Comparison of the formation of benzo[a]pyrene diolepoxide-DNA adducts in vitro by rat and human microsomes: evidence for the involvement of P-450IA1 and P-450IA2

J Biochem Toxicol. 1989 Summer;4(2):79-86. doi: 10.1002/jbt.2570040203.

Abstract

The involvement of cytochrome P-450 isozymes in the activation of benzo[a]pyrene (BP) by human placental and liver microsomes was studied in vitro using monoclonal antibodies (Mab) toward the major 3-methylcholanthrene (MC)-inducible and phenobarbital-inductible rat liver P-450 isozymes (Mab 1-7-1 and Mab 2-66-3, respectively). Microsomes from human placenta and liver and rat liver were incubated with BP and DNA, and BP-diolepoxide-DNA (BPDE-DNA) adducts were measured by synchronous fluorescence spectrophotometry (SFS). The only BP metabolite giving the same fluorescence peak as chemically modified BPDE-DNA was BP-7,8-dihydrodiol. Five (smokers) out of 29 human placentas (smokers and nonsmokers), and five out of nine human livers were able to metabolically activate BP to BPDE-DNA adducts in this system. The Mab 1-7-1 totally inhibited the formation of BPDE-DNA adducts in placental microsomal incubations. Inhibition using rat or human liver microsomes was 50-60% and about 90%, respectively. The Mab 2-66-3 had no effect in any of the microsome types. Adduct formation was inhibited more strongly and at lower concentrations of Mab 1-7-1 compared with the inhibition of AHH activity. This study is a clear indication of the major role of P-450IA1 (P-450c) in human placenta and probably P-450IA2 (P-450d) in human liver in BP activation, while other isozymes also take part in the activation in rat liver. Furthermore, this clearly indicates that AHH activity and BP activation are not necessarily associated.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / analysis*
  • Animals
  • Antibodies, Monoclonal
  • Aryl Hydrocarbon Hydroxylases / metabolism
  • Cytochrome P-450 Enzyme System / biosynthesis
  • Cytochrome P-450 Enzyme System / metabolism*
  • DNA / analysis*
  • DNA Adducts*
  • Dihydroxydihydrobenzopyrenes / analysis*
  • Enzyme Induction / drug effects
  • Female
  • Humans
  • In Vitro Techniques
  • Isoenzymes / biosynthesis
  • Isoenzymes / metabolism*
  • Male
  • Methylcholanthrene / pharmacology
  • Microsomes, Liver / enzymology
  • Microsomes, Liver / metabolism*
  • Phenobarbital / pharmacology
  • Placenta / metabolism
  • Pregnancy
  • Rats
  • Rats, Inbred Strains
  • Species Specificity
  • Spectrometry, Fluorescence

Substances

  • Antibodies, Monoclonal
  • DNA Adducts
  • Dihydroxydihydrobenzopyrenes
  • Isoenzymes
  • benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA
  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
  • Methylcholanthrene
  • DNA
  • Cytochrome P-450 Enzyme System
  • Aryl Hydrocarbon Hydroxylases
  • Phenobarbital