Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2003 Dec 15;376(Pt 3):607–613. doi: 10.1042/BJ20030426

Peroxisome-proliferator-activated receptor gamma suppresses Wnt/beta-catenin signalling during adipogenesis.

Marthe Moldes 1, Ying Zuo 1, Ron F Morrison 1, David Silva 1, Bae-Hang Park 1, Jiajian Liu 1, Stephen R Farmer 1
PMCID: PMC1223802  PMID: 12954078

Abstract

The Wnt/beta-catenin signalling pathway appears to operate to maintain the undifferentiated state of preadipocytes by inhibiting adipogenic gene expression. To define the mechanisms regulating suppression of Wnt/beta-catenin signalling, we analysed the beta-catenin expression in response to activation of transcription factors that regulate adipogenesis. The results show an extensive down-regulation of nuclear beta-catenin that occurs during the first few days of differentiation of 3T3-L1 preadipocytes and coincides with the induction of the adipogenic transcription factors, C/EBPbeta (CCAAT-enhancer-binding protein) and PPARgamma (peroxisome-proliferator-activated receptor). To assess the role of each of these factors in this process, we conditionally overexpressed C/EBPbeta in Swiss mouse fibroblasts using the TET-off system. Abundant expression of C/EBPbeta alone had minimal effect on beta-catenin expression, whereas expression of C/EBPbeta, in the presence of dexamethasone, induced PPARgamma expression and caused a measurable decrease in beta-catenin. In addition, exposure of cells expressing both C/EBPbeta and PPARgamma to a potent PPARgamma ligand resulted in an even greater decrease in beta-catenin by mechanisms that involve the proteasome. Our studies also suggest a reciprocal relationship between PPARgamma activity and beta-catenin expression, since ectopic production of Wnt-1 in preadipocytes blocked the induction of PPARgamma gene expression. Moreover, by suppressing beta-catenin expression, ectopic expression of PPARgamma in Wnt-1-expressing preadipocytes rescued the block in adipogenesis after their exposure to the PPARgamma ligand, troglitazone.

