The
benzo(a)pyrene (BaP):
DNA adducts formed in normal human mammary epithelial cell cultures and the
human mammary carcinoma T47D cell line were analyzed by chromatography and
acid hydrolysis of the BaP:deoxyribonucleoside adducts to BaP:
purine adducts and BaP:tetraols. Human mammary epithelial cell cultures and
human mammary carcinoma T47D cells were exposed to [3H]BaP for 24 h, and the levels of binding were 81 and 182 pmol BaP/mg
DNA in normal and T47D cultures, respectively. Analysis of BaP:deoxyribonucleoside adducts resolved by immobilized boronate chromatography and reversephase high-performance liquid chromatography demonstrated the presence of three BaP:deoxyribonucleoside adducts in both cells: M2, MS1, and MS2 in a ratio of 1.6:1:14. Two adducts (MS1 and MS2) bound to the immobilized boronate column indicating the presence of cis-vicinal
hydroxyl groups, a configuration which would result from reaction of 7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7,8,9,10-tetrahydroBaP (
anti-BaPDE) with
DNA. MS2 was identified as (+)-
anti-BaPDE:
deoxyguanosine (dGuo) for it cochromatographed with a [14C]-(+)-
anti-BaPDE:dGuo marker, the BaP:
purine hydrolysis product of MS2 cochromatographed with [14C]-(+)-
anti-BaPDE:
guanine, and the tetraol hydrolysis products cochromatographed with (+/-)-
anti-BaPDE:tetraols. MS1 was identified as (-)-
anti-BaPDE:dGuo for MS1 eluted in the same relative position as a (-)-
anti-BaPDE:dGuo marker, the BaP:
purine hydrolysis product of MS1 cochromatographed with [14C]-(+)-
anti-BaPDE:
guanine, and the tetraol hydrolysis products cochromatographed with (+/-)-
anti-BaPDE:tetraols. Thus, both adducts that bound to the immobilized boronate column were formed from (+/-)-
anti-BaPDE. One major adduct that did not contain cis-vicinal hydroxy groups, M2, was detected in both cell types. M2 was formed from (+/-)-7 beta, 8 alpha-dihydroxy-9 beta, 10 beta-epoxy-7,8,9,10-tetrahydroBaP (syn-BaPDE) as M2 eluted in the same relative position as a syn-BaPDE:dGuo adduct marker and the tetraol hydrolysis products of M2 cochromatographed with tetraols formed from (+/-)-syn-BaPDE. The isolation of the individual BaP:
DNA adducts followed by
acid hydrolysis allowed the identification of the BaP:
DNA adducts formed in human mammary cell cultures and demonstrated the presence of (-)-
anti-BaPDE:dGuo. Thus, this work provides the first evidence, other than cochromatography, that (-)-
anti-BaPDE is formed in cell systems and reacts with
DNA in cells to form (-)-
anti-BaPDE:dGuo.(ABSTRACT TRUNCATED AT 400 WORDS)