Home Life Sciences Overexpression of miR-126 promotes the differentiation of mesenchymal stem cells toward endothelial cells via activation of PI3K/Akt and MAPK/ERK pathways and release of paracrine factors
Article
Licensed
Unlicensed Requires Authentication

Overexpression of miR-126 promotes the differentiation of mesenchymal stem cells toward endothelial cells via activation of PI3K/Akt and MAPK/ERK pathways and release of paracrine factors

  • Feng Huang , Zhen-fei Fang , Xin-qun Hu , Liang Tang , Sheng-hua Zhou EMAIL logo and Jian-ping Huang EMAIL logo
Published/Copyright: May 29, 2013

Abstract

The endothelial cell (EC)-specific miRNA, miR-126, is known to promote angiogenesis in response to angiogenic factors by repressing negative regulators of signal transduction pathways; however, whether miR-126 might regulate the differentiation of stem cells toward endothelial lineage remains unknown. To answer this question, in this study mesenchymal stem cells (MSCs) harvested from C57BL/6 mouse bone marrow were transfected with miR-126 (MSCmiR-126) using recombinant lentiviral vectors. Results showed the para-secretion and the expression levels of phosphorylated PI3K p85, Akt, p38, ERK1 protein in the MSCmiR-126 group were dramatically increased when compared with the control group. With half culture medium refreshed every 3 days, a small number of 6-day-cultured MSCmiR-126 differentiated into endothelial-like cells and most of 9-day-cultured MSCmiR-126 formed a cobblestone-like structure. These differentiated cells evidently expressed EC-specific makers and possessed mature ECs function, while inhibition of paracrine factors suppressed the MSC-EC differentiation. Strikingly, the increased secretion of MSCmiR-126 and their endothelial-differentiated potential were deprived by using a PI3K or MEK chemical inhibitor. Our results suggest that overexpression of miR-126 agumenting the endothelial differentiation of MSCs might in part be attributable to the activation of PI3K/Akt and MAPK/ERK pathways and an increased release of paracrine factors.


Corresponding authors: Sheng-hua Zhou, Department of Cardiology, Second Xiangya Hospital of Central South University, Changsha 410011, Hunan, China; and Jian-ping Huang, Alibaba Business College, Hangzhou Normal University, Hangzhou 310036, Zhejiang, China

This work was supported by 2009 National Nature Scientific Funding of China (No. 30871053).

References

Bartel, D.P. (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297.10.1016/S0092-8674(04)00045-5Search in Google Scholar PubMed

Chen, J.J. and Zhou, S.H. (2011). Mesenchymal stem cells overexpressing MiR-126 enhance ischemic angiogenesis via the AKT/ERK-related pathway. Cardiol. J. 18, 675–681.10.5603/CJ.2011.0032Search in Google Scholar

Dimmeler, S., Aicher, A., Vasa, M., Mildner-Rihm, C., Adler, K., Tiemann, M., Rutten, H., Fichtlscherer, S., Martin, H., and Zeiher, A.M. (2001). HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J. Clin. Invest. 108, 391–397.10.1172/JCI200113152Search in Google Scholar

Evseenko, D., Zhu, Y., Schenke-Layland, K., Kuo, J., Latour, B., Ge, S., Scholes, J., Dravid, G., Li, X., MacLellan, W.R., et al. (2010). Mapping the first stages of mesoderm commitment during differentiation of human embryonic stem cells. Proc. Natl. Acad. Sci. USA 107, 13742–13747.10.1073/pnas.1002077107Search in Google Scholar PubMed PubMed Central

Fish, J.E., Santoro, M.M., Morton, S.U., Yu, S., Yeh, R.F., Wythe, J.D., Ivey, K.N., Bruneau, B.G., Stainier, D.Y., and Srivastava, D. (2008). miR-126 regulates angiogenic signaling and vascular integrity. Dev. Cell 15, 272–284.10.1016/j.devcel.2008.07.008Search in Google Scholar PubMed PubMed Central

Gnecchi, M., He, H., Noiseux, N., Liang, O.D., Zhang, L., Morello, F., Mu, H., Melo, L.G., Pratt, R.E., Ingwall, J.S., et al. (2006). Evidence supporting paracrine hypothesis for Akt-modified mesenchymal stem cell-mediated cardiac protection and functional improvement. FASEB J. 20, 661–669.10.1096/fj.05-5211comSearch in Google Scholar PubMed

Gnecchi, M., Zhang, Z., Ni, A., and Dzau, V.J. (2008). Paracrine mechanisms in adult stem cell signaling and therapy. Circ. Res. 103, 1204–1219.10.1161/CIRCRESAHA.108.176826Search in Google Scholar PubMed PubMed Central

