Cytochrome P4501A induction potencies of polycyclic aromatic hydrocarbons in a fish hepatoma cell line:: Demonstration of additive interactions

The relative CYP1A induction potencies, determined as ethoxyresorufin-O-deethylase (EROD) activity, and the cytotoxicities of 19 compounds with one to six benzene rings, mixtures of polycyclic aromatic hydrocarbons (PAHs), and contaminated landfill leachates have been determined in the permanent fish hepatoma cell line PLHC-1. No CYP1A induction was observed with benzene, naphthalene, anthracene, acenaphthene, benzo[g,h,i]perylene, and fluorene and low induction was found with fluoranthene and phenanthrene. All other PAHs with three and more benzene rings led to a concentration-related induction of CYP1A, with rebound decreases at high concentrations resulting in bell-shaped concentration-activity curves. Fish-related induction equivalency factors (IEFs) were estimated for all PAHs on the basis of EC50 values of their EROD activities and are reported here for the first time. The following order of decreasing IEFs was found: dibenz[a,h]anthracene > dibenzo[a,i]pyrene > benzo[k]fluoranthene > 3-methylcholanthrene > benzo[a]pyrene > benzo[e]pyrene > chrysene > 7,1 2-dimethylbenz[a]anthracene > perylene > benz[n]anthracene > pyrene. In contrast to the EROD activity, the immunodetectable protein content determined by ELISA showed a concentration-dependent increase. The interaction of PAHs in mixtures of up to eight individual compounds was additive based on their EROD activities. In landfill leachates, determined induction equivalents (IEQ) were significantly higher than calculated IEQs based on analytical measurements, which indicates additional unknown inducing compounds present in leachates. This study shows that the PLHC-1 cell in vitro system serves as an integrative bioanalytical tool in the ecotoxicological evaluation of aquatic environmental samples contaminated with CYP1A-inducing compounds.