Growth arrest-specific gene 6 protein promotes the proliferation and migration of endothelial progenitor cells through the PI3K/AKT signaling pathway

  • Authors:
    • Pei-Yuan Zuo
    • Xing-Lin Chen
    • Ying-Hong Lei
    • Cheng-Yun Liu
    • Yu-Wei Liu
  • View Affiliations

  • Published online on: April 24, 2014     https://doi.org/10.3892/ijmm.2014.1754
  • Pages: 299-306
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Abstract

Endothelial progenitor cells (EPCs) play an important role in endothelial repair and vascular regeneration. Growth arrest-specific gene 6 (Gas6) is a novel key regulator of the vascular system, which is linked to a number of cardiovascular diseases. However, the effects of Gas6 on EPCs have not been elucidated to date. The present study was designed to determine the biological function of EPCs treated with Gas6 and to eludicate the underlying mechanisms. EPCs were isolated from umbilical cord blood and treated with various concentrations (25, 50, 100 and 200 ng/ml) of Gas6. The proliferation, migration and angiogenesis of the Gas6-treated EPCs were evaluated by MTT assay, Transwell assay and in vitro tube formation assay, respectively. The phosphorylation status of AKT and ERK was evaluated by western blot analysis. The results demonstrated that treatment with Gas6 enhanced the proliferation and migration of the EPCs in a dose-dependent manner. However, Gas6 did not promote the differentiation of EPCs on Matrigel. Gas6 induced the phosphorylation of AKT, but not that of ERK. The enhanced proliferation and migration induced by Gas6 was markedly suppressed by the inhibitor of PI3K but not by that of ERK. These results suggest that Gas6 activates the AKT signaling pathway, which, in turn, promotes the proliferation and migration of EPCs.

References

1 

Walter DH, Rittig K, Bahlmann FH, et al: Statin therapy accelerates reendothelialization: a novel effect involving mobilization and incorporation of bone marrow-derived endothelial progenitor cells. Circulation. 105:3017–3024. 2002. View Article : Google Scholar

2 

He T, Smith LA, Harrington S, Nath KA, Caplice NM and Katusic ZS: Transplantation of circulating endothelial progenitor cells restores endothelial function of denuded rabbit carotid arteries. Stroke. 35:2378–2384. 2004. View Article : Google Scholar : PubMed/NCBI

3 

Werner N, Junk S, Laufs U, et al: Intravenous transfusion of endothelial progenitor cells reduces neointima formation after vascular injury. Circ Res. 93:e17–e24. 2003. View Article : Google Scholar : PubMed/NCBI

4 

Li DW, Liu ZQ, Wei J, Liu Y and Hu LS: Contribution of endothelial progenitor cells to neovascularization (Review). Int J Mol Med. 30:1000–1006. 2012.PubMed/NCBI

5 

Hafizi S and Dahlback B: Gas6 and protein S. Vitamin K-dependent ligands for the Axl receptor tyrosine kinase subfamily. FEBS J. 273:5231–5244. 2006.PubMed/NCBI

6 

Stitt TN, Conn G, Gore M, et al: The anticoagulation factor protein S and its relative, Gas6, are ligands for the Tyro 3/Axl family of receptor tyrosine kinases. Cell. 80:661–670. 1995. View Article : Google Scholar : PubMed/NCBI

7 

Schneider C, King RM and Philipson L: Genes specifically expressed at growth arrest of mammalian cells. Cell. 54:787–793. 1988. View Article : Google Scholar : PubMed/NCBI

8 

Melaragno MG, Cavet ME, Yan C, et al: Gas6 inhibits apoptosis in vascular smooth muscle: role of Axl kinase and Akt. J Mol Cell Cardiol. 37:881–887. 2004. View Article : Google Scholar : PubMed/NCBI

9 

Stenhoff J, Dahlback B and Hafizi S: Vitamin K-dependent Gas6 activates ERK kinase and stimulates growth of cardiac fibroblasts. Biochem Biophys Res Commun. 319:871–878. 2004. View Article : Google Scholar : PubMed/NCBI

10 

McCloskey P, Fridell YW, Attar E, et al: GAS6 mediates adhesion of cells expressing the receptor tyrosine kinase Axl. J Biol Chem. 272:23285–23291. 1997. View Article : Google Scholar : PubMed/NCBI

11 

Nakano T, Higashino K, Kikuchi N, et al: Vascular smooth muscle cell-derived, Gla-containing growth-potentiating factor for Ca(2+)-mobilizing growth factors. J Biol Chem. 270:5702–5705. 1995.PubMed/NCBI

12 

Melaragno MG, Wuthrich DA, Poppa V, et al: Increased expression of Axl tyrosine kinase after vascular injury and regulation by G protein-coupled receptor agonists in rats. Circ Res. 83:697–704. 1998. View Article : Google Scholar : PubMed/NCBI

