Transactivation of the PPAR-responsive enhancer module in chemopreventive glutathione S-transferase gene by the peroxisome proliferator-activated receptor-gamma and retinoid X receptor heterodimer.
Cancer chemopreventive agents transcriptionally induce glutathione S-transferase (GST), which can protect cells from chemical-induced carcinogenesis. Activation of either NF-E2-related factor-2 (Nrf2) or the CCAAT/enhancer binding protein-beta (C/EBPbeta) contributes to GST induction. Peroxisome proliferator-activated receptor-gamma (PPARgamma) and the retinoic acid X receptor (RXR) play roles in regulating cell differentiation and chemoprevention. This study examined GSTA2 gene induction by the PPARgamma activator and 9-cis-retinoic acid (RA), a RXR ligand, and investigated the molecular basis of PPAR-RXR-mediated GSTA2 induction in the H4IIE hepatocytes. Either 15-deoxy-delta (12, 14)-prostaglandin J(2) (PGJ(2)) or RA induced GSTA2 with Nrf2 and C/EBPbeta activation. When compared with PGJ(2) or RA alone, PGJ(2) + RA enhanced GSTA2 induction, with increases in Nrf2 and C/EBPbeta activation. PGJ(2) + RA increased the luciferase reporter gene activity in the cells transfected with the -1.65-kb flanking region of the GSTA2 gene. Thiazolidinedione PPARgamma agonists, troglitazone, rosiglitazone, and pioglitazone, in combination with RA, potentiated GSTA2 induction, confirming that the activation of the PPARgamma and RXR heterodimer contributed to GSTA2 expression. Deletion of the antioxidant response element- or C/EBP-binding sites or the overexpression of dominant-negative mutant of C/EBP abolished the reporter gene expression. PGJ2 + RA increased the binding of the PPARgamma - RXR heterodimer to the putative PPAR-response elements (PPREs) in the GSTA2 promoter. Specific mutations of these multiple PPRE sites resulted in the complete loss of its responsiveness to PGJ2 + RA, which suggests that these binding sites function as a PPRE-responsive enhancer module (PPREM). Transactivation of PPREM by the PPARgamma - RXR heterodimer was verified by the effective GSTA2 induction in the cells treated with PGJ2 + RA after transfecting them with the plasmids encoding PPARgamma1 and RXRalpha. In conclusion, the PPARgamma - RXR heterodimer promotes GSTA2 induction by activating PPREM in the GSTA2 gene, as well as inducing Nrf2 and C/EBPbeta activation.
Park
Eun Young
EY
National Research Laboratory, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul, Korea.
Cho
Il Je
IJ
Kim
Sang Geon
SG
D000077556:Alitretinoin; D000818:Animals; D001483:Base Sequence; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000096:biosynthesis / Q000235:genetics; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics*; D022781:Hepatocytes; D051379:Mice; D008969:Molecular Sequence Data; D051267:NF-E2-Related Factor 2; D011401:Promoter Regions, Genetic; D015230:Prostaglandin D2 / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists / Q000235:genetics / Q000502:physiology*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000502:physiology*; D020218:Response Elements / Q000502:physiology; D047488:Retinoid X Receptors; D045162:Thiazolidinediones / Q000494:pharmacology; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000819:agonists / Q000235:genetics / Q000502:physiology*; D015533:Transcriptional Activation / Q000187:drug effects / Q000502:physiology; D014212:Tretinoin / Q000494:pharmacology
C097240:15-deoxy-delta(12,14)-prostaglandin J2; D022782:CCAAT-Enhancer-Binding Protein-beta; D004268:DNA-Binding Proteins; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; C495637:Nfe2l2 protein, rat; D018160:Receptors, Cytoplasmic and Nuclear; D018168:Receptors, Retinoic Acid; D047488:Retinoid X Receptors; D045162:Thiazolidinediones; D015534:Trans-Activators; D014157:Transcription Factors; D000077556:Alitretinoin; D014212:Tretinoin; C089946:2,4-thiazolidinedione; D005982:Glutathione Transferase; D015230:Prostaglandin D2