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Forkhead transcription factor Foxo1 is essential for adipocyte differentiation

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Abstract

We analyzed the physiological role of forkhead box class O 1 (Foxo1) in adipocyte differentiation by suppressing Foxo1 mRNA with siRNA specific for Foxo1. Mouse 3T3-L1 preadipocytes infected with an adenovirus expressing Foxo1-siRNA showed a marked decrease in lipid droplet formation when induced to differentiate into adipocytes. Adipocyte differentiation was most severely inhibited by exposing cells to Foxo1-siRNA before induction of differentiation. The incorporation of fluorescent-labeled glucose and fatty acid was significantly inhibited in cells deficient in Foxo1. RT-PCR revealed that downregulation of Foxo1 decreased the expression of the transcription factors, PPAR-γ and C/EBP-α. By comparison, Foxo1-siRNA did not affect the expression of C/EBP-β or C/EBP-δ during the early period of adipocyte differentiation. These results indicate that Foxo1 plays an essential role in adipocyte differentiation, especially at the very early stage of terminal adipocyte differentiation.

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References

  • Abella A.; Dubus P.; Malumbres M.; Rane S. G.; Kiyokawa H.; Sicard A.; Vignon F.; Langin D.; Barbacid M.; Fajas L. Cdk4 promotes adipogenesis through PPARgamma activation. Cell. Metab. 2: 239–249; 2005. doi:10.1016/j.cmet.2005.09.003.

    Article  CAS  PubMed  Google Scholar 

  • Bligh E. G.; Dyer W. J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37: 911–917; 1959.

    CAS  PubMed  Google Scholar 

  • Brunet A.; Bonni A.; Zigmond M. J.; Lin M. Z.; Juo P.; Hu L. S.; Anderson M. J.; Arden K. C.; Blenis J.; Greenberg M. E. Akt promotes cell survival by phosphorylating and inhibiting a forkhead transcription factor. Cell 96: 857–868; 1999. doi:10.1016/S0092-8674(00)80595-4.

    Article  CAS  PubMed  Google Scholar 

  • Brunet A.; Sweeney L. B.; Sturgill J. F.; Chua K. F.; Greer P. L.; Lin Y.; Tran H.; Ross S. E.; Mostoslavsky R.; Cohen H. Y.; Hu L. S.; Cheng H. L.; Jedrychowski M. P.; Gygi S. P.; Sinclair D. A.; Alt F. W.; Greenberg M. E. Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. Science 303: 2011–2015; 2004. doi:10.1126/science.1094637.

    Article  CAS  PubMed  Google Scholar 

  • Cao Z.; Umek R. M.; McKnight S. L. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 5: 1538–1552; 1991. doi:10.1101/gad.5.9.1538.

    Article  CAS  PubMed  Google Scholar 

  • Christian M.; Zhang X.; Schneider-Merck T.; Unterman T. G.; Gellersen B.; White J. O.; Brosens J. J. Cyclic AMP-induced forkhead transcription factor, FKHR, cooperates with CCAAT/enhancer-binding protein beta in differentiating human endometrial stromal cells. J. Biol. Chem. 277: 20825–20832; 2002. doi:10.1074/jbc.M201018200.

    Article  CAS  PubMed  Google Scholar 

  • Classon M.; Kennedy B. K.; Mulloy R.; Harlow E. Opposing roles of pRB and p107 in adipocyte differentiation. Proc. Natl. Acad. Sci. U. S. A. 97: 10826–10831; 2000. doi:10.1073/pnas.190343597.

    Article  CAS  PubMed  Google Scholar 

  • Czech M. P. Insulin’s expanding control of forkheads. Proc. Natl. Acad. Sci. U. S. A. 100: 11198–11200; 2003. doi:10.1073/pnas.2135308100.

    Article  CAS  PubMed  Google Scholar 

  • Fajas L.; Egler V.; Reiter R.; Hansen J.; Kristiansen K.; Debril M. B.; Miard S.; Auwerx J. The retinoblastoma-histone deacetylase 3 complex inhibits PPARgamma and adipocyte differentiation. Dev. Cell. 3: 903–910; 2002a. doi:10.1016/S1534-5807(02)00360-X.

    Article  CAS  PubMed  Google Scholar 

  • Fajas L.; Landsberg R. L.; Huss-Garcia Y.; Sardet C.; Lees J. A.; Auwerx J. E2Fs regulate adipocyte differentiation. Dev. Cell. 3: 39–49; 2002b. doi:10.1016/S1534-5807(02)00190-9.

    Article  CAS  PubMed  Google Scholar 

  • Hansen J. B.; Petersen R. K.; Larsen B. M.; Bartkova J.; Alsner J.; Kristiansen K. Activation of peroxisome proliferator-activated receptor gamma bypasses the function of the retinoblastoma protein in adipocyte differentiation. J. Biol. Chem. 274: 2386–2393; 1999. doi:10.1074/jbc.274.4.2386.

