Deciphering Mechanisms Controlling Placental Artery Endothelial Cell Migration Stimulated by Vascular Endothelial Growth Factor

Abstract

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.

Vascular endothelial growth factor C enhances cervical cancer migration and invasion via activation of focal adhesion kinase

“…We here found that VEGF-C increased FAK Tyr 397 phosphorylation, leading to the formation of focal adhesion complexes. Similar to our results, there is evidence showing that VEGF-C or its isoform can phosphorylate FAK and promote cell adhesion to substratum [18,19].…”

Expression and roles of Slit/Robo in human ovarian cancer

“…Cell migration was evaluated using a 24-Multiwell BD Falcon FluoroBlok Insert System (8.0 µm pores; BD Biosciences, San Jose, CA, USA) as described (Liao et al 2010a). After serum starvation for 24 h, 30,000 cells were seeded in the insert (topside of membrane) and cultured in serum-free media.…”

Rac1-Dependent Intracellular Superoxide Formation Mediates Vascular Endothelial Growth Factor–Induced Placental Angiogenesis in Vitro

“…Tube formation on matrigel was measured as previously described (5,6). Images from five microscopic fields per well were analyzed.…”

“…Endocrinology, November 2010, 151(11):5315-5325 endo.endojournals.orgare the best studied angiogenic regulatory system in the placenta (5,6,39). Compelling data have shown that VEGF and its receptors KDR and Flt1 are increased in uterine and fetal tissues accompanied with placental angiogenesis during pregnancy.…”

“…Ligand binding initiates tyrosine phosphorylation of the intracellular domains of KDR and Flt-1, resulting in activation of downstream signaling pathways (4 -6). In placental endothelial cells, we have shown that VEGF activates complex signaling pathways, including extracellular signal-regulated kinases, phosphoinositol-3-kinase/protein kinase B, and endothelial NO synthase (eNOS)/NO, etc., which are critical for endothelial cell angiogenesis and gene expression (5)(6)(7)(8)(9); however, how VEGF regulates placental angiogenesis remains incompletely understood.…”