Measurement by 32P-postlabeling of (+/-)anti-benzo[a]pyrene-diolepoxide-N2-deoxyguanosine adduct persistence in unstimulated human peripheral blood lymphocytes

Mutat Res. 1994 Sep;325(1):31-7. doi: 10.1016/0165-7992(94)90024-8.

Abstract

In order to study the relative importance of endogenous and environmental factors for the individual relation between DNA damage and DNA excision repair, a method was developed for measuring quantitatively the persistence of N2-deoxyguanosine adducts formed in non-stimulated isolated human peripheral blood lymphocytes after in vitro incubation with 0.2 microM (+/-)anti-BPDE, applying 32P-postlabeling. Total binding of radiolabeled (+/-)anti-BPDE to DNA and its removal has been studied previously in human peripheral blood lymphocytes, but the method presented here enables the direct investigation of repair of the main (+/-)anti-BPDE-DNA adduct, which is implicated in benzo[a]pyrene-induced mutagenesis. Using this method, it was found that in lymphocytes, obtained from 5 individuals, most (+/-)anti-BPDE-N2-dG adducts are removed within the first 24 h after treatment, while interindividual differences appear to exist in both adduct formation and rate and extent of removal.

MeSH terms

  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / analogs & derivatives*
  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / metabolism*
  • DNA / metabolism*
  • DNA Adducts*
  • DNA Damage*
  • DNA Mutational Analysis / methods
  • DNA Repair / physiology*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / metabolism
  • Humans
  • Lymphocytes / drug effects
  • Lymphocytes / ultrastructure
  • Phosphorus Radioisotopes

Substances

  • DNA Adducts
  • Phosphorus Radioisotopes
  • benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA
  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
  • 7,8-dihydroxy-9,10-epoxide-7,8,9,10-tetrahydrobenzo(a)pyrene-10-deoxyguanosine
  • DNA
  • Deoxyguanosine