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2010 | 57 | 2 | 209-215
Article title

Evidence for differential effects of glucose and cycloheximide on mRNA levels of peroxisome proliferator-activated receptor- (PPAR-) machinery members: Superinduction of PPAR-γ1 and -γ2 mRNAs

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Quantitative real-time RT-PCR study was conducted to reveal the effects of normal (5 mmol/l) and high (30 mmol/l) glucose without or with oleate (0.3 mmol/l) on mRNA levels of peroxisome proliferator-activated receptor- (PPAR-)α, -γ1, -γ2, and peroxisome proliferator-activated receptor-γ coactivator- (PGC-)1α and -1β in commercial human hepatoma-derived HepG2 cells maintained under low-serum condition. Significant decrease in PPAR-γ1 and PGC-1α mRNA levels to about 50 % was observed during the first 4 h incubation period. During the next 4 h period, both PPAR-γ1 and PGC-1α mRNAs were partly but significantly restored in high glucose batches. In this period, the presence of the transcriptional inhibitor actinomycin D revealed a significant protective effect of excess glucose on mature PPAR-γ1 and PGC-1α mRNAs. Furthermore, PPAR-γ1 and -γ2 mRNAs were differentially superinduced 1.2-2.5 fold in cells upon the administration of the translational inhibitor cycloheximide. When the cells were co-treated with the combination of cycloheximide and actinomycin D, superinduction was completely suppressed, however. Altogether, the experiments revealed, first, an unexpected protective effect of abundant glucose on PPAR-γ1 and PGC-1α mRNAs in HepG2 cells. Second, we demonstrated cycloheximide-induced, transcription-dependent upregulation of mature PPAR-γ1 and -γ2 mRNAs in HepG2 cells associated with preferential expression of the PPAR-γ2 mRNA variant. The results draw attention to as yet unexplored mechanisms involved in the control of PPAR and PGC genes.
Physical description
  • Department of Pathological Physiology, Medical Faculty, Palacky University, Olomouc, Czech Republic
  • Department of Pathological Physiology, Medical Faculty, Palacky University, Olomouc, Czech Republic
  • Blanquart C, Mansouri R, Paumelle R, Fruchart JC, Staels B, Glineur C (2004) The protein kinase C signalling pathway regulates a molecular switch between transactivation and transrepression activity of the peroxisome proliferator-activated receptor alpha (PPARα). Mol Endocrinol 18: 1906-1918.
  • Bogdanova K, Uherkova L, Poczatkova H, Rypka M, Vesely J (2007) mRNA levels of peroxisome proliferator-activated receptors are affected by glucose deprivation and oleate in human hepatoma HepG2 cells. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 151: 237-245.
  • Brown JD, Plutzky J (2007) Peroxisome proliferator-activated receptors as transcriptional nodal points and therapeutic targets. Circulation 115: 518-533.
  • Bustin SA, Benes V, Nolan T, Pfaffl MW (2005) Quantitative real-time RT-PCR - a perspective. J Mol Endocrinol 34: 597-601.
  • Chen Y, Jimenez AR, Medh JD (2006) Identification and regulation of novel PPAR-γ splice variants in human THP-1 macrophages. Biochim Biophys Acta 1759: 32-43.
  • Eberhardt W, Doller A, Akool ES, Pfeilschifter J (2007) Modulation of mRNA stability as a novel therapeutic approach. Pharmacol Ther 114: 56-73.
  • Edwards DR, Mahadevan LC (1992) Protein synthesis inhibitors differentially superinduce c-fos and c-jun by three distinct mechanisms: lack of evidence for labile repressors. EMBO J 11: 2415-2424.
  • Edwardsson U, Ljungberg A, Oscarsson J (2006) Insulin and oleic acid increase PPARγ2 expression in cultured mouse hepatocytes. Biochem Biophys Res Commun 340: 111-117.
  • Fan M, Rhee J, St-Pierre J, Handschin C, Puigserver P, Lin J, Jäeger S, Erdjumend-Bromage H, Tempst P, Spiegelman BM (2004) Suppression of mitochondrial respiration through recruitment of p160 myb binding protein to PGC-1α: modulation by p38 MAPK. Genes Dev 18: 278-289.
  • Finck BN, Kelly DP (2006) PGC-1 coactivators: inducible regulators of energy metabolism in health and disease. J Clin Invest 116: 615-622.
  • Giusti V, Verdumo C, Suter M, Gaillard RC, Burckhardt P, Pralong F (2003) Expression of peroxisome proliferator-activated receptor-γ1 and peroxisome proliferator-activated receptor-γ2 in visceral and subcutaneous adipose tissue of obese women. Diabetes 52: 1673-1676.
  • Handschin C, Rhee J, Lin J, Tarr PT, Spiegelman BM (2003) An autoregulatory loop controls peroxisome proliferator-activated receptor γ coactivator 1α expression in muscle. Proc Natl Acad Sci USA 100: 7111-7116.
  • Handschin C, Lin J, Rhee J, Peyer AK, Chin S, Wu PH, Meyer UA, Spiegelman BM (2005) Nutritional regulation of hepatic heme biosynthesis and porphyria through PGC-1α. Cell 122: 505-515.
  • Handschin C, Spiegelman BM (2006) Peroxisome proliferator-activated receptor γ coactivator 1 coactivators, energy homeostasis, and metabolism. Endocr Rev 27: 728-735.
  • Hershko DD, Robb BW, Wray CJ, Luo GJ, Hasselgren PO (2004) Superinduction of IL-6 by cycloheximide is associated with mRNA stabilization and sustained activation of p38 Map kinase and NF-κB in cultured Cacco-2 cells. J Cell Biochem 91: 951-961.
