Deciphering mechanisms controlling placental artery endothelial cell migration stimulated by vascular endothelial growth factor.
Vascular endothelial growth factor (VEGF) stimulated fetoplacental artery endothelial (oFPAE) cell migration and activated multiple signaling pathways including ERK2/1, p38MAPK, Jun N-terminal kinase (JNK1/2), v-Akt murine thymoma viral oncogene homolog 1 (Akt1), and c-Src in oFPAE cells. VEGF-induced cell migration was blocked by specific kinase inhibitors of JNK1/2 (SP600125), c-Src (4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d] pyrimidine), and phosphatidylinositol 3-kinase/Akt (wortmannin) but not ERK2/1 (U0126) and p38MAPK (SB203580). VEGF-induced cell migration was associated with dynamic actin reorganization and focal adhesion as evidenced by increased stress fiber formation and phosphorylation of cofilin-1 and focal adhesion kinase (FAK) and paxillin. Inhibition of JNK1/2, c-Src, and phosphatidylinositol 3-kinase/Akt suppressed VEGF-induced stress fiber formation and cofilin-1 phosphorylation. c-Src inhibition suppressed VEGF-induced phosphorylation of focal adhesion kinase, paxillin, and focal adhesion. VEGF-induced cell migration requires endogenous nitric oxide (NO) as: 1) VEGF-stimulated phosphorylation of endothelial NO synthase (eNOS) via activation of Akt, JNK1/2, and Src; 2) a NO donor diethylenetriamine-NO-stimulated cell migration; and 3) NO synthase inhibition blocked VEGF-induced cell migration. Targeted down-regulation and overexpression of caveolin-1 both inhibited VEGF-induced cell migration. Caveolin-1 down-regulation suppressed VEGF-stimulated phosphorylation of Akt, JNK, eNOS, c-Src, and FAK; however, basal activities of c-Src and FAK were elevated in parallel with increased stress fiber formation and focal adhesion. Caveolin-1 overexpression also inhibited VEGF-induced phosphorylation of Akt, JNK, c-Src, FAK, and eNOS. Thus, VEGF-induced placental endothelial cell migration requires activation of complex pathways that are paradoxically regulated by caveolin-1.
Liao
Wu-xiang
WX
Department of Obstetrics and Gynecology, University of California-Irvine, Orange, CA 92673, USA.
Feng
Lin
L
Zheng
Jing
J
Chen
Dong-bao
DB
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D001158:Arteries / Q000166:cytology*; D002070:Butadienes / Q000494:pharmacology; D000081247:CSK Tyrosine-Protein Kinase; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D042783:Endothelial Cells / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D047468:Immunoprecipitation; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D009589:Nitrogen Oxides / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010920:Placenta / Q000098:blood supply*; D011247:Pregnancy; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D012756:Sheep; D015398:Signal Transduction / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*; D000077191:Wortmannin; D019061:src-Family Kinases
D000730:Androstadienes; D000873:Anthracenes; D002070:Butadienes; D000081247:CSK Tyrosine-Protein Kinase; D051242:Caveolin 1; D004791:Enzyme Inhibitors; D051416:Focal Adhesion Protein-Tyrosine Kinases; D007093:Imidazoles; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D052250:Nitric Oxide Synthase Type III; D009570:Nitriles; D009589:Nitrogen Oxides; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011505:Protein-Tyrosine Kinases; D011725:Pyridines; D011743:Pyrimidines; D042461:Vascular Endothelial Growth Factor A; D000077191:Wortmannin; D019061:src-Family Kinases; C412373:AG 1879; C113580:U 0126; C432165:pyrazolanthrone; C093642:SB 203580
Mol Cell Biol. 2007 Mar;27(6):2372-83
17242212
J Biol Chem. 1993 Nov 25;268(33):25009-14
7693711
J Cell Biol. 1992 Nov;119(4):893-903
1385444
Biol Reprod. 2007 Feb;76(2):259-67
17050858
EMBO J. 2006 Feb 22;25(4):713-26
16456544
