Transcriptional inhibition of TCDD-mediated induction of cytochrome P450 1A1 and alteration of protein expression in a zebrafish hepatic cell line following the administration of TCDD and Cd2

Abstract

We studied the effects of Cd²⁺ on TCDD-mediated induction of the cytochrome P450 1A1 (cyp1a1) gene using a zebrafish liver cell line (ZFL). Our results showed that Cd²⁺ inhibited the TCDD-mediated induction of the cyp1a1 protein, enzyme activity, and mRNA expression level. Cd²⁺ also down-regulated levels of the aryl hydrocarbon receptor (ahr2) and the aryl hydrocarbon receptor nuclear translocator 2b (arnt2b) mRNAs. Compared with TCDD (3nM) treatment alone, co-treatment with Cd²⁺ (0-30μM) and TCDD (3nM) significantly inhibited the activity of the luciferase reporter gene constructs harboring the distal promoter region (P-2626/-2009) of CYP1A1 and the synthetic 3XRE gene promoter. This indicates that Cd²⁺ decreased the level of TCDD-induced cyp1a1 through transcriptional inhibition. Proteomic analysis was also used to evaluate the effect of Cd²⁺ on TCDD-altered protein expression in ZFL cells. The identified proteins are mainly enzymes of the glycolysis pathway and proteasomes, and have anti-oxidative and anti-stress effects.

Keywords: Aryl hydrocarbon receptor; Gene transcription; Proteomics.