13 

O’Donnell K, Harkes IC, Dougherty L and Wicks IP: Expression of receptor tyrosine kinase Axl and its ligand Gas6 in rheumatoid arthritis: evidence for a novel endothelial cell survival pathway. Am J Pathol. 154:1171–1180. 1999.PubMed/NCBI

14 

Hasanbasic I, Cuerquis J, Varnum B and Blostein MD: Intracellular signaling pathways involved in Gas6-Axl-mediated survival of endothelial cells. Am J Physiol Heart Circ Physiol. 287:H1207–H1213. 2004. View Article : Google Scholar : PubMed/NCBI

15 

D’Arcangelo D, Gaetano C and Capogrossi MC: Acidification prevents endothelial cell apoptosis by Axl activation. Circ Res. 91:e4–e12. 2002.PubMed/NCBI

16 

Hasanbasic I, Rajotte I and Blostein M: The role of gamma-carboxylation in the anti-apoptotic function of gas6. J Thromb Haemost. 3:2790–2797. 2005. View Article : Google Scholar : PubMed/NCBI

17 

Ruan GX and Kazlauskas A: Axl is essential for VEGF-A-dependent activation of PI3K/Akt. Embo J. 31:1692–1703. 2012. View Article : Google Scholar : PubMed/NCBI

18 

Alciato F, Sainaghi PP, Sola D, Castello L and Avanzi GC: TNF-alpha, IL-6, and IL-1 expression is inhibited by GAS6 in monocytes/macrophages. J Leukoc Biol. 87:869–875. 2010. View Article : Google Scholar : PubMed/NCBI

19 

Ganopolsky JG, Abid MR, Aird WC and Blostein MD: GAS6-induced signaling in human endothelial cells is mediated by FOXO1a. J Thromb Haemost. 6:1804–1811. 2008. View Article : Google Scholar : PubMed/NCBI

20 

Kawasaki K, Watabe T, Sase H, et al: Ras signaling directs endothelial specification of VEGFR2(+) vascular progenitor cells. J Cell Biol. 181:131–141. 2008.PubMed/NCBI

21 

Xu J, Liu X, Jiang Y, et al: MAPK/ERK signalling mediates VEGF-induced bone marrow stem cell differentiation into endothelial cell. J Cell Mol Med. 12:2395–2406. 2008. View Article : Google Scholar : PubMed/NCBI

22 

Yan X, Cai S, Xiong X, et al: Chemokine receptor CXCR7 mediates human endothelial progenitor cells survival, angiogenesis, but not proliferation. J Cell Biochem. 113:1437–1446. 2012. View Article : Google Scholar : PubMed/NCBI

23 

Wang F, Wang Y, Zhang L and Zou L: Gene modification with integrin-linked kinase improves function of endothelial progenitor cells in pre-eclampsia in vitro. J Cell Biochem. 112:3103–3111. 2011. View Article : Google Scholar : PubMed/NCBI

24 

Kalka C, Masuda H, Takahashi T, et al: Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci USA. 97:3422–3427. 2000. View Article : Google Scholar : PubMed/NCBI

25 

Kawamoto A, Tkebuchava T, Yamaguchi J, et al: Intramyocardial transplantation of autologous endothelial progenitor cells for therapeutic neovascularization of myocardial ischemia. Circulation. 107:461–468. 2003. View Article : Google Scholar

26 

Nakano T, Kawamoto K, Kishino J, Nomura K, Higashino K and Arita H: Requirement of gamma-carboxyglutamic acid residues for the biological activity of Gas6: contribution of endogenous Gas6 to the proliferation of vascular smooth muscle cells. Biochem J. 323:387–392. 1997.PubMed/NCBI

27 

Yanagita M, Arai H, Nakano T, et al: Gas6 induces mesangial cell proliferation via latent transcription factor STAT3. J Biol Chem. 276:42364–42369. 2001. View Article : Google Scholar : PubMed/NCBI

28 

Bellosta P, Zhang Q, Goff SP and Basilico C: Signaling through the ARK tyrosine kinase receptor protects from apoptosis in the absence of growth stimulation. Oncogene. 15:2387–2397. 1997. View Article : Google Scholar : PubMed/NCBI

29 

Goruppi S, Ruaro E, Varnum B and Schneider C: Gas6-mediated survival in NIH3T3 cells activates stress signalling cascade and is independent of Ras. Oncogene. 18:4224–4236. 1999. View Article : Google Scholar : PubMed/NCBI

30 

Allen MP, Linseman DA, Udo H, et al: Novel mechanism for gonadotropin-releasing hormone neuronal migration involving Gas6/Ark signaling to p38 mitogen-activated protein kinase. Mol Cell Biol. 22:599–613. 2002. View Article : Google Scholar : PubMed/NCBI