    Article  CAS  PubMed  Google Scholar 

  • Hribal M. L.; Nakae J.; Kitamura T.; Shutter J. R.; Accili D. Regulation of insulin-like growth factor-dependent myoblast differentiation by Foxo forkhead transcription factors. J. Cell. Biol. 162: 535–541; 2003. doi:10.1083/jcb.200212107.

    Article  CAS  PubMed  Google Scholar 

  • Hu E.; Kim J. B.; Sarraf P.; Spiegelman B. M. Inhibition of adipogenesis through MAP kinase-mediated phosphorylation of PPARgamma. Science 274: 2100–2103; 1996. doi:10.1126/science.274.5295.2100.

    Article  CAS  PubMed  Google Scholar 

  • Kamei Y.; Mizukami J.; Miura S.; Suzuki M.; Takahashi N.; Kawada T.; Taniguchi T.; Ezaki O. A forkhead transcription factor FKHR up-regulates lipoprotein lipase expression in skeletal muscle. FEBS Lett. 536: 232–236; 2003. doi:10.1016/S0014-5793(03)00062-0.

    Article  CAS  PubMed  Google Scholar 

  • Kim J. E.; Chen J. Regulation of peroxisome proliferator-activated receptor-gamma activity bymammalian target of rapamycin and amino acids in adipogenesis. Diabetes 53: 2748–2756; 2004. doi:10.2337/diabetes.53.11.2748.

    Article  CAS  PubMed  Google Scholar 

  • Lam E. W.; Francis R. E.; Petkovic M. FOXO transcription factors: key regulators of cell fate. Biochem. Soc. Trans. 34: 722–726; 2006. doi:10.1042/BST0340722.

    Article  CAS  PubMed  Google Scholar 

  • Li P.; Lee H.; Guo S.; Unterman T. G.; Jenster G.; Bai W. AKT-independent protection of prostate cancer cells from apoptosis mediated through complex formation between the androgen receptor and FKHR. Mol. Cell. Biol. 23: 104–118; 2003. doi:10.1128/MCB.23.1.104-118.2003.

    Article  PubMed  Google Scholar 

  • Miki H.; Yamauchi T.; Suzuki R.; Komeda K.; Tsuchida A.; Kubota N.; Terauchi Y.; Kamon J.; Kaburagi Y.; Matsui J.; Akanuma Y.; Nagai R.; Kimura S.; Tobe K.; Kadowaki T. Essential role of insulin receptor substrate 1 (IRS-1) and IRS-2 in adipocyte differentiation. Mol. Cell. Biol. 21: 2521–2532; 2001. doi:10.1128/MCB.21.7.2521-2532.2001.

    Article  CAS  PubMed  Google Scholar 

  • Morrison R. F.; Farmer S. R. Role of PPARgamma in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis. J. Biol. Chem. 274: 17088–17097; 1999. doi:10.1074/jbc.274.24.17088.

    Article  CAS  PubMed  Google Scholar 

  • Naiki T.; Saijou E.; Miyaoka Y.; Sekine K.; Miyajima A. TRB2, a mouse Tribbles ortholog, suppresses adipocyte differentiation by inhibiting AKT and C/EBPbeta. J. Biol. Chem. 282: 24075–24082; 2007. doi:10.1074/jbc.M701409200.

    Article  CAS  PubMed  Google Scholar 

  • Naïmi M.; Gautier N.; Chaussade C.; Valverde A. M.; Accili D.; Van Obberghen E. Dual role of transcription factor FoxO1 in controlling hepatic insulin sensitivity and lipid metabolism. Endocrinology 148: 2424–2434; 2007. doi:10.1210/en.2006-1411.

    Article  PubMed  Google Scholar 

  • Nakae J.; Kitamura T.; Kitamura Y.; Biggs W. H. 3rd; Arden K. C.; Accili D. The forkhead transcription factor Foxo1 regulates adipocyte differentiation. Dev. Cell. 4: 119–129; 2003. doi:10.1016/S1534-5807(02)00401-X.

    Article  CAS  PubMed  Google Scholar 

  • Nakae J.; Kitamura T.; Silver D. L.; Accili D. The forkhead transcription factor Foxo1 (Fkhr) confers insulin sensitivity onto glucose-6-phosphatase expression. J. Clin. Invest. 108: 1359–1367; 2001.

    CAS  PubMed  Google Scholar 

  • Reagan-Shaw S.; Ahmad N. A target for the management of cancer. Toxicol. Appl. Pharmacol. 224: 360–368; 2007. doi:10.1016/j.taap.2006.12.003.

    Article  CAS  PubMed  Google Scholar 

  • Rosen E. D.; Sarraf P.; Troy A. E.; Bradwin G.; Moore K.; Milstone D. S.; Spiegelman B. M.; Mortensen R. M. PPAR gamma is required for the differentiation of adipose tissue in vivo and in vitro. Mol. Cell. 4: 611–617; 1999. doi:10.1016/S1097-2765(00)80211-7.

