Profiles of antioxidant/electrophile response element (ARE/EpRE) nuclear protein binding and c-Ha-ras transactivation in vascular smooth muscle cells treated with oxidative metabolites of benzo[a]pyrene

Kimberly P. Miller, Yun Houng Chen, Victoria L. Hastings, Christopher M. Bral, Kenneth S. Ramos

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Activation of nuclear protein binding to the antioxidant/electrophile response element (ARE/EpRE) by benzo[a]pyrene (BaP) in vascular smooth muscle cells (vSMCs) is associated with transcriptional deregulation of c-Ha-ras. This response may be mediated by oxidative intermediates of BaP generated during the course of cellular metabolism. To test this hypothesis, the profile of ARE/EpRE protein binding and transactivation elicited by BaP was compared with that of 3-hydroxy BaP (3-OH BaP) (0.03 to 3.0 μM), BaP 7,8- dihydrodiol (BaP 7,8-diol) (0.03 to 3.0 μM), BaP 3,6-quinone (BaP 3,6-Q) (0.0003 to 3.0 μM), and H2O2 (25 to 100 μM). Specific protein binding to the consensus c-Ha-ras ARE/EpRE was observed in vSMCs treated with all BaP metabolites at concentrations considerably lower than those required for the parent compound. H2O2, a by-product of BaP 3,6-Q redox cycling, also increased binding to the ARE/EpRE. Treatment of vSMCs with oxidative BaP metabolites or H2O2 transactivated the c-Ha-ras promoter in all instances, but the response was consistently half of the maximal induction elicited by BaP. Similar proteins cross-linked specifically to the consensus c-Ha-ras ARE/EpRE sequence in cells treated with BaP or its oxidative intermediates. The protein binding profile in the c-Ha-ras promoter was similar to that in the NADPH:quinone reductase gene (NQO1) and the glutathione S-transferase Ya gene (GSTYa) promoters, but the relative abundance of individual complexes was promoter-specific. We conclude that oxidative intermediates of BaP mediate activation of nuclear protein binding to ARE/EpRE and contribute to transcriptional de-regulation of c-Ha-ras in vSMCs. (C) 2000 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)1285-1296
Number of pages12
JournalBiochemical Pharmacology
Volume60
Issue number9
DOIs
StatePublished - Nov 1 2000
Externally publishedYes

Keywords

  • Antioxidant/electrophile response element
  • Aromatic hydrocarbons
  • Atherogenesis
  • Gene expression
  • Oxidative stress
  • Vascular injury

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

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