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Adipogenic differentiation is not influenced by lentivirus-mediated shRNA targeting the SOCS3 gene in adipose-derived stromal cells

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Abstract

The adipose-derived stromal cells (ADSCs) are capable of adipogenic differentiation by which they can be used as the seed of adipocytes and as the model to study the mechanism of differentiation. Suppressor of cytokine signaling 3 (SOCS3), a negative regulator of leptin signaling, perhaps involves in cell survival and differentiation. However, to date, rare studies are performed on ADSCs differentiation and SOCS3 regulation. In this study, a lentiviral vector expressing a shRNA targeting SOCS3 gene (Lv-shSOCS3) is developed and tested. To observe the effect of Lv-shSOCS3 on ADSCs differentiation, we infected ADSCs with Lv-shSOCS3 or Lv-shSOCS3-NC (non-silenced control lentivirus) and examined adipocyte differentiation using morphology and the expression of key gene for adipogenic differentiation using real-time PCR. The results show that Lv-shSOCS3 can significantly suppress the expression of SOCS3 mRNA and protein in ADSCs. There are no distinguishable differences on adipogenic differentiation and the expression of peroxisome proliferators-activated receptor gamma (PPARγ) mRNA for ADSCs between infection with Lv-shSOCS3 and Lv-shSOCS3-NC. We conclude that SOCS3 knock-down does not influence the growth and adipogenic differentiation feature of ADSCs. Our study provides a new molecular basis for understanding the differentiation mechanism of ADSCs and lays the foundation for further study of the biological functions of SOCS3.

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References

  1. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7:211–228

    Article  CAS  PubMed  Google Scholar 

  2. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295

    Article  CAS  PubMed  Google Scholar 

  3. Rangappa S, Fen C, Lee EH, Bongso A, Sim EK (2003) Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes. Ann Thorac Surg 75:775–779

    Article  PubMed  Google Scholar 

  4. Seo MJ, Suh SY, Bae YC, Jung JS (2005) Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun 328:258–264

    Article  CAS  PubMed  Google Scholar 

  5. Shi H, Tzameli I, Bjorbaek C, Flier JS (2004) Suppressor of cytokine signaling 3 is a physiological regulator of adipocyte insulin signaling. J Biol Chem 279:34733–34740

    Article  CAS  PubMed  Google Scholar 

  6. Yadav A, Kalita A, Dhillon S, Banerjee K (2005) JAK/STAT3 pathway is involved in survival of neurons in response to insulin-like growth factor and negatively regulated by suppressor of cytokine signaling-3. J Biol Chem 280:31830–31840

    Article  CAS  PubMed  Google Scholar 

  7. Wang Z, Zhou YT, Kakuma T, Lee Y, Kalra SP, Kalra PS, Pan W, Unger RH (2000) Leptin resistance of adipocytes in obesity: role of suppressors of cytokine signaling. Biochem Biophys Res Commun 277:20–26

    Article  CAS  PubMed  Google Scholar 

  8. Bantounas I, Phylactou LA, Uney JB (2004) RNA interference and the use of small interfering RNA to study gene function in mammalian systems. J Mol Endocrinol 33:545–557

    Article  CAS  PubMed  Google Scholar 

  9. Aagaard L, Rossi JJ (2007) RNAi therapeutics: principles, prospects and challenges. Adv Drug Deliv Rev 59:75–86

    Article  CAS  PubMed  Google Scholar 

  10. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811

    Article  CAS  PubMed  Google Scholar 

  11. Scherr M, Eder M (2007) Gene silencing by small regulatory RNAs in mammalian cells. Cell Cycle 6:444–449

    CAS  PubMed  Google Scholar 

  12. Pfeifer A, Eigenbrod S, Al-Khadra S, Hofmann A, Mitteregger G, Moser M, Bertsch U, Kretzschmar H (2006) Lentivector-mediated RNAi efficiently suppresses prion protein and prolongs survival of scrapie-infected mice. J Clin Invest 116:3204–3210

    Article  CAS  PubMed  Google Scholar 

  13. Liu L, Zhang Q, Zhang Y, Wang S, Ding Y (2006) Lentivirus-mediated silencing of Tiam1 gene influences multiple functions of a human colorectal cancer cell line. Neoplasia 8:917–924

    Article  CAS  PubMed  Google Scholar 

  14. Katayama K, Wada K, Miyoshi H, Ohashi K, Tachibana M, Furuki R, Mizuguchi H, Hayakawa T, Nakajima A, Kadowaki T, Tsutsumi Y, Nakagawa S, Kamisaki Y, Mayumi T (2004) RNA interfering approach for clarifying the PPARgamma pathway using lentiviral vector expressing short hairpin RNA. FEBS Lett 560:178–182

