title: Detection of benzylic adducts in DNA and nucleotides from 7-sulfooxymethyl-12-methylbenz[a]anthracene and related compounds by 32P-postlabeling using new TLC systems.
abstract: 7,12-Dimethylbenz[a]anthracene (DMBA) is a highly potent experimental carcinogen, that must be transformed to its ultimate carcinogenic form in vivo. The meso-region theory of aromatic hydrocarbon carcinogenesis predicts that 7-hydroxymethyl sulfate (7-HMBA) ester plays a major role in the metabolic activation, benzylic DNA adduct formation and complete carcinogenicity of HMBA and DMBA. This study was undertaken to detect highly lipophilic benzylic DNA adducts resulting from the reaction between 7-hydroxymethy sulfate ester of HMBA (7-SMBA) and DNA as well as determine their DNA base selectivity. Synthetic 7-SMBA was incubated with DNA (800 microg/ml) and individual deoxynucleoside 3'-monophosphates (600 microg/ml) and benzylic adducts were analyzed by 32P-postlabeling/TLC following their enrichment with butanol extraction. Dilute ammonium hydroxide-based solvents were developed to detect the highly lipophilic aralkyl adducts. The reaction with DNA, dGp and dAp gave rise to multiple adducts; dCp and dTp showed no significant adducts. Chromatographic comparison revealed that the major DNA adduct was derived from dG. The methodology developed was also found applicable for highly lipophilic adducts resulting from sulfate esters of structurally-related metabolites of DMBA.
authors.affiliation: Department of Preventive Medicine and Environmental Health/Toxicology, 354 Health Sciences Research Building, University of Kentucky Medical Center, Lexington, KY 40536, USA.
authors.forename: Manicka V
authors.initials: MV
authors.lastname: Vadhanam
authors.forename: Jamie
authors.initials: J
authors.lastname: Horn
authors.forename: James W
authors.initials: JW
authors.lastname: Flesher
authors.forename: Ramesh C
authors.initials: RC
authors.lastname: Gupta
mesh_terms: D015127:9,10-Dimethyl-1,2-benzanthracene / Q000031:analogs & derivatives* / Q000633:toxicity*; D000818:Animals; D002417:Cattle; D002855:Chromatography, Thin Layer / Q000379:methods*; D018736:DNA Adducts / Q000032:analysis / Q000096:biosynthesis*; D004249:DNA Damage; D003853:Deoxyribonucleosides / Q000032:analysis / Q000378:metabolism; D010761:Phosphorus Radioisotopes
chemical_list: D018736:DNA Adducts; D003853:Deoxyribonucleosides; D010761:Phosphorus Radioisotopes; C018970:7-hydroxymethyl-12-methylbenz(a)anthracene; D015127:9,10-Dimethyl-1,2-benzanthracene