Migration Of ECV304 Cells Research Articles

Endoscopic deployment of multiple plastic stents for biliary stricture after living donor liver transplantation

Objective To construct a recombinant adenovirus vector carrying hSulf-1 gene and to investigate its effect on the proliferation and migration of human umbilical vein endothelial cells.Methods The hSulf-1 protein was inserted into adenovirus genome to generate recombinant adenovirus Ad5-hSulf1,and then the virus was used to infect ECV-304 cell line.The expression of hSulf-1 protein was detected by Western blotting analysis,the cell viability was examined by MTT assay,and the cell migration ability was evaluated by wound-healing assay in ECV-304 cells.Results The recombinant adenovirus Ad5-hSulf1 was successfully constructed.Western blotting analysis showed that over-expression of hSulf1 down-regulated the phosphorylation levels of Akt and ERK in ECV-304 cells.MTT assay showed that over-expression of hSulf1 inhibited the proliferation of ECV-304 cells,with the cell survival rate decreased to(68.49±0.05)% at MOI of 50 pfu/ml and(67.78±0.06)% at MOI of 100 pfu/ml(P0.05).Wound-healing assay showed that over-expression of hSulf1 significantly inhibited the migration of ECV-304 cells compared with the control group(P0.01).Conclusion The recombinant adenovirus Ad5-hSulf1-mediated hSulf-1 over-expression can markedly inhibit the proliferation and migration of ECV-304 cells,laying a foundation for hSulf-1 related gene therapy of tumor and angiogenesis-associated diseases.

Inhibitory Effect of Paclitaxel on Endothelial Cell Adhesion and Migration

The long-term success of percutaneous coronary interventions has been limited by restenosis. Therefore, local delivery of paclitaxel, an antiproliferative agent, using drug-eluting stents has been applied to prevent in-stent restenosis. However, paclitaxel not only inhibits smooth muscle cell proliferation, but also delays re-endothelialization of the damaged site, which may cause potentially life-threatening cardiovascular adverse events, especially late and very late stent thrombosis. We investigated the role of paclitaxel in endothelial cell line ECV304 adhesion and migration. Accordingly, changes in vasodilator-stimulated phosphoprotein protein (VASP) phosphorylation and cAMP-dependent protein kinase activity during ECV304 cell detachment and reattachment were investigated as well. The results showed that the decrease in VASP phosphorylation paralleled the inhibition of cAMP-dependent protein kinase (PKA) activity in the presence of paclitaxel (10 µg/l). Cell adhesion assay and two- and three-dimensional cell migration assays were performed to determine the effect of paclitaxel on the adhesion and migration of ECV304 cells. Paclitaxel significantly suppresses the adhesion (p < 0.05) and migration of ECV304 cells (p < 0.05). These data suggest that the inhibitory effect of paclitaxel may be produced by decreasing the phosphorylation of VASP via inhibition of PKA activity during ECV304 cell adhesion and migration.

Hepatocyte growth factor protects endothelial cells against gamma ray irradiation-induced damage

To investigate the effect of HGF on proliferation, apoptosis and migratory ability of human vascular endothelial cells against gamma ray irradiation. ECV304 cells derived from adult human umbilical vein endothelial cells (HUVEC) were irradiated with a single gamma ray dose of 20 Gy. Immunocytochemistry and Western blot analysis were used to detect c-Met protein expression and HGF/c-Met signal pathway. In the HGF-treated groups, ECV304 cells were incubated with HGF (20 or 40 ng/mL) 3 h prior to irradiation. At 48 h post-irradiation, the proliferation of ECV304 cells was measured by MTT assay, the apoptosis was assessed by flow cytometry, and the migratory ability of ECV304 cells was measured by transwell chamber assay. c-Met protein is expressed in ECV304 cells and can be activated by HGF. Gamma ray irradiation inhibits proliferation and migration of ECV304 cells in a dose-dependent manner. HGF significantly promoted the proliferation of ECV304 cells, and flow cytometry revealed that HGF can inhibit apoptosis of ECV304 cells. Transwell chamber assay also showed that HGF increases migration activity of endothelial cells. HGF may afford protection to vascular endothelial cells against gamma ray irradiation-induced damage.