Han, J., Lee, Y., Yeom, K.H., Kim, Y.K., Jin, H., and Kim, V.N. (2004). The Drosha-DGCR8 complex in primary microRNA processing. Genes Dev. 18, 3016–3027.10.1101/gad.1262504Search in Google Scholar PubMed PubMed Central

He, L. and Hannon, G.J. (2004). MicroRNAs: small RNAs with a big role in gene regulation. Nat. Rev. Genet. 5, 522–531.10.1038/nrg1379Search in Google Scholar PubMed

Ivey, K.N., Muth, A., Arnold, J., King, F.W., Yeh, R.F., Fish, J.E., Hsiao, E.C., Schwartz, R.J., Conklin, B.R., Bernstein, H.S., et al. (2008). MicroRNA regulation of cell lineages in mouse and human embryonic stem cells. Cell Stem Cell 2, 219–229.10.1016/j.stem.2008.01.016Search in Google Scholar PubMed PubMed Central

Kane, N.M., Meloni, M., Spencer, H.L., Craig, M.A., Strehl, R., Milligan, G., Houslay, M.D., Mountford, J.C., Emanueli, C., and Baker, A.H. (2010). Derivation of endothelial cells from human embryonic stem cells by directed differentiation: analysis of microRNA and angiogenesis in vitro and in vivo. Arterioscler. Thromb. Vasc. Biol. 30, 1389–1397.10.1161/ATVBAHA.110.204800Search in Google Scholar PubMed

Kane, N.M., Howard, L., Descamps, B., Meloni, M., McClure, J., Lu, R., McCahill, A., Breen, C., Mackenzie, R.M., Delles, C., et al. (2012). Role of microRNAs 99b, 181a, and 181b in the differentiation of human embryonic stem cells to vascular endothelial cells. Stem Cells 30, 643–654.10.1002/stem.1026Search in Google Scholar PubMed PubMed Central

Kinnaird, T., Stabile, E., Burnett, M.S., Lee, C.W., Barr, S., Fuchs, S., and Epstein, S.E. (2004). Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. Circ. Res. 94, 678–685.10.1161/01.RES.0000118601.37875.ACSearch in Google Scholar PubMed

Krampera, M., Pasini, A., Rigo, A., Scupoli, M.T., Tecchio, C., Malpeli, G., Scarpa, A., Dazzi, F., Pizzolo, G., and Vinante, F. (2005). HB-EGF/HER-1 signaling in bone marrow mesenchymal stem cells: inducing cell expansion and reversibly preventing multilineage differentiation. Blood 106, 59–66.10.1182/blood-2004-09-3645Search in Google Scholar PubMed

Lee, Y., Ahn, C., Han, J., Choi, H., Kim, J., Yim, J., Lee, J., Provost, P., Radmark, O., Kim, S., et al. (2003). The nuclear RNase III Drosha initiates microRNA processing. Nature 425, 415–419.10.1038/nature01957Search in Google Scholar PubMed

Lee, Y., Kim, M., Han, J., Yeom, K.H., Lee, S., Baek, S.H., and Kim, V.N. (2004). MicroRNA genes are transcribed by RNA polymerase II. EMBO J. 23, 4051–4060.10.1038/sj.emboj.7600385Search in Google Scholar PubMed PubMed Central

Li, H., Zuo, S., He, Z., Yang, Y., Pasha, Z., Wang, Y., and Xu, M. (2010). Paracrine factors released by GATA-4 overexpressed mesenchymal stem cells increase angiogenesis and cell survival. Am. J. Physiol. Heart Circ. Physiol. 299, H1772–1781.10.1152/ajpheart.00557.2010Search in Google Scholar PubMed PubMed Central

Lim, S.Y., Kim, Y.S., Ahn, Y., Jeong, M.H., Hong, M.H., Joo, S.Y., Nam, K.I., Cho, J.G., Kang, P.M., and Park, J.C. (2006). The effects of mesenchymal stem cells transduced with Akt in a porcine myocardial infarction model. Cardiovasc. Res. 70, 530–542.10.1016/j.cardiores.2006.02.016Search in Google Scholar PubMed

Ohtani, K. and Dimmeler, S. (2011). Control of cardiovascular differentiation by microRNAs. Basic Res. Cardiol. 106, 5–11.10.1007/s00395-010-0139-7Search in Google Scholar PubMed

Small, E.M. and Olson, E.N. (2011). Pervasive roles of microRNAs in cardiovascular biology. Nature 469, 336–342.10.1038/nature09783Search in Google Scholar PubMed PubMed Central

Takaya, T., Ono, K., Kawamura, T., Takanabe, R., Kaichi, S., Morimoto, T., Wada, H., Kita, T., Shimatsu, A., and Hasegawa, K. (2009). MicroRNA-1 and MicroRNA-133 in spontaneous myocardial differentiation of mouse embryonic stem cells. Circ. J. 73, 1492–1497.10.1253/circj.CJ-08-1032Search in Google Scholar