    Article  CAS  PubMed  Google Scholar 

  • Rosen E. D.; Spiegelman B. M. Adipocytes as regulators of energy balance and glucose homeostasis. Nature 444: 847–853; 2006. doi:10.1038/nature05483.

    Article  CAS  PubMed  Google Scholar 

  • Schmidt M.; Fernandez de Mattos S.; van der Horst A.; Klompmaker R.; Kops G. J.; Lam E. W.; Burgering B. M.; Medema R. H. Cell cycle inhibition by FoxO forkhead transcription factors involves downregulation of cyclin D. Mol. Cell. Biol. 22: 7842–7852; 2002. doi:10.1128/MCB.22.22.7842-7852.2002.

    Article  CAS  PubMed  Google Scholar 

  • Schuster M. B.; Porse B. T. C/EBPalpha: a tumour suppressor in multiple tissues? Biochim. Biophys. Acta 1766: 88–103; 2006.

    CAS  PubMed  Google Scholar 

  • Student A. K.; Hsu R. Y.; Lane M. D. Induction of fatty acid synthetase synthesis in differentiating 3T3-L1 preadipocytes. J. Biol. Chem. 255: 4745–4750; 1980.

    CAS  PubMed  Google Scholar 

  • Tang Q. Q.; Lane M. D. Activation and centromeric localization of CCAAT/enhancer-binding proteins during the mitotic clonal expansion of adipocyte differentiation. Genes Dev. 13: 2231–2241; 1999. doi:10.1101/gad.13.17.2231.

    Article  CAS  PubMed  Google Scholar 

  • Tang Q. Q.; Otto T. C.; Lane M. D. Mitotic clonal expansion: a synchronous process required for adipogenesis. Proc. Natl. Acad. Sci. U. S. A. 100: 44–49; 2003. doi:10.1073/pnas.0137044100.

    Article  CAS  PubMed  Google Scholar 

  • Tran H.; Brunet A.; Grenier J. M.; Datta S. R.; Fornace A. J. Jr.; DiStefano P. S.; Chiang L. W.; Greenberg M. E. DNA repair pathway stimulated by the forkhead transcription factor FOXO3a through the Gadd45 protein. Science 296: 530–534; 2002.

    Article  CAS  PubMed  Google Scholar 

  • Wiper-Bergeron N.; Wu D.; Pope L.; Schild-Poulter C.; Haché R. J. Stimulation of preadipocyte differentiation by steroid through targeting of an HDAC1 complex. EMBO J. 22: 2135–2145; 2003. doi:10.1093/emboj/cdg218.

    Article  CAS  PubMed  Google Scholar 

  • Wu Z.; Bucher N. L.; Farmer S. R. Induction of peroxisome proliferator-activated receptor gamma during the conversion of 3T3 fibroblasts into adipocytes is mediated by C/EBPbeta, C/EBPdelta, and glucocorticoids. Mol. Cell. Biol. 16: 4128–4136; 1996.

    CAS  PubMed  Google Scholar 

  • Wu Z.; Rosen E. D.; Brun R.; Hauser S.; Adelmant G.; Troy A. E.; McKeon C.; Darlington G. J.; Spiegelman B. M. Cross-regulation of C/EBP alpha and PPAR gamma controls the transcriptional pathway of adipogenesis and insulin sensitivity. Mol. Cell. 3: 151–158; 1999. doi:10.1016/S1097-2765(00)80306-8.

    Article  CAS  PubMed  Google Scholar 

  • Zhu Y.; Qi C.; Korenberg J. R.; Chen X. N.; Noya D.; Rao M. S.; Reddy J. K. Structural organization of mouse peroxisome proliferator-activated receptor gamma (mPPAR gamma) gene: alternative promoter use and different splicing yield two mPPAR gamma isoforms. Proc. Natl. Acad. Sci. U. S. A. 92: 7921–7925; 1995. doi:10.1073/pnas.92.17.7921.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We wish to thank Dr. M. Greenberg (Harvard Medical School) for providing expression plasmid of Foxo1, pcDNA3-Foxo1. We are also grateful to Dr. Takiguchi (Chiba University) for providing plasmids pEF-BOS-C/EBP-α, -β, and -δ. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan.

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  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki, Japan

    Keisuke Munekata & Kazuichi Sakamoto

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  1. Keisuke Munekata
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  2. Kazuichi Sakamoto
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Correspondence to Kazuichi Sakamoto.

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Editor: J. Denry Sato

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Munekata, K., Sakamoto, K. Forkhead transcription factor Foxo1 is essential for adipocyte differentiation. In Vitro Cell.Dev.Biol.-Animal 45, 642–651 (2009). https://doi.org/10.1007/s11626-009-9230-5

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