Induction of enzyme activity in cell culture: a rapid screen for detection of planar polychlorinated organic compounds.
Induction of aryl hydrocarbon hydroxylase (AHH) activity in rat hepatoma cell line serves as a simple and rapid method to detect minute (pg) amounts of certain classes of compounds, e.g., dibenzo-p-dioxins, dibenzofurans, and biphenyls. This method may provide a quick screen for such substances in extracts from foods prior to chemical identification. AHH activity is measured by conversion of benzo[a]pyrene (BP) to 3-hydroxy BP in homogenized cell extracts from control and treated cultures and is reported as pmol product formed/mg protein/min. Substances screened by this method include polyhalogenated analogs of dibenzo-p-dioxin (24 compounds), dibenzofuran (11 compounds), biphenyl (7 compounds), and extracts from several food sources. Response of the most reactive compound, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was used to prepare a standard curve, and the AHH activity induced by mole doses of test substance is reported as an ED50 response (the estimated dose needed to produce 50% maximum enzyme induction). The AHH activity induced by food extracts is equated to the standard curve and reported as TCDD equivalents. A potent ED50 response in cell culture appears to correlate well with known toxic responses in other mammalian and avian systems for certain test substances. This correlation suggests that the cell culture enzyme induction method is a useful model for screening food extracts that are suspected to be contaminated with polychlorinated planar substances. A collaborative study would demonstrate the reproducibility of the method.
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis*; D001572:Benzofurans / Q000032:analysis; D001681:Biological Assay; D002478:Cells, Cultured; D004147:Dioxins / Q000032:analysis; D004790:Enzyme Induction / Q000187:drug effects; D005506:Food Contamination / Q000032:analysis; D006843:Hydrocarbons, Chlorinated / Q000032:analysis*; D008114:Liver Neoplasms, Experimental / Q000201:enzymology; D011078:Polychlorinated Biphenyls / Q000032:analysis; D051381:Rats
D001572:Benzofurans; D004147:Dioxins; D006843:Hydrocarbons, Chlorinated; D011078:Polychlorinated Biphenyls; D001189:Aryl Hydrocarbon Hydroxylases