31 

Fridell YW, Villa JJ, Attar EC and Liu ET: GAS6 induces Axl-mediated chemotaxis of vascular smooth muscle cells. J Biol Chem. 273:7123–7126. 1998. View Article : Google Scholar : PubMed/NCBI

32 

Angelillo-Scherrer A, de Frutos P, Aparicio C, et al: Deficiency or inhibition of Gas6 causes platelet dysfunction and protects mice against thrombosis. Nat Med. 7:215–221. 2001. View Article : Google Scholar : PubMed/NCBI

33 

Jiang L, Liu CY, Yang QF, Wang P and Zhang W: Plasma level of growth arrest-specific 6 (GAS6) protein and genetic variations in the GAS6 gene in patients with acute coronary syndrome. Am J Clin Pathol. 131:738–743. 2009. View Article : Google Scholar : PubMed/NCBI

34 

Holland SJ, Powell MJ, Franci C, et al: Multiple roles for the receptor tyrosine kinase axl in tumor formation. Cancer Res. 65:9294–9303. 2005. View Article : Google Scholar : PubMed/NCBI

35 

Gallicchio M, Mitola S, Valdembri D, et al: Inhibition of vascular endothelial growth factor receptor 2-mediated endothelial cell activation by Axl tyrosine kinase receptor. Blood. 105:1970–1976. 2005. View Article : Google Scholar : PubMed/NCBI

36 

Li Y, Ye X, Tan C, et al: Axl as a potential therapeutic target in cancer: role of Axl in tumor growth, metastasis and angiogenesis. Oncogene. 28:3442–3455. 2009. View Article : Google Scholar : PubMed/NCBI

37 

Ruan GX and Kazlauskas A: Lactate engages receptor tyrosine kinases axl, tie2, and vascular endothelial growth factor receptor 2 to activate phosphoinositide 3-kinase/akt and promote angiogenesis. J Biol Chem. 288:21161–21172. 2013. View Article : Google Scholar

38 

Allen MP, Zeng C, Schneider K, et al: Growth arrest-specific gene 6 (Gas6)/adhesion related kinase (Ark) signaling promotes gonadotropin-releasing hormone neuronal survival via extracellular signal-regulated kinase (ERK) and Akt. Mol Endocrinol. 13:191–201. 1999. View Article : Google Scholar

39 

Shankar SL, O’Guin K, Cammer M, et al: The growth arrest-specific gene product Gas6 promotes the survival of human oligodendrocytes via a phosphatidylinositol 3-kinase-dependent pathway. J Neurosci. 23:4208–4218. 2003.PubMed/NCBI

40 

Wang JY, Lee YT, Chang PF and Chau LY: Hemin promotes proliferation and differentiation of endothelial progenitor cells via activation of AKT and ERK. J Cell Physiol. 219:617–625. 2009. View Article : Google Scholar : PubMed/NCBI

41 

Munoz-Chapuli R, Quesada AR and Angel MM: Angiogenesis and signal transduction in endothelial cells. Cell Mol Life Sci. 61:2224–2243. 2004. View Article : Google Scholar : PubMed/NCBI

42 

Park IK, Giovenzana C, Hughes TL, Yu J, Trotta R and Caligiuri MA: The Axl/Gas6 pathway is required for optimal cytokine signaling during human natural killer cell development. Blood. 113:2470–2477. 2009. View Article : Google Scholar : PubMed/NCBI

43 

Angelillo-Scherrer A, Burnier L, Lambrechts D, et al: Role of Gas6 in erythropoiesis and anemia in mice. J Clin Invest. 118:583–596. 2008.PubMed/NCBI

44 

Chao H and Hirschi KK: Hemato-vascular origins of endothelial progenitor cells? Microvasc Res. 79:169–173. 2010. View Article : Google Scholar : PubMed/NCBI

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July 2014
Volume 34 Issue 1

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APA
Zuo, P., Chen, X., Lei, Y., Liu, C., & Liu, Y. (2014). Growth arrest-specific gene 6 protein promotes the proliferation and migration of endothelial progenitor cells through the PI3K/AKT signaling pathway. International Journal of Molecular Medicine, 34, 299-306. https://doi.org/10.3892/ijmm.2014.1754
MLA
Zuo, P., Chen, X., Lei, Y., Liu, C., Liu, Y."Growth arrest-specific gene 6 protein promotes the proliferation and migration of endothelial progenitor cells through the PI3K/AKT signaling pathway". International Journal of Molecular Medicine 34.1 (2014): 299-306.
Chicago
Zuo, P., Chen, X., Lei, Y., Liu, C., Liu, Y."Growth arrest-specific gene 6 protein promotes the proliferation and migration of endothelial progenitor cells through the PI3K/AKT signaling pathway". International Journal of Molecular Medicine 34, no. 1 (2014): 299-306. https://doi.org/10.3892/ijmm.2014.1754