Advertisement

Molecular Biology Reports

, Volume 45, Issue 5, pp 1545–1550 | Cite as

MED31 involved in regulating self-renewal and adipogenesis of human mesenchymal stem cells

  • Erik P. Beadle
  • Joseph A. Straub
  • Bruce A. Bunnell
  • Jamie J. Newman
Short Communication

Abstract

Regulation of gene expression is critical for the maintenance of cell state and homeostasis. Aberrant regulation of genes can lead to unwanted cell proliferation or misdirected differentiation. Here we investigate the role of MED31, a highly conserved subunit of the Mediator complex, to determine the role this subunit plays in the maintenance of human mesenchymal stem cell (hMSC) state. Using siRNA-mediated knockdown of MED31 we demonstrate a decrease in self-renewal based on cell assays and monitoring of gene expression. In addition, in the absence of MED31, hMSCs also displayed a reduction in adipogenesis as evidenced by diminished lipid vesicle formation and expression of specific adipogenic markers. These data present evidence for a significant role for MED31 in maintaining adult stem cell homeostasis, thereby introducing potential novel targets for future investigation and use in better understanding stem cell behavior and adipogenesis.

Keywords

Mediator complex Transcription Adipogenesis Human mesenchymal stem cells 

Notes

Acknowledgements

We would like to thank Dr. Bruce Bunnell and his lab for supplying and supporting training in the culturing and characterization of human mesenchymal stem cells. We thank the Research Core Facility Genomics Core at LSU Health Shreveport, which is supported in part by the Center for Cardiovascular Diseases and Sciences, the Center for Molecular and Tumor Virology, and the Feist-Weiller Cancer Center, for their assistance with microarray hybridization, scanning, and initial data analysis. We would like to acknowledge funding support for this project from the Louisiana Biomedical Research Network, an NIH INBRE grant (8P20GM103424), Louisiana Board of Regents Pilot Funding Program, and Louisiana Tech University College of Applied and Natural Sciences and School of Biological Sciences for support of students and purchasing of supplies. Finally, we would like to thank Matthew Busby and Michael Osmun for their help with primer optimization and preliminary data collection.

Compliance with ethical standards

Conflict of interest

The authors do not have any conflict of interest to disclose.

References

  1. 1.
    Feng J, Mantesso A, De Bari C, Nishiyama A, Sharpe PT (2011) Dual origin of mesenchymal stem cells contributing to organ growth and repair. Proc Natl Acad Sci USA 108:6503–6508CrossRefPubMedGoogle Scholar
  2. 2.
    Le Blanc K, Pittenger MF (2005) Mesenchymal stem cells: progress toward promise. Cytotherapy 7:36–45CrossRefPubMedGoogle Scholar
  3. 3.
    Bartholomew A et al (2002) Mesenchymal stem cells suppress lymphocyte proliferation in vitro and prolong skin graft survival in vivo. Exp Hematol 30:42–48CrossRefPubMedGoogle Scholar
  4. 4.
    Di Nicola M (2002) Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 99:3838–3843CrossRefPubMedGoogle Scholar
  5. 5.
    Blazek E, Mittler G, Meisterernst M (2005) The mediator of RNA polymerase II. Chromosoma 113:399–408CrossRefPubMedGoogle Scholar
  6. 6.
    Kagey MH et al (2010) Mediator and cohesin connect gene expression and chromatin architecture. Nature 467:430–435CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Zhao W et al (2013) SIMPL enhancement of tumor necrosis factor-α dependent p65-MED1 complex formation is required for mammalian hematopoietic stem and progenitor cell function. PLoS ONE 8:e61123CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Crawford SE et al (2002) Defects of the heart, eye, and megakaryocytes in peroxisome proliferator activator receptor-binding protein (PBP) null embryos implicate GATA family of transcription factors. J Biol Chem 277:3585–3592CrossRefPubMedGoogle Scholar
  9. 9.
    Wang X, Yang N, Uno E, Roeder RG, Guo S (2006) A subunit of the mediator complex regulates vertebrate neuronal development. Proc Natl Acad Sci USA 103:17284–17289CrossRefPubMedGoogle Scholar
  10. 10.
    Vogl MR et al (2013) Sox10 cooperates with the mediator subunit 12 during terminal differentiation of myelinating glia. J Neurosci 33:6679–6690CrossRefPubMedGoogle Scholar
  11. 11.
    Schwartz CE et al (2007) The original Lujan syndrome family has a novel missense mutation (p.N1007S) in the MED12 gene. J Med Genet 44:472–477CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Risheg H et al (2007) A recurrent mutation in MED12 leading to R961W causes Opitz-Kaveggia syndrome. Nat Genet 39:451–453CrossRefPubMedGoogle Scholar
  13. 13.
    Vulto-van Silfhout AT et al (2013) Mutations in MED12 cause X-linked Ohdo syndrome. Am J Hum Genet 92:401–406CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Zhou H et al (2012) MED12 mutations link intellectual disability syndromes with dysregulated GLI3-dependent Sonic Hedgehog signaling. Proc Natl Acad Sci USA 109:19763–19768CrossRefPubMedGoogle Scholar
  15. 15.
    Ding N et al (2008) Mediator links epigenetic silencing of neuronal gene expression with X-linked mental retardation. Mol Cell 31:347–359CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Schneider M et al (2015) The nuclear pore-associated TREX-2 complex employs mediator to regulate gene expression. Cell 162:1016–1028CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Tsai K-L et al (2014) Subunit architecture and functional modular rearrangements of the transcriptional mediator complex. Cell 157:1430–1444CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Risley MD, Clowes C, Yu M, Mitchell K, Hentges KE (2010) The Mediator complex protein Med31 is required for embryonic growth and cell proliferation during mammalian development. Dev Biol 342:146–156CrossRefPubMedGoogle Scholar
  19. 19.
    Schiano C, Rienzo M, Casamassimi A, Napoli C (2013) Gene expression profile of the whole Mediator complex in human osteosarcoma and normal osteoblasts. Med Oncol 30:739CrossRefPubMedGoogle Scholar
  20. 20.
    Jiang C, Chen H, Shao L, Wang Q (2014) MicroRNA-1 functions as a potential tumor suppressor in osteosarcoma by targeting Med1 and Med31. Oncol Rep 32:1249–1256CrossRefPubMedGoogle Scholar
  21. 21.
    Huang DW, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4:44–57CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.School of Biological SciencesLouisiana Tech UniversityRustonUSA
  2. 2.Center for Stem Cell Research and Regenerative MedicineTulane University School of MedicineNew OrleansUSA
  3. 3.Departments of PharmacologyTulane University School of MedicineNew OrleansUSA
  4. 4.Division of Regenerative Medicine, Tulane National Primate Research CenterTulane University School of MedicineNew OrleansUSA

Personalised recommendations