The role of the Ah receptor and p38 in benzo[a]pyrene-7,8-dihydrodiol and benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide-induced apoptosis

J Biol Chem. 2003 May 23;278(21):19526-33. doi: 10.1074/jbc.M300780200. Epub 2003 Mar 12.

Abstract

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants in the environment. Benzo[a]pyrene (B[a]P), a prototypical member of this class of chemicals, affects cellular signal transduction pathways and induces apoptosis. In this study, the proximate carcinogen of B[a]P metabolism, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (B[a]P-7,8-dihydrodiol) and the ultimate carcinogen, B[a]P-r-7,t-8-dihydrodiol-t-9,10-epoxide(+/-) (BPDE-2) were found to induce apoptosis in human HepG2 cells. Apoptosis initiated by B[a]P-7,8-dihydrodiol was linked to activation of the Ah receptor and induction of CYP1A1, an event that can lead to the formation of BPDE-2. With both B[a]P-7,8-dihydrodiol and BPDE-2 treatment, changes in anti- and pro-apoptotic events in the Bcl-2 family of proteins correlated with the release of mitochondrial cytochrome c and caspase activation. The onset of apoptosis as monitored by caspase activation was linked to mitogen-activated protein (MAP) kinases. Utilizing mouse hepa1c1c7 cells and the Arnt-deficient BPRc1 cells, activation of MAP kinase p38 by B[a]P-7,8-dihydrodiol was shown to be Ah receptor-dependent, indicating that metabolic activation by CYP1A1 was required. This was in contrast to p38 activation by BPDE-2, an event that was independent of Ah receptor function. Confirmation that MAP kinases play a critical role in BPDE-2-induced apoptosis was shown by inhibiting caspase activation of poly(ADP-ribose)polymerase 1 (PARP-1) by chemical inhibitors of p38 and ERK1/2. Furthermore, mouse embryo p38-/- fibroblasts were shown to be resistant to the actions of BPDE-2-induced apoptosis as determined by annexin V analysis, cytochrome c release, and cleavage of PARP-1. These results confirm that the Ah receptor plays a critical role in B[a]P-7,8-dihydrodiol-induced apoptosis while p38 MAP kinase links the actions of an electrophilic metabolite like BPDE-2 to the regulation of programmed cell death.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / pharmacology*
  • Animals
  • Apoptosis / drug effects*
  • Aryl Hydrocarbon Receptor Nuclear Translocator
  • Carcinogens / pharmacology*
  • Carcinoma, Hepatocellular
  • Caspases / metabolism
  • Cytochrome P-450 CYP1A1 / genetics
  • Cytochrome P-450 CYP1A1 / metabolism
  • Cytochrome c Group / metabolism
  • Cytochrome c Group / pharmacology
  • DNA Damage
  • DNA-Binding Proteins*
  • Dihydroxydihydrobenzopyrenes / pharmacology*
  • Enzyme Activation / drug effects
  • Fibroblasts
  • Humans
  • Liver Neoplasms
  • Luciferases / genetics
  • Mice
  • Mice, Knockout
  • Mitogen-Activated Protein Kinases / deficiency
  • Mitogen-Activated Protein Kinases / physiology*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / analysis
  • Receptors, Aryl Hydrocarbon / physiology*
  • Recombinant Fusion Proteins
  • Transcription Factors / genetics
  • Transfection
  • Tumor Cells, Cultured
  • bcl-X Protein
  • p38 Mitogen-Activated Protein Kinases

Substances

  • ARNT protein, human
  • Arnt protein, mouse
  • BCL2L1 protein, human
  • Bcl2l1 protein, mouse
  • Carcinogens
  • Cytochrome c Group
  • DNA-Binding Proteins
  • Dihydroxydihydrobenzopyrenes
  • Proto-Oncogene Proteins c-bcl-2
  • Receptors, Aryl Hydrocarbon
  • Recombinant Fusion Proteins
  • Transcription Factors
  • bcl-X Protein
  • benzo(a)pyrene 7,8-dihydrodiol
  • Aryl Hydrocarbon Receptor Nuclear Translocator
  • 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
  • Luciferases
  • Cytochrome P-450 CYP1A1
  • Poly(ADP-ribose) Polymerases
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • Caspases