N-COR MODULATES OSTEOGENIC DIFFERENTIATION OF RAT MESENCHYMAL STEM CELLS THROUGH THE PI3K/AKT-CELL SIGNALING PATHWAY

Authors

  • Yi Qin 1. Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120 2. Department of Orthopedics, Zhuhai People’s Hospital, Zhuhai 519000
  • Guoping Cao Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405
  • Jichao Ye Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120
  • Peng Wang Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120
  • Liangbin Gao Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120
  • Suwei Wang Department of Orthopedics, Zhuhai People’s Hospital, Zhuhai 519000
  • Huiyong Shen Department of Orthopedics, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120

Abstract

The nuclear receptor corepressor (N-CoR) is involved in the regulation of diverse transcription factors. We previously found that N-CoR could regulate adipogenic differentiation of rat mesenchymal stem cells (MSCs), but whether it modulated osteogenic differentiation of this type of cells was uncertain. Therefore, in the present study, we investigated the effect and mechanism of N-CoR on osteogenic differentiation of rat MSCs. The results showed that MSCs cultured in osteogenic medium successfully differentiated into osteogenic cells. Overexpression of N-CoR decreased cell proliferation, alkaline phosphatase (ALP) activity, calcium accumulation, mRNA expression of genes including bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), Osterix and Runx2, and protein expression of phosphor-Akt (pAkt). Conversely, knocking down cellular N-CoR by small interfering RNA (siRNA) promoted pAkt activity and cell differentiation. Overexpression or knockdown of N-CoR had no significant influences on the protein expression of pyruvate dehydrogenase kinase isozyme 1 (PDK1), phosphatidylinositol 3-kinase (PI3K) and total Akt, indicating that N-CoR regulated the changes in the PI3K/Akt signaling pathway by targeting pAkt. To further prove the function of the PI3K/Akt signaling in N-CoR-regulated osteogenic differentiation, we used the PI3K inhibitor (LY294002) to block the activation of the PI3K/Akt pathway and found that overexpression of N-CoR showed no effects on ALP activity, calcium level and mRNA expression of BSP, osteocalcin OCN, OPN, Osterix and Runx2 in rat MSCs following the inhibition of the PI3K/Akt pathway. These findings suggest that N-CoR regulates osteogenic differentiation of rat MSCs through suppression of the PI3K/Akt signaling pathway.

 

Key words: N-CoR; MSCs; osteogenic differentiation; PI3K; Akt

 

Received: September 1, 2015; Revised: October 16, 2015; Accepted: October 21, 2015; Published online: April 27, 2016

Downloads

Download data is not yet available.

References

Ding DC, Shyu WC, Lin SZ. Mesenchymal stem cells. Cell Transplant. 2011;20:5-14.

Ma L, Feng XY, Cui BL, Law F, Jiang XW, Yang LY, Xie QD, Huang TH. Human umbilical cord Wharton’s Jelly-derived mesenchymal stem cells differentiation into nerve-like cells. Chin Med J (Engl). 2005;118:1987-93.

Caplan AI. Mesenchymal stem cells. J Ortho Res. 1991;9:641-50.

Aubin J, Triffitt JT. Mesenchymal stem cell and osteoblast Differentiation. In: Bilezikian J, Raisz LG, Rodan GA, editors. Principles of bone biology. San Diego, CA, USA; 2002. p. 59-82.

Hermanson O, Jepsen K, Rosenfeld MG. N-CoR controls differentiation of neural stem cells into astrocytes. Nature. 2002;419:934-9.

Sardi SP, Murtie J, Koirala S, Patten BA, Corfas G. Presenilin-dependent ErbB4 nuclear signaling regulates the timing of astrogenesis in the developing brain. Cell. 2006;127:185-97.

Horlein AJ, Naar AM, Heinzel T, Torchia J, Gloss B, Kurokawa R. Ligand-independent repression by the thyroid hormone receptor mediated by a nuclear receptor co-repressor. Nature. 1995;377:397-404.

Furuya F, Guigon CJ, Zhao L, Lu C, Hanover JA, Cheng SY. Nuclear receptor corepressor is a novel regulator of phosphatidylinositol 3-kinase signaling. Mol Cell Biol. 2007;27(17): 6116-26.

Jiang BH, Liu LZ. PI3K/PTEN signaling in angiogenesis and tumorigenesis. Adv Cancer Res. 2009;102:19-65.

Oak JS, Fruman DA. Role of phosphoinositide 3-kinase signaling in autoimmunity. Autoimmunity. 2007;40:433-41.

Akanbi KA, Brodie AE, Suryawan A, Hu CY. Effect of age on the differentiation of porcine adipose stromal-vascular cells in culture. J Anim Sci. 1994;72(11):2828-35.

Kim EH, Suresh M. Role of PI3K/Akt signaling in memory CD8 T cell differentiation. Front Immunol. 2013;4:20.

Gao HW, Liu Lan, Xing DG, Liu ZH, Ren P, Li ZQ, Shan GQ, Gong MZ. NCoR negatively regulates adipogenic differentiation of mesenchymal stem cells. In Vitro Cell Dev Biol Anim. 2015;51(7):749-58.

Li B, Dong Z, Gao L, He X, Liao L, Hu C, Wang Q, Jin Y. Lipopolysaccharide differentially affects the osteogenic differentiation of periodontal ligament stem cells and bone marrow mesenchymal stem cells through Toll-like receptor 4 mediated nuclear factor κB pathway. Stem Cell Res Ther. 2014;5(3):67.

Huyck L, Ampe C, Van Troys M. The XTT cell proliferation assay applied to cell layers embedded in three-dimensional matrix. Assay Drug Dev Technol. 2012;10(4):382-92.

