Construction of basic fibroblast growth factor adenovirus vector and its expression in human umbilical vein endothelial cells
10.3724/SP.J.1008.2008.00408
- Author:
Ling-Li GUO
1
Author Information
1. Department of Plastic Surgery
- Publication Type:Journal Article
- Keywords:
Adenoviriadae;
Basic fibroblast growth factor 2;
Endothelium;
Gene expression;
Umbilical veins;
Vascular
- From:
Academic Journal of Second Military Medical University
2010;28(4):408-411
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct an adenovirus vector harboring human basic fibroblast growth factor (bFGF) cDNA and investigate the expression of bFGF in human umbilical vein endothelial cells (HUVEC) in vitro. Methods: The adenovirus expression vector Ad5-bFGF was constructed by homologous recombination technique. The best value of MOI was tested by transfecting human umbilical vein endothelial cells (HUVEC) with Ad5-GFP. Ad5-bFGF was used to transfect HUVEC at the obtained value of MOI and the expression of bFGF protein was detected by immunocytochemistry method and Western blotting. Results: The best value of MOI for adenovirus 5 to transfect HUVEC was 200 and the transfection rate was 90%. Immunocytochemistry method and Western blotting showed that bFGF was expressed in HUVEC after transfection with Ad5-bFGF and the expression was significantly higher than that in untransfected HUVEC (P < 0.05). Conclusion: We have successfully constructed a recombined adenovirus vector Ad5-bFGF which can be expressed in vitro in HUVEC, paving a way for application of bFGF in gene therapy and study of tumorigenesis.
