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


CONCLUSIONS
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.
Department of Neurosurgery, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan, ROC.
Li
L
Wei
Division of Nephrology, Department of Internal Medicine, Tri-Service General Hospital, Taipei, Taiwan, ROC.
Jin-Shuen
JS
Chen
Department of Ophthalmology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, ROC.
Hsin-Ting
HT
Lin
Division of Endocrinology and Metabolism, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, ROC.
Chung-Ze
CZ
Wu
Division of Nephrology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, ROC.
Cai-Mei
CM
Zheng
School of Pharmacy and Graduate Institute Pharmacy, National Defense Medical Center, Taipei, Taiwan.
Yu-Ching
YC
Chang
School of Pharmacy and Graduate Institute Pharmacy, National Defense Medical Center, Taipei, Taiwan.
Li-Chien
LC
Chang
Division of Nephrology, Department of Internal Medicine, Tri-Service General Hospital, Taipei, Taiwan, ROC; Division of Nephrology, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, ROC; Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan, ROC. Electronic address: linyf@s.tmu.edu.tw.
Yuh-Feng
YF
Lin
D000199:Actins / Q000032:analysis / Q000502:physiology*; D000818:Animals; D000069059:Atorvastatin / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D051379:Mice; D010957:Plasmids; D055293:Receptors, Urokinase Plasminogen Activator / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects*
D000199:Actins; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D055293:Receptors, Urokinase Plasminogen Activator; D000069059:Atorvastatin
angiogenesis; atorvastatin; cytoskeleton; urokinase receptor