    Article  CAS  PubMed  Google Scholar 

  15. Gropp M, Reubinoff BE (2007) Lentiviral-RNA-interference system mediating homogenous and monitored level of gene silencing in human embryonic stem cells. Cloning Stem Cells 9:339–345

    Article  CAS  PubMed  Google Scholar 

  16. Zielske SP, Stevenson M (2005) Importin 7 may be dispensable for human immunodeficiency virus type 1 and simian immunodeficiency virus infection of primary macrophages. J Virol 79:11541–11546

    Article  CAS  PubMed  Google Scholar 

  17. Pullmann R Jr, Juhaszova M, Lopez de Silanes I, Kawai T, Mazan-Mamczarz K, Halushka MK, Gorospe M (2005) Enhanced proliferation of cultured human vascular smooth muscle cells linked to increased function of RNA-binding protein HuR. J Biol Chem 280:22819–22826

    Article  CAS  PubMed  Google Scholar 

  18. Sakoda T, Kasahara N, Hamamori Y, Kedes L (1999) A high-titer lentiviral production system mediates efficient transduction of differentiated cells including beating cardiac myocytes. J Mol Cell Cardiol 31:2037–2047

    Article  CAS  PubMed  Google Scholar 

  19. Soneoka Y, Cannon PM, Ramsdale EE, Griffiths JC, Romano G, Kingsman SM, Kingsman AJ (1995) A transient three-plasmid expression system for the production of high titer retroviral vectors. Nucleic Acids Res 23:628–633

    Article  CAS  PubMed  Google Scholar 

  20. Tiscornia G, Singer O, Verma IM (2006) Production and purification of lentiviral vectors. Nat Protoc 1:241–245

    Article  CAS  PubMed  Google Scholar 

  21. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2[-Delta Delta C(T)] method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  22. Bridge AJ, Pebernard S, Ducraux A, Nicoulaz AL, Iggo R (2003) Induction of an interferon response by RNAi vectors in mammalian cells. Nat Genet 34:263–264

    Article  CAS  PubMed  Google Scholar 

  23. Pebernard S, Iggo RD (2004) Determinants of interferon-stimulated gene induction by RNAi vectors. Differentiation 72:103–111

    Article  CAS  PubMed  Google Scholar 

  24. Zhang Z, Zeng B, Jiao G, Li H, Jing Z, Ouyang J, Yuan X, Chai L, Che Y, Zhang Y, Yang R (2009) Suppressor of cytokine signaling 3 promotes bone marrow cells to differentiate into CD8 + T lymphocytes in lung tissue via up-regulating Notch1 expression. Cancer Res 69:1578–1586

    Article  CAS  PubMed  Google Scholar 

  25. Liu YX, Ji L, Yuan HF, Chen L, Xue J, Guan ZX, Nan X, Bai CX, Wang YF, YueW PeiXT (2008) Effect of Silencing SOCS-3 by siRNA on the Erythropoietic Development. Prog Biochem Biophys 35:82–90

    Google Scholar 

  26. Saladin R, Fajas L, Dana S, Halvorsen YD, Auwerx J, Briggs M (1999) Differential regulation of peroxisome proliferator activated receptor gamma1 (PPARgamma1) and PPARgamma2 messenger RNA expression in the early stages of adipogenesis. Cell Growth Differ 10:43–48

    CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by grant from the National Natural Science Foundation Committee of China (No. 30500409).

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Authors and Affiliations

  1. Department of Nutrition and Food Hygiene, School of Public Health, China Medical University, No. 92 Beier Road, Heping District, Shenyang, Liaoning Province, 110001, China

    Li Liu, Jun Yang, Shuang Ma, Fei Yu, Yahao Ren & Li An

  2. Division of Orthopedics, Shengjing Hospital, China Medical University, Heping District, Shenyang, 110004, China

    Hailun Gu

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  1. Li Liu
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  2. Hailun Gu
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Correspondence to Li Liu.

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Liu, L., Gu, H., Yang, J. et al. Adipogenic differentiation is not influenced by lentivirus-mediated shRNA targeting the SOCS3 gene in adipose-derived stromal cells. Mol Biol Rep 37, 2455–2462 (2010). https://doi.org/10.1007/s11033-009-9757-2

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  • DOI: https://doi.org/10.1007/s11033-009-9757-2

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