Effect of adenovirus‑mediated uPA gene transduction on the fibrinolytic activity of human umbilical vein endothelial cells.

Abstract

The present study aimed to investigate the effect of adenovirus‑mediated urokinase‑type plasminogen activator (uPA) transduction on uPA expression and fibrinolytic activity in human umbilical vein endothelial cells (HUVECs). Recombinant adenovirus vectors containing the human uPA gene were constructed and transduced into HUVECs. The expression and fibrinolytic activity of uPA was then assessed in HUVECs using western blot analysis, ELISA and colorimetric assay. The experiments were performed in three groups: The ad/uPa (n=3), ad/neg control (n=3) and blank control (n=3) groups. Western blot analysis revealed that uPA protein expression in the HUVECs in the ad/uPa group was significantly increased compared with those in the ad/neg control or blank groups (P<0.01). The uPA protein levels in the supernatant of the three groups were 379.40±2.46, 240.01±1.16 and 256.10±3.04ng/l, respectively, showing that the uPA protein levels were significantly higher in the supernatant in the ad/uPa group compared with those in the ad/neg control or blank groups. uPA activity was determined using a colorimetric method and was found to be 40238.49±5755IU/mg in the HUVECs in the ad/uPa group, which was significantly higher than that in the HUVECs in the ad/neg control (6180.03±942.38IU/mg) or blank groups (3346.06±928.81IU/mg) (both P<0.01). These findings suggested that transduction of the uPA gene increased uPA protein expression and fibrinolytic activity in HUVECs.