Involvement of activation of C-met signaling pathway in CD151-induced HUVECs angiogenesis

Abstract

CD151 is a member of the tetraspanin family that is implicated as a promoter of pathological or physiological angiogenesis. C-Met is expressed on a variety of cells including vascular endothelial cells (VECs) and up-regulated during angiogenesis. In this study, we investigated whether CD151 regulated migration, proliferation, tube formation and angiogenesis of human umbilical VECs (HUVECs) with activation of C-Met. Moreover, we studied whether CD151 could affect the angiogenic molecules such as nitric oxide (NO), vascular cell adhesion molecule-1 (VCAM-1) and vascular endothelial growth factor (VEGF). The expression of CD151 was determined by Western blotting. The cell proliferation assay was performed using the cell counting kit-8 (CCK-8) method and cell migration was assessed in microchemotaxis chambers by using fetal bovine serum (FBS) as the chemotactic stimulus. The angiogenic molecules were evaluated using ELISA. The NO level was detected using NO detection kit. The potential involvement of various signaling pathways was explored using relevant antibodies. We found that proliferation, migration and tube formation of HUVECs were promoted by CD151 with activation of C-Met, FAK and CDC42, while they were suppressed with CD151 knockdown by RNAi. Similarly, the levels of NO, VCAM-1 and VEGF in HUVECs were increased by CD151, but they were inhibited with CD151 knockdown by RNAi. These data suggested that CD151 could promote migration, proliferation, tube formation and angiogenesis of HUVECs, which was possibly related to the C-Met signaling pathways.

Keywords

CD151 / C-Met / vascular endothelial growth factor / vascular cell adhesion molecule-1 / angiogenesis

References

[1] ChattopadhyayN, WangZ, AshmanLK, et al. . alpha3beta1 integrin-CD151, a component of the cadherin-catenin complex, regulates PTPmu expression and cell-cell adhesion. J Cell Biol, 2003, 163(6): 1351-1362 PMCID: 2173722 PMID: 14691142

[2] FitterS, TetazTJ, BerndtMC, et al. . Molecular cloning of cDNA encoding a novel platelet-endothelial cell tetraspan antigen, PETA-3. Blood, 1995, 86(4): 1348-1355 PMID: 7632941

[3] SincockPM, FitterS, PartonRG, et al. . PETA-3/CD151, a member of the transmembrane 4 superfamily, is localised to the plasma membrane and endocytic system of endothelial cells, associates with multiple integrins and modulates cell function. J Cell Sci, 1999, 112(pt6): 833-844 PMID: 10036233

[4] SincockPM, MayrhoferG, AshmanLK. Localization of the transmembrane 4 superfamily (TM4SF) member PETA-3 (CD151) in normal human tissues: comparison with CD9, CD63, and alpha5beta1 integrin. J Histochem Cytochem, 1997, 45(4): 515-525 PMID: 9111230

[5] YangXH, MirchevR, DengX, et al. . CD151 restricts the α6 integrin diffusion mode. J Cell Sci, 2012, 125(6): 1478-1487 PMCID: 3336378 PMID: 22328509

[6] NishiuchiR, SanzenN, NadaS, et al. . Potentiation of the ligand-binding activity of integrin a3β1 via association with tetraspanin CD151. Proc Natl Acad Sci USA, 2005, 102(6): 1939-1944 PMCID: 548567 PMID: 15677332

[7] ZhangXA, KazarovAR, YangX, et al. . Function of the tetraspanin CD151-alpha6beta1 integrin complex during cellular morphogenesis. Mol Biol Cell, 2002, 13(1): 1-11 PMCID: 65068 PMID: 11809818

[8] LanRF, LiuZX, LiuXC, et al. . Selective tetraspan-integrin complexes (CD81/alpha4beta1, CD151/alpha3beta1, CD-151/alpha6beta1) under conditions disrupting tetraspan interactions. Biochem J, 1999, 340(Pt1): 103-111

[9] ZhangXA, BontragerAL, HemlerME. Transmembrane-4 superfamily proteins associate with activated protein kinase C (PKC) and link PKC to specific beta(1) integrins. J Biol Chem, 2001, 276(27): 25005-25013 PMID: 11325968

