Differential responses of PPARalpha, PPARdelta, and PPARgamma reporter cell lines to selective PPAR synthetic ligands.
To characterize the specificity of synthetic compounds for peroxisome proliferator-activated receptors (PPARs), three stable cell lines expressing the ligand binding domain (LBD) of human PPARalpha, PPARdelta, or PPARgamma fused to the yeast GAL4 DNA binding domain (DBD) were developed. These reporter cell lines were generated by a two-step transfection procedure. First, a stable cell line, HG5LN, expressing the reporter gene was developed. These cells were then transfected with the different receptor genes. With the help of the three PPAR reporter cell lines, we assessed the selectivity and activity of PPAR agonists GW7647, WY-14-643, L-165041, GW501516, BRL49653, ciglitazone, and pioglitazone. GW7647, L-165041, and BRL49653 were the most potent and selective agonists for hPPARalpha, hPPARdelta, and hPPARgamma, respectively. Two PPAR antagonists, GW9662 and BADGE, were also tested. GW9662 was a selective PPARgamma antagonist, whereas BADGE was a low-affinity PPAR ligand. Furthermore, GW9662 was a full antagonist on PPARgamma and PPARdelta, whereas it showed partial agonism on PPARalpha. We conclude that our stable models allow specific and sensitive measurement of PPAR ligand activities and are a high-throughput, cell-based screening tool for identifying and characterizing PPAR ligands.
Seimandi
Mathieu
M
INSERM unité 540, Endocrinologie Moléculaire et Cellulaire des Cancers, 60 rue de Navacelles, 34090 Montpellier, France.
Lemaire
Géraldine
G
Pillon
Arnaud
A
Perrin
Agnès
A
Carlavan
Isabelle
I
Voegel
Johannes J
JJ
Vignon
Françoise
F
Nicolas
Jean-Claude
JC
Balaguer
Patrick
P
D000813:Anilides / Q000494:pharmacology; D001559:Benzhydryl Compounds; D002087:Butyrates / Q000494:pharmacology; D004268:DNA-Binding Proteins; D004852:Epoxy Compounds / Q000494:pharmacology; D017930:Genes, Reporter / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D008024:Ligands; D008156:Luciferases / Q000096:biosynthesis; D047493:PPAR alpha / Q000819:agonists / Q000187:drug effects / Q000502:physiology; D047494:PPAR delta / Q000819:agonists / Q000187:drug effects / Q000502:physiology; D047495:PPAR gamma / Q000819:agonists / Q000187:drug effects / Q000502:physiology; D047492:Peroxisome Proliferator-Activated Receptors / Q000819:agonists* / Q000187:drug effects / Q000502:physiology; D010671:Phenylurea Compounds / Q000494:pharmacology; D000077154:Rosiglitazone; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013844:Thiazoles / Q000494:pharmacology; D045162:Thiazolidinediones / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D000098826:Bisphenol A Compounds
D000813:Anilides; D001559:Benzhydryl Compounds; D002087:Butyrates; D004268:DNA-Binding Proteins; D004852:Epoxy Compounds; D008024:Ligands; D008156:Luciferases; D047493:PPAR alpha; D047494:PPAR delta; D047495:PPAR gamma; D047492:Peroxisome Proliferator-Activated Receptors; D010671:Phenylurea Compounds; D000077154:Rosiglitazone; D029701:Saccharomyces cerevisiae Proteins; D013844:Thiazoles; D045162:Thiazolidinediones; D014157:Transcription Factors; C457499:2-chloro-5-nitrobenzanilide; C052130:GAL4 protein, S cerevisiae; C425931:GW 501516; C453899:GW 7647; C019273:bisphenol A diglycidyl ether; D000098826:Bisphenol A Compounds