  • Hu E, Kim JB, Saraf P, Spiegelman BM (1996) Inhibition of adipogenesis through MAP kinase-mediated phosphorylation of PPARγ. Science 274: 2100-2103.
  • Itani OA, Cornish KL, Liu KZ, Thomas CP (2003) Cycloheximide increases glucocorticoid-stimulated α-ENaC mRNA in collecting duct cells by p38 MAPK-dependent pathway. Am J Physiol Renal Physiol 284: F778-F787.
  • Joiakim A, Mathieu PA, Elliott AA, Reiners JJ Jr (2004) Superinduction of CYPA1 in MCF10A cultures by cycloheximide, anisomycin, and puromycin: a process independent of effects on protein translation and unrelated to suppression of aryl hydrocarbon receptor proteolysis by the proteasome. Mol Pharmacol 66: 936-947.
  • Kawaguchi T, Osatomi K, Yamashita H, Kabashima T, Uyeda K (2002) Mechanism for fatty acid 'sparing' effect on glucose-induced transcription regulation of carbohydrate-responsive element-binding protein by AMP-activated protein kinase. J Biol Chem 277: 3829-3835.
  • Kemp TJ, Causton HC, Clerk A (2003) Changes in gene expression induced by H2O2 in cardiac myocytes. Biochem Biophys Res Commun 307: 416-421.
  • Ketzinel M, Kaempfer R (1999) Cell-mediated suppression of human interleukin-2 gene expression at splicing of mRNA. Immunol Lett 68: 161-166.
  • Khan SA, Vanden Heuvel JP (2003) Role of nuclear receptors in the regulation of gene expression by dietary fatty acids. J Nutr Biochem 14: 554-567
  • Liu DC, Zang CB, Liu HY, Possinger K, Fan SG, Elstner E (2004) A novel PPAR α/γ dual agonist inhibits cell growth and induces apoptosis in human glioblastoma T98G cells. Acta Pharmacol Sin 25: 1312-1319.
  • Monsalve M, Wu Z, Adelmant G, Puigserver P, Fan M, Spiegelman BM (2000) Direct coupling of transcription and mRNA processing through the thermogenic coactivator PGC-1. Mol Cell 6: 307-316.
  • Panchapakesan U, Pollock CA, Chen XM (2004) The effect of high glucose and PPAR-γ agonists on PPAR-γ expression and function in HK-2 cells. Am J Physiol Renal Physiol 287: F528-F534.
  • Patel NA, Eichler DC, Chappell DS, Illingworth PA, Chlafant CE, Yamamoto M, Dean NM, Wyatt JR, Mebert K, Watson JE, Cooper DR (2003) The protein kinase C βII exon confers mRNA instability in the presence of high glucose concentrations. J Biol Chem 278: 1149-1157.
  • Poczatkova H, Bogdanova K, Uherkova L, Cervenkova K, Riegrova D, Rypka M, Vesely J (2007) Dehydroepiandrosterone effects on the mRNA levels of peroxisome proliferator-activated receptors and their coactivators in human hepatoma HepG2 cells. Gen Physiol Biophys 26: 268-274.
  • Puigserver P, Spiegelman BM (2003) Peroxisome proliferators-activated receptor-γ coactivator 1α (PGC-1α): transcriptional coactivator and metabolic regulator. Endocr Rev 24: 78-90.
  • Ross J (1995) mRNA Stability in mammalian cells. Microbiol Rev 59: 423-450.
  • Sakata Y, Yoshioka W, Tohyama C, Ohsako S (2007) Internal genomic sequence of human CYP1A1 gene is involved in superinduction of dioxin-induced CYP1A1 transcription by cycloheximide. Biochem Biophys Res Commun 355: 687-692.
  • Semenkovich CF, Loleman T, Goforth R (1993) Physiologic concentration of glucose regulates fatty acid synthase activity in HepG2 cells by mediating fatty acid synthase mRNA stability. J Biol Chem 268: 6961-6970.
  • Semple RK, Chatterjee VK, O'Rahilly S (2006) PPAR gamma and human metabolic disease. J Clin Invest 116: 581-589.
  • Staiger H, Staiger K, Haas C, Weisser M, Machicao F, Haring HU (2005) Fatty acid-induced differential regulation of the genes encoding peroxisome proliferator-activated receptor-gamma coactivator-1α and -1β in human skeletal muscle cells that have been differentiated in vitro. Diabetologia 48: 2115-2118.
  • Tew SR, Hardingham TE (2006) Regulation of SOX9 mRNA in human articular chondrocytes involving p38 MAPK activation and mRNA stabilization. J Biol Chem 281: 39471-39479.
  • Tudor C, Feige JN, Pingali H, Lohray VB, Wahli W, Desvergne B, Engelborhs Y, Gelman L (2007) Association with coregulators is the major determinant governing peroxisome proliferator-activated receptor mobility in living cells. J Biol Chem 282: 4417-4426.
  • Vercauteren K, Pasko RA, Gleyzer N, Marino VM, Scarpulla RC (2006) PGC-1-related coactivator: immediate early expression and characterization of a CREB/NRF-1 binding domain associated with cytochrome c promoter occupancy and respiratory growth. Mol Cell Biol 26: 7409-7419.
  • Yoon JC, Puigserver P, Chen G, Donovan J, Wu Z, Rhee J, Adelmant G, Stafford J, Kahn CR, Granner DK, Newgard CB, Spiegelman BM (2001) Control of hepatic gluconeogenesis through the transcriptional coactivator PGC-1. Nature 413: 131-138.
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