Objective To investigate the effects of overexpression of von Willebrand factor (vWF) on the proliferation, migration, invasion, apoptosis, and tube formation of human umbilical vein endothelial cells (HUVECs).
Methods Five vWF-overexpressing plasmid CRISPRa vectors were designed and constructed, and HUVECs were divided into the experimental group (5 groups) and control group.Five plasmids were transfected into the experimental group, and the expression effect of vWF in each group was detected by Western blotting, and the dominant group was selected.The effects of vWF overexpression on the proliferation, migration, invasion, apoptosis and tube formation of HUVECs were observed by cell proliferation assay, scratch assay, cell invasion assay, apoptosis assay, and cell tube formation assay.Western blotting was used to detect the expression of 5 common pathway protein factors in the two groups of cells and explore the possible related mechanisms.
Results Western blotting showed that the expression level of vWF in HUVECs after successful transfection was significantly higher than that in the control group (P < 0.05).Among them, the overexpression of vWF in the vWF-OE4 group was significantly higher than that of other groups, so the primer sequence was selected for subsequent experiments.Compared with the control group, the cell proliferation in vWF-OE group was slowed down, the cell scratch 48 h mobility was decreased, the Transwell cell invasion rate was decreased (P < 0.05);the apoptosis rate, angiogenesis ability and the length of HUVECs producing tubules were increased (P < 0.05);the protein contents of JUN and P53 were increased (P < 0.05).
Conclusions Overexpression of vWF can significantly inhibit the proliferation, migration and invasion of HUVECs, promote apoptosis and enhance the ability of tube formation.
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