Full Text

The Full Text of this article is available as a PDF (217.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altiok S., Xu M., Spiegelman B. M. PPARgamma induces cell cycle withdrawal: inhibition of E2F/DP DNA-binding activity via down-regulation of PP2A. Genes Dev. 1997 Aug 1;11(15):1987–1998. doi: 10.1101/gad.11.15.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Behrens J., Jerchow B. A., Würtele M., Grimm J., Asbrand C., Wirtz R., Kühl M., Wedlich D., Birchmeier W. Functional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta. Science. 1998 Apr 24;280(5363):596–599. doi: 10.1126/science.280.5363.596. [DOI] [PubMed] [Google Scholar]
  3. Bennett Christina N., Ross Sarah E., Longo Kenneth A., Bajnok Laszlo, Hemati Nahid, Johnson Kirk W., Harrison Stephen D., MacDougald Ormond A. Regulation of Wnt signaling during adipogenesis. J Biol Chem. 2002 Jun 7;277(34):30998–31004. doi: 10.1074/jbc.M204527200. [DOI] [PubMed] [Google Scholar]
  4. Cadigan K. M., Nusse R. Wnt signaling: a common theme in animal development. Genes Dev. 1997 Dec 15;11(24):3286–3305. doi: 10.1101/gad.11.24.3286. [DOI] [PubMed] [Google Scholar]
  5. Cao Z., Umek R. M., McKnight S. L. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 1991 Sep;5(9):1538–1552. doi: 10.1101/gad.5.9.1538. [DOI] [PubMed] [Google Scholar]
  6. Desbois-Mouthon C., Cadoret A., Blivet-Van Eggelpoël M. J., Bertrand F., Cherqui G., Perret C., Capeau J. Insulin and IGF-1 stimulate the beta-catenin pathway through two signalling cascades involving GSK-3beta inhibition and Ras activation. Oncogene. 2001 Jan 11;20(2):252–259. doi: 10.1038/sj.onc.1204064. [DOI] [PubMed] [Google Scholar]
  7. Freytag S. O., Geddes T. J. Reciprocal regulation of adipogenesis by Myc and C/EBP alpha. Science. 1992 Apr 17;256(5055):379–382. doi: 10.1126/science.256.5055.379. [DOI] [PubMed] [Google Scholar]
  8. Girnun Geoffrey D., Smith Wendy M., Drori Stavit, Sarraf Pasha, Mueller Elisabetta, Eng Charis, Nambiar Prashant, Rosenberg Daniel W., Bronson Roderick T., Edelmann Winfried. APC-dependent suppression of colon carcinogenesis by PPARgamma. Proc Natl Acad Sci U S A. 2002 Oct 7;99(21):13771–13776. doi: 10.1073/pnas.162480299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Green H., Kehinde O. An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell. 1975 May;5(1):19–27. doi: 10.1016/0092-8674(75)90087-2. [DOI] [PubMed] [Google Scholar]
  10. Hamm J. K., Park B. H., Farmer S. R. A role for C/EBPbeta in regulating peroxisome proliferator-activated receptor gamma activity during adipogenesis in 3T3-L1 preadipocytes. J Biol Chem. 2001 Feb 27;276(21):18464–18471. doi: 10.1074/jbc.M100797200. [DOI] [PubMed] [Google Scholar]
  11. Han Jianrong, Farmer Stephen R., Kirkland James L., Corkey Barbara E., Yoon Richard, Pirtskhalava Tamar, Ido Yasuo, Guo Wen. Octanoate attenuates adipogenesis in 3T3-L1 preadipocytes. J Nutr. 2002 May;132(5):904–910. doi: 10.1093/jn/132.5.904. [DOI] [PubMed] [Google Scholar]
  12. He T. C., Chan T. A., Vogelstein B., Kinzler K. W. PPARdelta is an APC-regulated target of nonsteroidal anti-inflammatory drugs. Cell. 1999 Oct 29;99(3):335–345. doi: 10.1016/s0092-8674(00)81664-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. He T. C., Sparks A. B., Rago C., Hermeking H., Zawel L., da Costa L. T., Morin P. J., Vogelstein B., Kinzler K. W. Identification of c-MYC as a target of the APC pathway. Science. 1998 Sep 4;281(5382):1509–1512. doi: 10.1126/science.281.5382.1509. [DOI] [PubMed] [Google Scholar]
  14. Ikeda S., Kishida S., Yamamoto H., Murai H., Koyama S., Kikuchi A. Axin, a negative regulator of the Wnt signaling pathway, forms a complex with GSK-3beta and beta-catenin and promotes GSK-3beta-dependent phosphorylation of beta-catenin. EMBO J. 1998 Mar 2;17(5):1371–1384. doi: 10.1093/emboj/17.5.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lee Gary, Elwood Fabienne, McNally John, Weiszmann Jennifer, Lindstrom Michelle, Amaral Kate, Nakamura Motonao, Miao Shichang, Cao Ping, Learned R. Marc. T0070907, a selective ligand for peroxisome proliferator-activated receptor gamma, functions as an antagonist of biochemical and cellular activities. J Biol Chem. 2002 Mar 4;277(22):19649–19657. doi: 10.1074/jbc.M200743200. [DOI] [PubMed] [Google Scholar]
  16. Lefebvre A. M., Chen I., Desreumaux P., Najib J., Fruchart J. C., Geboes K., Briggs M., Heyman R., Auwerx J. Activation of the peroxisome proliferator-activated receptor gamma promotes the development of colon tumors in C57BL/6J-APCMin/+ mice. Nat Med. 1998 Sep;4(9):1053–1057. doi: 10.1038/2036. [DOI] [PubMed] [Google Scholar]
  17. Linares-Cruz G., Bruzzoni-Giovanelli H., Alvaro V., Roperch J. P., Tuynder M., Schoevaert D., Nemani M., Prieur S., Lethrosne F., Piouffre L. p21WAF-1 reorganizes the nucleus in tumor suppression. Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):1131–1135. doi: 10.1073/pnas.95.3.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu J., Stevens J., Rote C. A., Yost H. J., Hu Y., Neufeld K. L., White R. L., Matsunami N. Siah-1 mediates a novel beta-catenin degradation pathway linking p53 to the adenomatous polyposis coli protein. Mol Cell. 2001 May;7(5):927–936. doi: 10.1016/s1097-2765(01)00241-6. [DOI] [PubMed] [Google Scholar]