[10] ZuoHJ, LinJY, LiuZY, et al. . Activation of the ERK signaling pathway is involved in CD151-induced angiogenic effects on the formation of CD151-integrin complexes. Acta Pharmacol Sin, 2010, 31(7): 805-812 PMCID: 4007724 PMID: 20581856

[11] ZhengZ, LiuZ. CD151 gene delivery increases eNOS activity and induces ECV304 migration, proliferation and tube formation. Acta Pharmacol Sin, 2007, 28(1): 66-72 PMID: 17184584

[12] NakamuraT, SakaiK, NakamuraT, et al. . Hepatocyte growth factor twenty years on: Much more than a growth factor. J Gastroenterol Hepatol, 2011, 26(Suppl1): 188-202 PMID: 21199531

[13] McKinnonH, GherardiE, ReidyM, et al. . Hepatocyte growth factor/scatter factor and MET are involved in arterial repair and atherogenesis. Am J Pathol, 2006, 168(1): 340-348 PMCID: 1592681 PMID: 16400035

[14] WiniarskiBK, WolanskaKI, RaiS, et al. . Epithelial ovarian cancer-induced angiogenic phenotype of human omental microvascular endothelial cells may occur independently of VEGF signaling. Transl Oncol, 2013, 6(6): 703-714 PMCID: 3890705 PMID: 24466373

[15] KuhlmannCR, SchaeferCA, FehseckeA, et al. . A new signaling mechanism of hepatocyte growth factor-induced endothelial proliferation. J Thromb Haemost, 2005, 3(9): 2089-2095 PMID: 16102116

[16] HongIK, JinYJ, ByunHJ, et al. . Homophilic interactions of Tetraspanin CD151 up-regulate motility and matrix metalloproteinase-9 expression of human melanoma cells through adhesion-dependent c-Jun activation signaling pathways. J Biol Chem, 2006, 281(34): 24279-24292 PMID: 16798740

[17] KohnoM, HasegawaH, MiyakeM, et al. . CD151 enhances cell motility and metastasis of cancer cells in the presence of focal adhesion kinase. Int J Cancer, 2002, 97(3): 336-343 PMID: 11774285

[18] CarmelietP. Mechanisms of angiogenesis and arteriogenesis. Nat Med, 2000, 6(4): 389-395 PMID: 10742145

[19] BuschmannI, SchaperW. Arteriogenesis versus angiogenesis: Two mechanisms of vessel growth. News Physiol Sci, 1999, 14: 121-125 PMID: 11390835

[20] ZhaoX, LuX, FengQ. Deficiency in endothelial nitric oxide synthase impairs myocardial angiogenesis. Am J Physiol Heart Circ Physiol, 2002, 283(6): H2371-H2378 PMID: 12388304

[21] MorbidelliL, DonniniS, ZicheM. Role of nitric oxide in tumor angiogenesis. Cancer Treat Res, 2004, 117: 155-167 PMID: 15015560

[22] DimmelerS, DernbachE, ZeiherAM. Phosphorylation of the endothelial nitric oxide synthase at Ser-1177 is required for VEGF-induced endothelial cell migration. FEBS Lett, 2000, 477(3): 258-262 PMID: 10908731

[23] GoochKJ, DanglerCA, FrangosJA. Exogenous, basal, and flow-induced nitric oxide production and endothelial cell proliferation. J Cell Physiol, 1997, 171(3): 252-258 PMID: 9180894

[24] MuroharaT, AsaharaT, SilverM, et al. . Nitric oxide synthase modulates angiogenesis in response to tissue ischemia. J Clin Investig, 1998, 101(11): 2567-2578 PMCID: 508846 PMID: 9616228

[25] FultonD, GrattonJP, McCabeTJ, et al. . Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature, 1999, 399(6736): 597-601 PMCID: 3637917 PMID: 10376602

[26] ConnollyDT, HeuvelmanDM, NelsonR, et al. . Tumor vascular permeability factor stimulates endothelial cell growth and angiogenesis. J Clin Invest, 1989, 84(5): 1470-1478 PMCID: 304011 PMID: 2478587