Effect of Sodium Ferulate on the Cell Migration Induced by TNF‐α

People's Republic of Migration of vascular cells is a crucial response to vascular injury resulting in atherosclerosis. Tumor necrosis factor‐α(TNF‐α) functions as a potent chemoattractant for cells and can change cell cytoskeleton obviously.Sodium Ferulate(SF), a principle from traditional Chinese herb medicine of Angelica, is approved by State Drugs Administration of China as a new drug for the treatment of cardiovascular diseases.This study was to employ ECV304 cells migration model induced by Tumor necrosis factor‐α(TNF‐α) and to make studies of the effect of SF on the migration of ECV304 cells .Through cell scratch wound migration assay and quantification of cell migration, we observed that TNF‐α significantly promoted cell migration of ECV304 and that SF could inhibit the migration induced by TNF‐α treated for 24h.Meanwhile the morphological changes were observed under Giemsa staining and HITACHIS‐570 scanning electron microscope.The morphologies of the migration cells treated only by TNF‐α were markedly changed:cytoplasm radial protruding,numerous pseudopods presenting filament‐like and thin strip‐like contour of some cells presenting.crab.‐ like or.hedgehog.like shape.And the morphologies of the cells treated SF and TNF‐α were similar to the control cells, exhibited regular arrangements, relatively large volume, mostly round or polygon shapes, abundant surface microvillus on the surface.These findings suggest that SF may protect cell pseudopodium and surface microvillus from TNF‐α, and inhibite the migration of ECV304 stimulated by TNF‐α, accordingly it can prevent and treat atherosclerosis.(The research is supported by national natural science foundation of China granted NO.30470680)

H-Ras Is a Negative Regulator of α3β1Integrin Expression in ECV304 Endothelial Cells

We have examined the role of Ras in integrin expression in ECV304 endothelial cells. Among the integrins examined in stable ECV304 transfectants expressing dominant active H-Ras (DAR-ECV), expression of α3β1integrin showed a prominent reduction in all the DAR-ECV clones when compared to the parental ECV304 cells. This implies that H-Ras negatively regulates the expression of α3β1integrin in ECV304 cells. When treated with inhibitors of the Ras downstream pathway (LY294002, PD98059, SB203580), the expression of α3β1integrin was up-regulated most significantly by LY294002, suggesting that among the downstream pathways of Ras, phosphatidylinositol 3-kinase is a major determinant. With the application of blocking antibody to α3β1integrin (2 – 2 × 104nM), migration of ECV304 cells was enhanced to maximal (18%) at 20 nM. These results suggest that migration of endothelial cells could be modulated by H-Ras via alteration of the expression levels of α3β1integrin.

Stimulation of endothelial cell migration in culture by ladsin, a laminin-5-like cell adhesion protein.

Ladsin is a laminin-like cell-adhesive scatter factor with potent cell motility-stimulating ability and was purified from serum-free conditioned medium of a malignant human gastric adenocarcinoma cell line STKM-1. To test its possible role in tumor angiogenesis, we investigated its effect on primary culture of endothelial cells (human umbilical vein endothelial cells) and endothelial cell line ECV304 in this study. Cell adhesion and motility effects of ladsin were observed in both types of endothelial cells. In cell-attachment assay, ladsin interacted with integrin alpha 3 beta 1 that was expressed on the endothelial cell surface. In Boyden chambers, ladsin stimulated both directed and random migration of ECV304 cells. Ladsin induced repair of artificial wounds generated in ECV304 cell monolayers by stimulating cell migration. Ladsin did not affect the growth rate of ECV304 cells at a low cell density but significantly increased the saturation cell density. These results suggest that ladsin may be involved in the adhesion and migration of endothelial cells under some physiological and pathological conditions.