  19. Matsuzawa S. I., Reed J. C. Siah-1, SIP, and Ebi collaborate in a novel pathway for beta-catenin degradation linked to p53 responses. Mol Cell. 2001 May;7(5):915–926. doi: 10.1016/s1097-2765(01)00242-8. [DOI] [PubMed] [Google Scholar]
  20. Morrison R. F., Farmer S. R. Insights into the transcriptional control of adipocyte differentiation. J Cell Biochem. 1999;Suppl 32-33:59–67. doi: 10.1002/(sici)1097-4644(1999)75:32+<59::aid-jcb8>3.3.co;2-t. [DOI] [PubMed] [Google Scholar]
  21. 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. 1999 Jun 11;274(24):17088–17097. doi: 10.1074/jbc.274.24.17088. [DOI] [PubMed] [Google Scholar]
  22. Orford K., Orford C. C., Byers S. W. Exogenous expression of beta-catenin regulates contact inhibition, anchorage-independent growth, anoikis, and radiation-induced cell cycle arrest. J Cell Biol. 1999 Aug 23;146(4):855–868. doi: 10.1083/jcb.146.4.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pear W. S., Nolan G. P., Scott M. L., Baltimore D. Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8392–8396. doi: 10.1073/pnas.90.18.8392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peifer M., Polakis P. Wnt signaling in oncogenesis and embryogenesis--a look outside the nucleus. Science. 2000 Mar 3;287(5458):1606–1609. doi: 10.1126/science.287.5458.1606. [DOI] [PubMed] [Google Scholar]
  25. Polakis P. Wnt signaling and cancer. Genes Dev. 2000 Aug 1;14(15):1837–1851. [PubMed] [Google Scholar]
  26. Roperch J. P., Lethrone F., Prieur S., Piouffre L., Israeli D., Tuynder M., Nemani M., Pasturaud P., Gendron M. C., Dausset J. SIAH-1 promotes apoptosis and tumor suppression through a network involving the regulation of protein folding, unfolding, and trafficking: identification of common effectors with p53 and p21(Waf1). Proc Natl Acad Sci U S A. 1999 Jul 6;96(14):8070–8073. doi: 10.1073/pnas.96.14.8070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosen E. D., Spiegelman B. M. PPARgamma : a nuclear regulator of metabolism, differentiation, and cell growth. J Biol Chem. 2001 Jul 17;276(41):37731–37734. doi: 10.1074/jbc.R100034200. [DOI] [PubMed] [Google Scholar]
  28. Rosen E. D., Walkey C. J., Puigserver P., Spiegelman B. M. Transcriptional regulation of adipogenesis. Genes Dev. 2000 Jun 1;14(11):1293–1307. [PubMed] [Google Scholar]
  29. Ross S. E., Hemati N., Longo K. A., Bennett C. N., Lucas P. C., Erickson R. L., MacDougald O. A. Inhibition of adipogenesis by Wnt signaling. Science. 2000 Aug 11;289(5481):950–953. doi: 10.1126/science.289.5481.950. [DOI] [PubMed] [Google Scholar]
  30. Sadot E., Geiger B., Oren M., Ben-Ze'ev A. Down-regulation of beta-catenin by activated p53. Mol Cell Biol. 2001 Oct;21(20):6768–6781. doi: 10.1128/MCB.21.20.6768-6781.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saez E., Tontonoz P., Nelson M. C., Alvarez J. G., Ming U. T., Baird S. M., Thomazy V. A., Evans R. M. Activators of the nuclear receptor PPARgamma enhance colon polyp formation. Nat Med. 1998 Sep;4(9):1058–1061. doi: 10.1038/2042. [DOI] [PubMed] [Google Scholar]
  32. Shtutman M., Zhurinsky J., Simcha I., Albanese C., D'Amico M., Pestell R., Ben-Ze'ev A. The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Proc Natl Acad Sci U S A. 1999 May 11;96(10):5522–5527. doi: 10.1073/pnas.96.10.5522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tetsu O., McCormick F. Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature. 1999 Apr 1;398(6726):422–426. doi: 10.1038/18884. [DOI] [PubMed] [Google Scholar]
  34. Tetsu O., McCormick F. Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature. 1999 Apr 1;398(6726):422–426. doi: 10.1038/18884. [DOI] [PubMed] [Google Scholar]
  35. Tontonoz P., Hu E., Spiegelman B. M. Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell. 1994 Dec 30;79(7):1147–1156. doi: 10.1016/0092-8674(94)90006-x. [DOI] [PubMed] [Google Scholar]
  36. Willert K., Nusse R. Beta-catenin: a key mediator of Wnt signaling. Curr Opin Genet Dev. 1998 Feb;8(1):95–102. doi: 10.1016/s0959-437x(98)80068-3. [DOI] [PubMed] [Google Scholar]
  37. 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. 1996 Aug;16(8):4128–4136. doi: 10.1128/mcb.16.8.4128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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. 1999 Feb;3(2):151–158. doi: 10.1016/s1097-2765(00)80306-8. [DOI] [PubMed] [Google Scholar]
  39. Wu Z., Xie Y., Bucher N. L., Farmer S. R. Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis. Genes Dev. 1995 Oct 1;9(19):2350–2363. doi: 10.1101/gad.9.19.2350. [DOI] [PubMed] [Google Scholar]
  40. Yeh W. C., Cao Z., Classon M., McKnight S. L. Cascade regulation of terminal adipocyte differentiation by three members of the C/EBP family of leucine zipper proteins. Genes Dev. 1995 Jan 15;9(2):168–181. doi: 10.1101/gad.9.2.168. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES