Atorvastatin from target screening attenuates endothelial cell tube formation and migration by regulating urokinase receptor-related signaling pathway and F/G actin

Abstract

Background: Angiogenesis and cytoskeletal transformation are common denominators of many pathological developments. The relationship between angiogenesis, urokinase plasminogen activator receptor (uPAR) signaling pathway, and cytoskeletal transformation is still unknown. In this study, a pGL3-uPAR promoter reporter system combined with Bio-Plex mRNA analysis was established for discovering uPAR modulators to analyze this interconnection.

Methods: After screening a set of clinically used drugs, atorvastatin (Ator) was found to significantly affect uPAR expression and its ideal dose, 1 μM, was determined for cell study. Mouse endothelial cell (mEC) models, including tube formation for angiogenesis and wound healing assay for migration, were employed to test the effects on angiogenesis and cytoskeletal transformation with (Group Ator) and without (Group C) the treatment of Ator.

Results: The mEC tube formation and migration was significantly decreased in Group Ator. Regarding cytoskeleton changes, the ratio of F/G actin by Western blotting and the assembly of F-actin (lamellipodia) by immunofluorescence were attenuated. Furthermore, uPAR and all uPAR-related factors, including integrin α5β3, phosphorylated-focal adhesion kinase, and Rac, revealed a significant reduction when compared with Group C.

Conclusion: We conclude that close regulatory machinery spans angiogenesis, uPAR signaling, and cytoskeletal transformation, and that uPAR modulator Ator can decrease the reorganization of actin cytoskeleton, which may lead to a new approach in angiogenesis.

Keywords: angiogenesis; atorvastatin; cytoskeleton; urokinase receptor.