between 14 and 36 micro mol/mouse. The limit of detection was estimated to be 1-3 adducts in 10(8) DNA nucleotides. The results show that VCD exposure gives rise to (presumably pre-mutagenic) DNA adducts in vivo and that (32)P-postlabeling can be applied to biomonitoring of VCD exposure.
Randerath
K
K
Division of Toxicology, Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA.
Mabon
N
N
D000818:Animals; D001345:Autoradiography; D002273:Carcinogens / Q000378:metabolism*; D003510:Cyclohexanes / Q000378:metabolism*; D053138:Cyclohexenes; D018736:DNA Adducts / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D010761:Phosphorus Radioisotopes; D014753:Vinyl Compounds / Q000378:metabolism*
D002273:Carcinogens; D003510:Cyclohexanes; D053138:Cyclohexenes; D018736:DNA Adducts; D010761:Phosphorus Radioisotopes; D014753:Vinyl Compounds; C012606:4-vinyl-1-cyclohexene dioxide