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Potency of isothiocyanates to induce luciferase reporter gene expression via the electrophile-responsive element from murine glutathione S-transferase Ya

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Author: Vermeulen, M. · Boerboom, A.M.M.J.F. · Blankvoort, B.M.G. · Aarts, J.M.M.J.G. · Rietjens, I.M.C.M. · Bladeren, P.J. van · Vaes, W.H.J.
Type:article
Date:2009
Institution: TNO Kwaliteit van Leven
Source:Toxicology in Vitro, 4, 23, 617-621
Identifier: 241580
doi: doi:10.1016/j.tiv.2009.02.005
Keywords: Biology · Analytical research · Antioxidant · Electrophile-responsive element · Isothiocyanate · electrophile · glutathione s transferase Ya · glutathione transferase · isothiocyanic acid derivative · luciferase · phenyl isothiocyanate · sulforaphane · unclassified drug · animal cell · article · cell assay · cell line · chemical structure · concentration response · controlled study · drug potency · enzyme induction · gene expression regulation · gene induction · gene location · mouse · nonhuman · reporter gene · Animals · Cell Line · Cell Survival · Gene Expression Regulation · Genes, Reporter · Glutathione Transferase · Isothiocyanates · Luciferases · Mice · Response Elements · Murinae

Abstract

Isothiocyanates are electrophiles that are able to induce phase II biotransformation enzyme gene expression via an electrophile-responsive element (EpRE) in the gene regulatory region. To study the potency of different isothiocyanates to induce the expression of EpRE-regulated genes, a Hepa-1c1c7 luciferase reporter cell line was exposed to structurally different isothiocyanates. The reporter cell line, EpRE(mGST-Ya)-LUX, contains the EpRE from the regulatory region of the mouse glutathione S-transferase Ya gene. Isothiocyanates containing a methyl-sulfur side chain, e.g. sulforaphane, showed a lower EC50 (0.8-3.2 μM) and a comparable induction factor (17-22.4) compared to the structurally different isothiocyanates containing an alkyl or aromatic side chain, e.g. allyl and phenylethyl isothiocyanate (EC50 3.9-6.5 μM, induction factor 17.5-23). After 24 h of exposure, on average (±SD) 23 ± 5% of the isothiocyanate was found in the cells and 77% in the cell medium. Isothiocyanates prove to be strong inducers of electrophile-responsive element-mediated gene expression at physiological concentrations. The here described luciferase reporter cell line is a suitable assay to measure the potency of compounds to induce EpRE-regulated gene expression. © 2009 Elsevier Ltd. All rights reserved.