We have previously shown that Hg(2+), Pb(2+), and Cu(2+) significantly induced the expression of Cyp1a1 mRNA, but the catalytic activity was inhibited by the three metals, and the inhibition was accompanied by an increase in the oxidative stress status. In the current study we investigated the role of redox-sensitive transcription factors and the NF-kappaB and AP-1 signaling pathways in the metal-mediated effects on Cyp1a1 gene expression. We show that heavy metals caused the induction of oxidative stress markers, such as reactive oxygen species and heme oxygenase-1, and the depletion of cellular glutathione content, which was associated with NF-kappaB and AP-1 activation. In addition, the NF-kappaB activator PMA significantly abolished the metal-mediated induction of Cyp1a1 mRNA, whereas it further potentiated their inhibitory effects on Cyp1a1 activity. In parallel, the NF-kappaB inhibitor PDTC further potentiated the metal-mediated induction of Cyp1a1 mRNA, whereas it reversed their inhibitory effects on Cyp1a1 activity. Inhibition of AP-1 upstream signaling pathway activators such as JNK by SP600125 suppressed Cyp1a1 mRNA induction by heavy metals, whereas it potentiated their inhibitory effects at the activity level. In contrast, the ERK inhibitor U0126 further potentiated heavy metal-mediated induction of Cyp1a1 mRNA, whereas it reversed their inhibitory effects on the Cyp1a1 activity. The p38 MAPK inhibitor SB203580 suppressed the metal-mediated induction of Cyp1a1 mRNA, but did not alter Cyp1a1 activity. These results clearly demonstrate that activation of the NF-kappaB and AP-1 signaling pathways is directly involved in the modulation of Cyp1a1 by heavy metals.