Tiscornia, G. and Izpisua Belmonte, J.C. (2010). MicroRNAs in embryonic stem cell function and fate. Genes Dev. 24, 2732–2741.10.1101/gad.1982910Search in Google Scholar PubMed PubMed Central

Urbich, C., Aicher, A., Heeschen, C., Dernbach, E., Hofmann, W.K., Zeiher, A.M., and Dimmeler, S. (2005). Soluble factors released by endothelial progenitor cells promote migration of endothelial cells and cardiac resident progenitor cells. J. Mol. Cell. Cardiol. 39, 733–742.10.1016/j.yjmcc.2005.07.003Search in Google Scholar PubMed

Urbich, C., Kuehbacher, A., and Dimmeler, S. (2008). Role of microRNAs in vascular diseases, inflammation, and angiogenesis. Cardiovasc. Res. 79, 581–588.10.1093/cvr/cvn156Search in Google Scholar PubMed

Wang, S., Aurora, A.B., Johnson, B.A., Qi, X., McAnally, J., Hill, J.A., Richardson, J.A., Bassel-Duby, R., and Olson, E.N. (2008). The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev. Cell 15, 261–271.10.1016/j.devcel.2008.07.002Search in Google Scholar PubMed PubMed Central

Wang, H., Cao, F., De, A., Cao, Y., Contag, C., Gambhir, S.S., Wu, J.C., and Chen, X. (2009). Trafficking mesenchymal stem cell engraftment and differentiation in tumor-bearing mice by bioluminescence imaging. Stem Cells 27, 1548–1558.10.1002/stem.81Search in Google Scholar PubMed PubMed Central

Wu, F., Yang, Z., and Li, G. (2009). Role of specific microRNAs for endothelial function and angiogenesis. Biochem. Biophys. Res. Commun. 386, 549–553.10.1016/j.bbrc.2009.06.075Search in Google Scholar PubMed PubMed Central

Xu, J., Liu, X., Jiang, Y., Chu, L., Hao, H., Liua, Z., Verfaillie, C., Zweier, J., Gupta, K., and Liu, Z. (2008). MAPK/ERK signalling mediates VEGF-induced bone marrow stem cell differentiation into endothelial cell. J. Cell. Mol. Med. 12, 2395–2406.10.1111/j.1582-4934.2008.00266.xSearch in Google Scholar PubMed PubMed Central

Zou, J., Li, W.Q., Li, Q., Li, X.Q., Zhang, J.T., Liu, G.Q., Chen, J., Qiu, X.X., Tian, F.J., Wang, Z.Z., et al. (2011). Two functional microRNA-126s repress a novel target gene p21-activated kinase 1 to regulate vascular integrity in zebrafish. Circ. Res. 108, 201–209.10.1161/CIRCRESAHA.110.225045Search in Google Scholar PubMed

Received: 2013-1-11
Accepted: 2013-5-21
Published Online: 2013-05-29
Published in Print: 2013-09-01

©2013 by Walter de Gruyter Berlin Boston

Articles in the same Issue

  1. Masthead
  2. Masthead
  3. Reviews
  4. Imaging the invisible: resolving cellular microcompartments by superresolution microscopy techniques
  5. Structure and function of MK5/PRAK: the loner among the mitogen-activated protein kinase-activated protein kinases
  6. Functional ribosome biogenesis is a prerequisite for p53 destabilization: impact of chemotherapy on nucleolar functions and RNA metabolism
  7. Interleukin-6 and interleukin-11: same same but different
  8. Cathepsin K: a unique collagenolytic cysteine peptidase
  9. Research Articles/Short Communications
  10. Protein Structure and Function
  11. The active form of goat insulin-like peptide 3 (INSL3) is a single-chain structure comprising three domains B-C-A, constitutively expressed and secreted by testicular Leydig cells
  12. Molecular Medicine
  13. Zinc-dependent contact system activation induces vascular leakage and hypotension in rodents
  14. Cell Biology and Signaling
  15. ACE inhibition enhances bradykinin relaxations through nitric oxide and B1 receptor activation in bovine coronary arteries
  16. Changes in COX-2 and oxidative damage factors during differentiation of human mesenchymal stem cells to hepatocyte-like cells is associated with downregulation of P53 gene
  17. Overexpression of miR-126 promotes the differentiation of mesenchymal stem cells toward endothelial cells via activation of PI3K/Akt and MAPK/ERK pathways and release of paracrine factors
  18. Novel Techniques
  19. Synthesis of a novel benzodifuran derivative and its molecular recognition of poly rA RNA
Downloaded on 15.1.2026 from https://www.degruyterbrill.com/document/doi/10.1515/hsz-2013-0107/html
Scroll to top button