| Hepatocellular carcinoma (HCC) is one of the common malignant tumors in the world, of which the mortality rate ranked second in our country. Growth and metastasis of most malignant solid tumors including HCC depend on angiogenesis which could provide the proliferation of tumor cell with nutrition and metabolism pathway, promote the growth and metastasis of tumor. So, to study the mechanism on angiogenesis may be supply a target for treatment of HCC.Objective Vascular endothelial cadherin (VE-cadherin) is a class of cell adhesion molecule, which specificly express in vascular endothelial cells and is dispensable in the process of angiogenesis. The purpose of this study is to investigate the role of VE-cadherin on the angiogenesis of HCC (hepatocellular carcinoma), the association with vascular endothelial grow factor(VEGF165), and to provide a new target and experimental basis for HCC (hepatocellular carcinoma) anti-angiogenic therapy.Methods The 22 samples of hepatocellular carcinoma (HCC) were taken to confirm the VE-Cadherin and VEGF165 expressions by immunohistochemistry. The expression of VEGF165 in the supernatant from HepG2 cell culture was detected by ELISA, After HepG2 cell was stimulated by VEGF165, the expression of VE-Cadherin was detected by RT-PCR. HepG2 cellculture supernatants were collected and used to stimulate HUVECs cells. The activation of MAPK and VE-cadherin Y658 site was detected by Western blot. Trans-well assay was used to detect the migrational ability of endothelial cell. Effect of VE-cadherin downregulation in hepatocellular carcinoma (HCC) cells supernatant on the proliferation, cell ycle and apoptosis of human umbilical vein endothelial cells(HUVECs):After 72 h transfected, HepG2 cells supernatants in each group were collected, the expression of VE-cadherin in transiently transfected hepatocellular carcinoma (HCC) cells was detected by RT-PCR and Western blot; and the effect of supernatant on the proliferation was detected by MTS assay; the cell cycle and apoptosis were detected by FCM, and matrigel tubule formation of HUVECs were analyzed.Results Compared with adjacent tissue, the expressions of VE-Cadherin and VEGF165 in liver cancer was significantly higher, and have a certain correlation with disease progression. The expressions of VE-Cadherin in HepG2 cell can gradually increase along with the VEGF165 concentration, which may imply synergy between VE-adherin and VEGF165 in angiogenesis of HCC. Using HepG2 cell culture supernatant stimulated HUVECS endothelial cells, we found the MAPK and the phosphorylation of VE-cadherin on Y658 were activation, and the rate of migration in endothelial cells was also enhanced. After downregulation the expression of VE-Cadherin in HepG2 cell by siRNA, The proliferation of HUVECs in the interference group supernatant was inhibited significantly, the inhibition rates of 24h,48h and 72h were 38.2%,45.8%and 39.1%, respectively; After 48h, the apoptosis rate of HUVECs in the interference group supernatant was(33.02±0.25)%, which was significantly higher than negative control group (13.71±1.29)%(P<0.05); The tube formation of HUVECs in the interference group supernatant was 1.65±0.80, which was significantly lower than negative control group (15.78±2.65) (P<0.05).Conclusion VE-cadherin and VEGFk,5 are highly expressed in hepatocellular carcinoma and vascular endothelial cells, and their expression are positively correlated, suggesting that VE-cadherin may play an important role in angiogenesis in hepatocellular carcinoma, and may have a synergistic effect with VEGF165.VE-cadherin secreted by HepG2 cells have the effect of promote proliferation and anti-apoptoticon to HUVECs, but have no effect on cell cycle, indicating that VE-cadherin may have had an impact on the biological behavior of HUVECs; VE-cadherin secreted by HepG2 cells can promote the tube formation of HUVECs, indicating that VE-cadherin involved in the occurrence and development of hepatocellular carcinoma by promoting angiogenesis. |
PDF Full Text Request