[27] LeungDW, CachianesG, KuangWJ, et al. . Vascular endothelial growth factor is a secreted angiogenic mitogen. Science, 1989, 246(4935): 1306-1309 PMID: 2479986

[28] ShuQ, LiW, LiH, et al. . Vasostatin inhibits VEGF-induced endothelial cell proliferation, tube formation and induces cell apoptosis under oxygen deprivation. Int J Mol Sci, 2014, 15(4): 6019-6030 PMCID: 4013612 PMID: 24722573

[29] FuSP, HeSY, XuB, et al. . Acupuncture promotes angiogenesis after myocardial ischemia through H3K9 acetylation regulation at VEGF gene. PLoS One, 2014, 9(4): e94604 PMCID: 3983235 PMID: 24722278

[30] DeindlE. Mechanistic insights into the functional role of VEGF and its signaling partner BDNF in angiogenic tube formation. Acta Physiol (Oxf), 2014, 211(2): 268-270

[31] PoreN, LiuS, ShuHK, et al. . Sp1 is involved in Akt-mediated induction of VEGF expression through an HIF-1-independent mechanism. Mol Biol Cell, 2004, 15(11): 4841-4853 PMCID: 524732 PMID: 15342781

[32] ZhengZ, LiuZ. CD151 gene delivery activates PI3K/Akt pathway and promotes neovascularization after myocardial infarction in rats. Mol Med, 2006, 12(9–10): 214-220 PMCID: 1770014 PMID: 17225869

[33] RudgeJS, HolashJ, HyltonD, et al. . VEGF Trap complex formation measures production rates of VEGF, providing a biomarker for predicting efficacious angiogenic blockade. Proc Natl Acad Sci U S A, 2007, 104(47): 18363-18370 PMCID: 2141784 PMID: 18000042

[34] HuoY, LeyK. Adhesion molecules and atherogenesis. Acta Physiol Scand, 2001, 173(1): 35-43 PMID: 11678724

[35] KochAE, HalloranMM, HaskellCJ, et al. . Angiogenesis mediated by soluble forms of E-selectin and vascular cell adhesion molecule-1. Nature, 1995, 376(6540): 517-519 PMID: 7543654

[36] SpringFA, GriffithsRE, MankelowTJ, et al. . Tetraspanins CD81 and CD82 facilitate α4β1-mediated adhesion of human erythroblasts to vascular cell adhesion molecule-1. PLoS One, 2013, 8(5): e62654 PMCID: 3660455 PMID: 23704882

[37] UsatyukPV, FuP, MohanV, et al. . Role of c-Met/phosphatidylinositol 3-kinase (PI3k)/Akt signaling in hepatocyte growth factor (HGF)-mediated lamellipodia formation, reactive oxygen species (ROS) generation, and motility of lung endothelial cells. J Biol Chem, 2014, 289(19): 13476-13491 PMID: 24634221

[38] SenninoB, Ishiguro-OonumaT, WeiY, et al. . Suppression of tumor invasion and metastasis by concurrent inhibition of c-Met and VEGF signaling in pancreatic neuroendocrine tumors. Cancer Discov, 2012, 2(3): 270-287 PMCID: 3354652 PMID: 22585997

[39] LuKV, ChangJP, ParachoniakCA, et al. . VEGF inhibits tumor cell invasion and mesenchymal transition through a MET/VEGFR2 complex. Cancer Cell, 2012, 22(1): 21-35 PMCID: 4068350 PMID: 22789536

[40] HanahanD, WeinbergRA. Hallmarks of cancer: the next generation. Cell, 2011, 144(5): 646-674 PMID: 21376230

[41] KlosekSK, NakashiroK, HaraS, et al. . CD151 forms a functional complex with c-Met in human salivary gland cancer cells. Biochem Biophys Res Commun, 2005, 336(2): 408-416 PMID: 16139245

[42] FrancoM, MuratoriC, CorsoS, et al. . The tetraspanin CD151 is required for Met-dependent signaling and tumor cell growth. J Biol Chem, 2010, 285(50): 38756-38764 PMCID: 2998140 PMID: 20937830