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

Abstract

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.