A biomarker, P450 RGS, for assessing the induction potential of environmental samples

Autor: Robert H. Tukey, Jack W. Anderson, Linda C. Quattrochi, Tien Vu, Steven S. Rossi
Rok vydání: 1995
Předmět:
Zdroj: Environmental Toxicology and Chemistry. 14:1159-1169
ISSN: 1552-8618
0730-7268
Popis: In this study the authors describe the use of a transgenic cell line for the identification of potentially toxic compounds in test solutions and environmental samples. The reporter gene system (RGS), derived from a human liver cancer cell line, has been engineered such that the CYP1A1 gene, when activated by an inducer compound, will produce luciferase instead of P450. Eighteen hours after application of a inducer the reaction is stopped by rinsing, the cells lysed, and the cytoplasm measured for luminescence and protein content (for normalization). Induction by such compounds as dioxin, dioxin-like PCB congeners (coplanars), and polycyclic aromatic hydrocarbons (PAHs) infers these xenobiotics are present at levels that are potentially toxic, carcinogenic, or mutagenic to organisms. Multiple wells with attached cells in 2 ml of media were inoculated with various concentrations to toxicants in organic solvents (including DMSO, toluene, and dichloromethane). Volumes tested successfully thus fare are 2 to 20 {micro}l of solvent. Solvent extracts (using standard extraction methods) of aquatic sediments were directly applied to the assay system. Test results show significant RGS induction from concentrations of inducer compound that, if present in a typical 40-g sediment sample, would be (in ng/g or ppb): 0.05 for dioxin, 10more » to 1,000 for a range of coplanar PCB congeners; about 2,000 for several Aroclor mixtures; and 300 for a mixture of PAHs. A mixture of pesticides at concentrations about four times the PAH levels did not produce an induction response. Results presented here indicate that testing for the presence of potentially toxic chemicals in environmental samples could be performed rapidly, with sensitivity and specificity using this RGS cell line.« less
Databáze: OpenAIRE
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