Improvement of Chemically-activated Luciferase Gene Expression Bioassay for Detection of Dioxin-like Chemicals
Abstract: To improve the chemically-activated luciferase expression (CALUX)bioassay for detection of dioxin-like chemicals (DLCs) based on the toxicity mechanisms of DLCs. Method A recombinant vector was constructed and used to transfect human hepatoma (HepG2). The expression of this vector was 10-100 folds higher than that of pGL2used in previous experiments. The transfected cells showed aromatic hydrocarbon receptor (AhR)-meditated luciferase gene expression. The reliability of luciferase induction in this cell line as a reporter of AhR-mediated toxicity was evaluated, the optimal detection time was examined and a comparison was made by using the commonly used ethoxyresoufin-Odeethylase (EROD) activity induction assay. Result The results suggested that the luciferase activity in recombinant cells was peaked at about 4 h and then decreased to a stable activity by 14 h after TCDD treatment. The detection limit of this cell line was 0.1 lpmol/L, or 10-fold lower than in previous studies, with a linear range from 1 to 100pmol/L, related coefficient of 0.997, and the coefficient of variability (CV) of 15-30%,Conclusion The luciferase induction is 30-fold more sensitive than EROD induction, the detection time is 68 h shorter and the detection procedure is also simpler.
|ZHANG ZHI-REN, XU Shun-qing, ZHOU Yi-kai, XU Yong-jun, LIU ZHI-WEI, CAI XIAo-KUN, TAN XIANG-LIN. Improvement of Chemically-activated Luciferase Gene Expression Bioassay for Detection of Dioxin-like Chemicals[J]. Biomedical and Environmental Sciences, 2002, 15(1): 58-66.