Reconstruction of Human Vascular Endothelial Cells with Antisense Nucleic Acid Technology

In early stages of the formation of thrombus,the von Willebrand factor(vWF) secreted by vascular endothelial cells-connects the platelet and the wounded site of inner wall of the vascellum and then originates the cascade reaction of blood coagulation.Vascular endothelial cells in which the expression of vWF is inhibited effectively can be constructed using antisense nucleic acid technology.An antisense nucleic acid fragment of vWF was artificially synthesized and then subcloned into eukaryotic expression plasmid pcDNA3.1(+) in the reverse direction.The plasmid pcDNA3.1(+)/pvWF was transfected by lipofectamine into vascular endothelial cells after being sequenced to confirm the correction of gene.Transgenic cells were cultured in large scale after the selection using neomycin solution.Method of RT-PCR was used to investigate the vWF mRNA expression of transgenic cells.The result showed that the eukaryotic expression plasmid of vWF antisense nucleic acid could inhibit the vWF mRNA expression effectively.So a vascular endothelial cells model was set up that can express vWF antisense nucleic acid and the work provided a genetic reforming cell for the research of anticoagulated blood materials.