Son KM, Park HC, Kim NR, Lee IS, Yang HC. Enhancement of the ALP activity of C3H10T1/2 cells by the combination of an oxysterol and apatite. Biomed Mater. 2010; 5(4):044107.

Westergren-Thorsson G, Onnervik PO, Fransson LA, Malmström A. Proliferation of cultured fibroblasts is inhibited by L-iduronate-containing glycosaminoglycans. J Cell Physiol. 1991;147(3):523-30.

Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JM, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143-7.

Wang YK, Chen CS. Cell Adhesion and mechanical stimulation in the regulation of mesenchymal stem cell differentiation. Cell Mol Med. 2013;17(7):823-32.

Chen D, Ji X, Harris MA, Feng JQ, Karsenty G, Celeste AJ, Rosen V, Mundy GR, Harris SE. Differential roles for bone morphogenetic protein (BMP) receptor type IB and IA in differentiation and specification of mesenchymal precursor cells to osteoblast and adipocyte lineages. J Cell Biol. 1998;142:295-305.

Tontonoz P, Hu E, Spiegelman B. Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor gamma. Curr Opin Genet Dev. 1995;5:571-6.

Kveiborg M, Flyvbjerg A, Eriksen EF, Kassem M. 1,25-Dihydroxyvitamin D3 stimulates the production of insulin-like growth factor-binding proteins-2, -3 and -4 in human bone marrow stromal cells. Eur J Endocrinol. 2001;144(5):549-57.

Allen MJ. Biochemical markers of bone metabolism in animals: uses and limitations. Vet Clin Pathol. 2003;32:101-13.

Sila-Asna M, Bunyaratvej A, Maeda S, Kitaguchi H, Bunyaratavej N. Osteoblast differentiation and bone formation gene expression in strontiunm-inducing bone marrow mesenchymal stem cell. Kobe J Med Sci. 2007;53:25-35.

Komori T. Runx2, a multifunctional transcription factor in skeletal development. J Cell Biochem. 2002;87:1-8.

Harada H, Tagashira S, Fujiwara M, Ogawa S, Katsumata T, Yamaguchi A. Cbfa1 isoforms exert functional differences in osteoblast differentiation. J Biol Chem. 1999;274: 6972-8.

Westendorf JJ, Zaidi SK, Cascino JE, Kahler R, van Wijnen AJ. Runx2 (Cbfa 1, AML-3) interacts with histone deacetylase 6 and represses the p21 (CIP1/WAF1) promoter. Mol Cell Biol. 2002;22(22):7982-92.

Zhang C. Transcriptional regulation of bone formation by the osteoblast specific transcription factor Osx. J Orthop Surg Res. 2010;5:37.

Peng S, Zhou G, Luk KD, Cheung KM, Li Z. Strontium promotes osteogenic differentiation of mesenchymal stem cells through the Ras/MAPK signaling pathway. Cell Physiol Biochem. 2008;23:165-74.

Ghosh-Choudhury N, Abboud SL, Nishimura R, Celeste A, Mahimainathan L, Choudhury GG. Requirement of BMP-2-induced phosphatidylinositol 3-kinase and Akt serine/threonine kinase in osteoblast differentiation and Smad-dependent BMP-2 gene transcription. J Biol Chem. 2002;277:33361-8.

Kaliman P, Vinals F, Testar X, Palacin M, Zorzano A. Phosphatidylinositol 3-kinase inhibitors block differentiation of skeletal muscle cells. J Biol Chem. 1996;271:19146-51.

Sakaue H, Ogawa W, Matsumoto M, Kuroda S, Takata M, Sugimoto T, Spiegelman BM, Kasuga M. Posttranscriptional control of adipocyte differentiation through activation of phosphoinositide 3-kinase. J Biol Chem. 1998;273:28945-52.

Sugatani T, Hruska KA. Akt1/Akt2 and mammalian target of rapamycin/Bim play critical roles in osteoclast differentiation and survival, respectively, whereas Akt is dispensable for cell survival in isolated osteoclast precursors. J Biol Chem. 2005;280(5):3583-9.

Peng XD, Xu PZ, Chen ML, Hahn-Windgassen A, Skeen J, Jacobs J. Dwarfism, impaired skin development, skeletal muscle atrophy, delayed bone development, and impeded adipogenesis in mice lacking Akt1 and Akt2. Genes Dev. 2003;17:1352-65.

Alessi DR, Deak M, Casamayor A, Caudwell FB, Morrice N, Norman DG. 3-Phosphoinositide-dependent protein kinase-1 (PDK1): structural and functional homology with the Drosophila DSTPK61 kinase. Curr Biol. 1997;7(10):776-89.

Mora A, Komander D, van Aalten DM, Alessi DR. PDK1, the master regulator of AGC kinase signal transduction. Semin Cell Dev Biol. 2004;15(2):161-70.

Toker A, Newton AC. Cellular signaling: pivoting around PDK-1. Cell. 2000;103(2):185-8.

Downloads

Published

2016-09-05

How to Cite

1.
Qin Y, Cao G, Ye J, Wang P, Gao L, Wang S, Shen H. N-COR MODULATES OSTEOGENIC DIFFERENTIATION OF RAT MESENCHYMAL STEM CELLS THROUGH THE PI3K/AKT-CELL SIGNALING PATHWAY. Arch Biol Sci [Internet]. 2016Sep.5 [cited 2024Apr.16];68(3):551-9. Available from: https://www.serbiosoc.org.rs/arch/index.php/abs/article/view/973

Issue

Vol. 68 No. 3 (2016)

Section

Articles

License

Authors grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution 4.0 International License that allows others to share the work with an acknowledgment of the work’s authorship and initial publication in this journal.