title: Cytochrome P4501A induction and porphyrin accumulation in PLHC-1 fish cells exposed to sediment and oil shale extracts.
abstract: The present study describes the use of a fish hepatoma cell line (PLHC-1) in monitoring the biological effects of sediments collected from recipient waters of the oil shale industry. Sampling sites were located in River Purtse and River Kohtla in northeast Estonia. The effects of pure oil shale on the PLHC-1 cells were also studied. The cells were exposed to n-hexane-extracted samples in 48-well plates for 24 h, and 7-ethoxyresorufin O-deethylase (EROD) activity, total protein, and porphyrin content were measured in the exposed cells. Polycyclic aromatic hydrocarbon (PAH) contents in the samples were measured by high-performance liquid chromatography (HPLC). All the sediment and oil shale samples induced CYP1A activity and led to porphyrin accumulation in the cells. The most potent inducers were the sediments collected near the oil shale processing plants (site Lüganuse in River Purtse and Kohtla in River Kohtla), as well as those at the most downstream site in River Purtse (Purtse). These samples possessed high total PAH contents, ranging from 4,270 to nearly 150,000 microg/kg dry sediment. The presence of other lipophilic organic contaminants in the samples was not determined in this study. Both EROD activity and porphyrin content exhibited biphasic induction curves, and the ED(50)(1) values for EROD activity were lower than the ED(50)s for porphyrin content. 2,3,7, 8-Tetrachlorodibenzo-p-dioxin induction equivalents (TCDD-EQs) calculated from EROD induction potencies correlated well with total PAHs (r(2) = 0.827 and p = 0.003 for log-transformed data) and also with individual PAHs. TCDD-EQs for porphyrin content did not correlate significantly with total PAHs (log-log r(2) = 0.785, p = 0. 116). The biological potency and PAH contamination of the samples showed the same rank order, except at Lüganuse, where sediment extracts induced CYP1A and porphyrins more than could have been expected based on PAH contents. Bioassay-derived induction EQs (normalized to dibenz(a,h)anthracene) were 20- to 3,200-fold greater than EQs calculated from the concentrations of five PAHs, suggesting important contributions from other compounds or nonadditive effects. The PLHC-1 cells proved to be a sensitive bioanalytical tool for sediments contaminated with PAH-type pollutants in the oil shale processing area. We suggest further use of this bioassay in screening and monitoring waters with similar background of pollution as in northeast Estonia.
authors.affiliation: University of Kuopio, Department of Physiology, P.O. Box 1627, FIN-70211 Kuopio, Finland.
authors.forename: S E
authors.initials: SE
authors.lastname: Huuskonen
authors.forename: A
authors.initials: A
authors.lastname: Tuvikene
authors.forename: M
authors.initials: M
authors.lastname: Trapido
authors.forename: K
authors.initials: K
authors.lastname: Fent
authors.forename: M E
authors.initials: ME
authors.lastname: Hahn
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003530:Cyprinidae; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis*; D004784:Environmental Monitoring / Q000379:methods; D004790:Enzyme Induction / Q000187:drug effects; D005069:Evaluation Studies as Topic; D019015:Geologic Sediments; D008114:Liver Neoplasms, Experimental / Q000188:drug therapy* / Q000201:enzymology; D010578:Petroleum / Q000633:toxicity*; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis / Q000633:toxicity*; D011166:Porphyrins / Q000378:metabolism*; D015203:Reproducibility of Results; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D010578:Petroleum; D011084:Polycyclic Aromatic Hydrocarbons; D011166:Porphyrins; D019363:Cytochrome P-450 CYP1A1