Abstract
Reporter genes produce a protein product in transfected cells that can be easily measured in intact or lysed cells and they have been extensively used in numerous basic and applied research applications. Over the past 10 years, reporter gene assays have been widely accepted and used for analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin and related dioxin-like compounds in various types of matrices, such as biological, environmental, food and feed samples, given that high-resolution instrumental analysis techniques are impractical for large-scale screening analysis. The most sensitive cell-based reporter gene bioassay systems developed are the mechanism-based CALUX (Chemically Activated Luciferase Expression) and CAFLUX (Chemically Activated Fluorescent Expression) bioassays, which utilize recombinant cell lines containing stably transfected dioxin (AhR)-responsive firefly luciferase or enhanced green fluorescent protein (EGFP) reporter genes, respectively. While the current CALUX and CAFLUX bioassays are very sensitive, increasing their lower limit of sensitivity, magnitude of response and dynamic range for chemical detection would significantly increase their utility, particularly for those samples that contain low levels of dioxin-like HAHs (i.e., serum). In this study, we report that the addition of modulators of cell signaling pathways or modification of cell culture conditions results in significant improvement in the magnitude and overall responsiveness of the existing CALUX and CAFLUX cell bioassays.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Behnisch PA, Hosoe K, Sakai S. Bioanalytical screening methods for dioxins and dioxin-like compounds a review of bioassay/biomarker technology. Environ Int, 2001, 27: 413–439
Hahn ME. Biomarkers and bioassays for detecting dioxin-like compounds in the marine environment. Sci Total Environ, 2002, 289: 49–69
Denison MS, Zhao B, Baston DS, Clark GC, Murata H, Han DH. Recombinant cell bioassay systems for the detection and relative quantitation of halogenated dioxins and related chemicals. Talanta, 2004, 63: 1123–1133
Whyte JJ, Schmitt CJ, Tillitt DE. The H4IIE cell bioassay as an indicator of dioxin-like chemicals in wildlife and the environment. Crit Rev Toxicol, 2004, 34: 1–83
Harrison RO, Eduliee GH. Immunochemical analysis for dioxinsprogress and prospects. Sci Total Environ, 1999, 239: 1–18
Nagy SR, Sanborn JR, Hammock BD, Denison MS. Development of a green fluorescent protein-based cell bioassay for the rapid and inexpensive detection and characterization of ah receptor agonists. Toxicol Sci, 2002, 65: 200–210
Ziccardi MH, Gardner IA, Denison MS. Development and modification of a recombinant cell bioassay to directly detect halogenated and polycyclic aromatic hydrocarbons in serum. Toxicol Sci, 2000, 54: 183–193
Hoogenboom LA, Hamers AR, Bovee TF. Bioassays for the detection of growth-promoting agents, veterinary drugs and environmental contaminants in food. Analyst, 1999, 124: 79–85
Lai KP, Wong MH, Wong CK. Modulation of AhR-mediated CYP1A1 mRNA and EROD activities by 17beta-estradiol and dexamethasone in TCDD-induced H411E cells. Toxicol Sci, 2004, 78: 41–49
Chen YH, Tukey RH. Protein kinase C modulates regulation of the CYP1A1 gene by the aryl hydrocarbon receptor. J Biol Chem, 1996, 271: 26261–26266
Long WP, Pray-Grant M, Tsai JC, Perdew GH. Protein kinase C activity is required for aryl hydrocarbon receptor pathway-mediated signal transduction. Mol Pharmacol, 1998, 53: 691–700
Brown DJ, Orelien J, Gordon JD, Chu AC, Chu MD, Murata HJ, Kayama F, Denison MS, Clark GC. Mathematical model developed for environmental samples: prediction of GC/MS dioxin TEQ from XDS-CALUX bioassay data. Environ Toxicol Chem, 2007, 41: 4354–4360
Garrison PM, Tullis K, Aarts J, Brouwer A, Giesy JP, Denison MS. Species-specific recombinant cell lines as bioassay systems for the detection of 2,3,7,8-tetrachlorodibenzo-p-dioxin-like chemicals. Fundam Appl Toxicol, 1996, 30: 194–203
Rushing RS, Denison MS. The silencing mediator of retinoic acid and thyroid hormone receptors can interact with the aryl hydrocarbon (Ah) receptor but fails to repress Ah receptor-dependent gene expression. Arch Biochem Biophys, 2002, 403: 189–201
Baggett B, Roy R, Momen S, Morgan S, Tisi L, Morsev D, Gillies RJ. Thermostability of firefly luciferases affects efficiency of detection by in vivo bioluminescence. Mol Imaging, 2004, 3: 324–33
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, B., Baston, D.S., Khan, E. et al. Enhancing the response of CALUX and CAFLUX cell bioassays for quantitative detection of dioxin-like compounds. Sci. China Chem. 53, 1010–1016 (2010). https://doi.org/10.1007/s11426-010-0142-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11426-010-0142-8