Stable reporter cell lines for peroxisome proliferator-activated receptor gamma (PPARgamma)-mediated modulation of gene expression.

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Gijsbers L, Man HY, Kloet SK, de Haan LH, Keijer J, Rietjens IM, van der Burg B, Aarts JM

Stable reporter cell lines for peroxisome proliferator-activated receptor gamma (PPARgamma)-mediated modulation of gene expression.

Anal Biochem. 2011 Jul 1;414(1):77-83. doi: 10.1016/j.ab.2011.02.032. Epub 2011 Feb 24.

PubMed ID
21354099 [ View in PubMed
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Abstract

Activation of peroxisome proliferator-activated receptor gamma (PPARgamma) by ligands is associated with beneficial health effects, including anti-inflammatory and insulin-sensitizing effects. The aim of the current study was to develop luciferase reporter gene assays to enable fast and low-cost measurement of PPARgamma agonist and antagonist activity. Two reporter gene assays, PPARgamma1 CALUX and PPARgamma2 CALUX, were developed by stable transfection of U2OS cells with an expression vector for PPARgamma1 or PPARgamma2 and a pGL3-3xPPRE-tata-luc or pGL4-3xPPRE-tata-luc reporter construct, respectively. PPARgamma1 CALUX and PPARgamma2 CALUX cells showed similar concentration-dependent luciferase induction upon exposure to the PPARgamma agonists rosiglitazone, troglitazone, pioglitazone, ciglitazone, netoglitazone, and 15-deoxy-Delta(12,14)-prostaglandin J(2). The potency to induce luciferase decreased in the following order: rosiglitazone>troglitazone=pioglitazone>netoglitazone>ciglitazone. A concentration-dependent decrease in the response to 50nM rosiglitazone was observed on the addition of PPARgamma antagonist GW9662 or T0070907 in both PPARgamma1 CALUX and PPARgamma2 CALUX cells. The PPARalpha agonists WY14643 and fenofibrate failed to induce luciferase activity, confirming the specificity of these cell lines for PPARgamma agonists. In conclusion, PPARgamma1 CALUX and PPARgamma2 CALUX cells provide a reliable and useful tool to screen (bio)chemicals for PPARgamma agonist or antagonist activity.