Transitory metabolic disruption and cytotoxicity elicited by benzo[a]pyrene in two cell lines from rainbow trout liver.

Abstract

Two cell lines, RTL-W1 and R1, from rainbow trout liver were used to investigate the effects of benzo[A]pyrene (BaP). BaP induced a catalytic measure of CYP1A, 7-ethoxyresorufin-O-deethylase (EROD) activity, in the rainbow trout liver cell line RTL-W1 but not in R1. Geldanamycin inhibited EROD induction by BaP. Potential BaP metabolites, BaP-7,8-dihydrodiol (BDP) and 6,12-BaP quinone (BQ) also induced EROD activity in RTL-W1. Very low BaP concentrations slightly stimulated cell proliferation in both cell lines. Higher BaP concentrations caused cytotoxicity in RTL-W1 but not in R1. Cytotoxicity was detected in a cell viability assay with 5-carboxyfluorescein diacetate acetoxymethyl ester, and as a decline in cell number. In both cell lines, BaP exposure impaired the reduction of the redox dye, alamar Blue (AB). After BaP removal, AB reduction recovered. Similar results were observed with BQ. As AB monitors metabolic activity, this novel phenomenon was termed transitory metabolic disruption. This decline in AB readings that was caused by BaP was ameliorated in RTL-W1 by alpha-naphthoflavone and geldanamycin, which suggests a role for CYP1A, and in R1 by indomethacin, which suggests involvement of prostaglandin-H-synthase. The significance of the response to BaP that is detected with AB and whether other PAHs cause it will be interesting future questions.