Clone and Expression of VEGF Gene in Human Umbilical Vein Endothelial Cell

Purpose To investigate whether the recombined plasmids that encoding vascular endothelial growth factor 165 (VEGF 165 ) cDNA in an sense or antisense orientation could regulate the secretion level of VEGF protein and affect the growth rate of the transfected human umbilical vein endothelial cell (HUVEC). Methods VEGF 165 cDNA was amplified by reversetranscription PCR (RT?PCR) from fetal human kidney tissue,and was then inserted into the expression plasmid pCR3.1 to construct the recombined plasmids that encoding VEGF 165 cDNA in an sense or antisense orientation.The HUVEC were transfected with these plasmids mediated by liposome.We selected the stable cell lines and observed the morphological characteristics of them under the reversed phase contrast microscope and transmission electron microscope.Growth rates of these cells were acquired by MTT method. Results The recombined plasmids were constructed.The transfected cells were represented just as their parental cells except for the differences in some organelles associated with the protein secretion function.Antisense VEGF decreased the level of secreted VEGF protein in the transtected HUVEC .The inhibitory rate was about 67.7 % ( P 0.01).While the sense VEGF upregulated the secretion level of the transfected cells to 984.8% ( P 0.01 ) than the controls.The growth rates of the transfected HUVEC were changed with the level of its sec retion VEGF protein. Conclusions The recombined plasmids we constructed could successfully regulate the sec retion level of VEGF protein and the growth rate of the transfected HUVEC.