Effects of hVEGF_(121 ) gene transfection on human umbilical vein endothelial cells growth

Objective:To construct human vascular endothelial growth factor (hVEGF)-121 protein expression plasmid, and to transfect it into human umbilical vein endothelial cells (HUVEC) to speed up their growth in vitro through a paracrine way. Methods: hVEGF 121 cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR) from human embryonic fibroblast and was inserted into eukaryotic expression vector pcDNA3. The recombinant plasmid was tranfected into HUVEC by FuGENE 6 liposome. The culture supernatant of the transfected cells was collected, and the expression of hVEGF protein was tested by enzyme-linked immunosorbent assay (ELISA). The role of hVEGF 121 gene transfer in HUVEC proliferation was studied by cell-counting. pcDNA3 vector transfected and non-transfected HUVEC were used as the control groups. Results: pcDNA3-hVEGF 121 plasmid was constructed. After cell transfection, hVEGF protein was transiently expressed and peaked on 1 d at 59.95 pg/(10 4 cells·d), about 100 times that of the control group level; afterwards, the productions decreased quickly and then slowly, but still about 1 time higher than that of the control group level on 7 d. The transfected HUVEC proliferated more rapidly than that of the control groups on 3 d (P0.01),with the cell doubling time shortened to about 3 d from over 7 d in the control group.Conclusion:Plasmid pcDNA3- hVEGF 121 has been constructed successfully, which can express active hVEGF 121 protein in HUVEC cultured in vitro,thus promoting HUVEC proliferation by a paracrine loop.