Mol Cell Biol. 1994 Mar;14(3):1680-8
7509446
Proc Natl Acad Sci U S A. 2000 Sep 12;97(19):10454-9
10973481
J Cell Biol. 1968 Jun;37(3):633-49
11905197
Thromb Haemost. 2005 Sep;94(3):606-14
16268479
Biol Reprod. 2005 Mar;72(3):530-7
15564597
Am J Physiol Lung Cell Mol Physiol. 2001 Dec;281(6):L1500-11
11704547
Nat Cell Biol. 2000 Dec;2(12):E231-6
11146675
Nature. 1999 Jun 10;399(6736):601-5
10376603
Arterioscler Thromb Vasc Biol. 2003 May 1;23(5):795-801
12637337
Biol Reprod. 2001 Apr;64(4):1033-40
11259247
Mol Cell Biol. 1996 Apr;16(4):1722-33
8657148
Am J Physiol Renal Physiol. 2004 Apr;286(4):F711-9
14665433
Diabetes. 2003 Dec;52(12):2959-68
14633857
J Cell Physiol. 2008 Nov;217(2):423-32
18543249
Biol Reprod. 2006 Feb;74(2):375-82
16251502
Cell Tissue Res. 2009 Jan;335(1):249-59
18815812
Nat Med. 2000 Apr;6(4):389-95
10742145
Nat Med. 2000 Dec;6(12):1362-7
11100121
Annu Rev Biochem. 2009;78:857-902
19317650
Curr Opin Pharmacol. 2008 Aug;8(4):427-32
18625340
J Biol Chem. 1998 Jul 17;273(29):18623-32
9660836
J Soc Gynecol Investig. 1999 Jul-Aug;6(4):179-85
10486778
J Biol Chem. 1998 Mar 6;273(10):5419-22
9488658
Oncogene. 2004 Jan 8;23(1):192-200
14712224
J Cell Biol. 2007 May 21;177(4):683-94
17517963
Nat Cell Biol. 2000 May;2(5):249-56
10806474
Angiogenesis. 2004;7(4):313-21
15886875
Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13681-6
11717429
J Biol Chem. 2007 Jul 6;282(27):19619-28
17449913
J Exp Med. 2006 Dec 25;203(13):2895-906
17178917
J Clin Invest. 1998 Jun 1;101(11):2567-78
9616228
Mol Endocrinol. 2009 Sep;23(9):1428-44
19477952
Biochim Biophys Acta. 2007 May;1772(5):524-6
17320358
Cardiovasc Res. 2006 Apr 1;70(1):31-41
16540099
Nat Protoc. 2007;2(2):329-33
17406593
Nature. 2003 Jul 10;424(6945):219-23
12853963
J Biol Chem. 2006 Jun 23;281(25):17173-17179
16618703
J Biol Chem. 2004 Sep 24;279(39):40659-69
15292187
Circ Res. 2004 Jul 23;95(2):154-61
15205364
Annu Rev Cell Dev Biol. 1999;15:185-230
10611961
J Biol Chem. 1996 Jan 12;271(2):695-701
8557675
Nat Cell Biol. 2004 Feb;6(2):154-61
14743221
Biol Reprod. 1997 May;56(5):1189-97
9160718
Trends Cell Biol. 2001 Jul;11(7):288-93
11413039
Biochim Biophys Acta. 2005 Dec 30;1746(3):322-33
16019085
Circ Res. 2000 Apr 28;86(8):892-6
10785512
J Biol Chem. 2008 Apr 11;283(15):10089-96
18252715
FEBS Lett. 1995 May 8;364(2):229-33
7750577
EMBO J. 2006 Nov 1;25(21):5058-70
17053785
J Biol Chem. 2005 Jun 3;280(22):20995-1003
15790570
Circ Res. 2007 Mar 30;100(6):782-94
17395884
J Cell Sci. 2004 Sep 15;117(Pt 20):4619-28
15371522
J Biol Chem. 2002 Feb 8;277(6):4277-84
11729179
Circ Res. 2005 Apr 1;96(6):635-42
15731459
J Biol Chem. 2004 Nov 26;279(48):50446-54
15371454
J Biol Chem. 2005 Feb 4;280(5):3541-7
15504729
Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5192-6
1594631
Placenta. 2008 Aug;29(8):708-17
18571718
Cell Calcium. 2009 Jun;45(6):625-33
19324409
Nat Med. 2003 Jun;9(6):669-76
12778165
Oncogene. 2004 Feb 12;23(6):1275-82
14676843
Proc Natl Acad Sci U S A. 2005 Jan 4;102(1):204-9
15615855
Biol Reprod. 2008 Jan;78(1):143-50
17901071
Mol Biol Cell. 2003 Jan;14(1):334-47
12529448
J Biol Chem. 2007 Mar 9;282(10):7232-41
17213184
Oncogene. 1997 Oct;15(18):2169-77
9393975
Mol Biol Cell. 2007 Nov;18(11):4659-68
17855507
J Biol Chem. 2003 Feb 7;278(6):3713-9
12446716
Endocrinology. 1999 Mar;140(3):1399-407
10067868
J Clin Invest. 2003 Mar;111(5):649-58
12618519
Nature. 1999 Jun 10;399(6736):597-601
10376602
Mol Biol Cell. 2003 Aug;14(8):3156-68
12925753
FASEB J. 2004 Dec;18(15):1801-11
15576483
Am J Pathol. 2002 Apr;160(4):1405-23
11943725
J Biol Chem. 2001 Dec 28;276(52):49289-98
11682481
Exp Cell Res. 2007 Jan 1;313(1):121-32
17055484
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2604-9
11226286
Biochem Biophys Res Commun. 1996 Apr 25;221(3):670-4
8630019
Nat Cell Biol. 2008 Mar;10(3):329-37
18264090