TY - JOUR T1 - Activation of <em>c-Ha-ras </em>by Benzo(<em>a</em>)pyrene in Vascular Smooth Muscle Cells Involves Redox Stress and Aryl Hydrocarbon Receptor JF - Molecular Pharmacology JO - Mol Pharmacol SP - 152 LP - 158 DO - 10.1124/mol.58.1.152 VL - 58 IS - 1 AU - J. Kevin Kerzee AU - Kenneth S. Ramos Y1 - 2000/07/01 UR - http://molpharm.aspetjournals.org/content/58/1/152.abstract N2 - Repeated cycles of vascular injury by benzo(a)pyrene (BaP) increase the onset and progression of atherosclerotic lesions in laboratory animals. This atherogenic response is partly mediated by activation of cis-acting antioxidant/electrophile response elements that enhance c-Ha-ras transcription in vascular smooth muscle cells (vSMCs). Activation of antioxidant/electrophile responsive cis-acting elements may depend on metabolism of BaP by cytochrome P450s to intermediates that induce oxidative stress and modulate gene expression. To test this hypothesis, we evaluated mitogen-activated c-Ha-ras expression in vSMCs treated with BaP or its metabolic intermediates alone, and in combination with agents that modulate cellular redox status. BaP (0.3 and 3 μM), BaP-3,6-quinone (0.3 μM), or hydrogen peroxide (50 μM) enhanced serum-activated c-Ha-ras. Ellipticine (0.01 nM), a known inhibitor of cytochrome P450 metabolism and aryl hydrocarbon receptor (AhR) antagonist, inhibitedc-Ha-ras induction by BaP (3 μM). Serum challenge of G0 synchronized cultures of vSMCs withdl-buthionine-(S,R)-sulfoximine (0.1 mM), a depletor of cellular glutathione, increasedc-Ha-ras mRNA levels during the early phase of the mitogenic response. Combined BaP/dl-buthionine-(S,R)-sulfoximine challenge was cytotoxic to the cells and inhibitedc-Ha-ras expression, whereas up-regulation of antioxidant capacity by N-acetylcysteine (0.5 mM) precluded BaP-induced ras expression. BaP increased formation of reactive oxygen species and depleted cellular glutathione, but these changes did not correlate with the kinetics ofc-Ha-ras induction. BaP did not enhancec-Ha-ras expression in vSMCs from AhR knockout mice, although aryl hydrocarbon hydroxylase activity was constitutively expressed in these cells. These results suggest thatc-Ha-ras activation in vSMCs by BaP involves a redox-sensitive mechanism that is coupled to AhR receptor-dependent functions. ER -