title: Fasudil inhibits prostate cancer-induced angiogenesis in vitro.
abstract: Inhibition of angiogenesis is an important therapeutic strategy for advanced stage prostate cancer (PCa). RhoA/Rho-associated protein kinases (ROCK) are key regulators of the cytoskeleton and have been implicated in PCa angiogenesis. We investigated the anti-angiogenic effects of fasudil, a ROCK inhibitor, on PCa-induced angiogenesis in vitro. Proliferation of PCa-conditioned human umbilical vein endothelial cells (HUVECs) was assessed using a bromodeoxyuridine (BrdU) assay, and migration was assessed with a wound healing assay. In vitro angiogenesis of PCa-conditioned HUVECs was evaluated by tube formation and a spheroid sprouting assay. Fasudil inhibited PCa-induced endothelial cell proliferation at a concentration of 100 µM, and also decreased PCa-induced endothelial cell migration at a concentration of 30 µM. In the in vitro angiogenesis assay, fasudil exerted a more significant effect. Tube formation was significantly inhibited at fasudil concentrations exceeding 3 µM, and spheroid sprouts were significantly thinner and shorter (at fasudil concentrations of 10 and 30 µM, respectively). Western blotting results showed that expression of phosphorylated myosin phosphatase target subunit 1 (MYPT-1) was significantly lower after fasudil treatment, confirming that fasudil inhibited ROCK activity in these model systems. These data suggest that fasudil may be a useful anti-angiogenic agent for PCa.
authors.affiliation: Medical School, Service de Physiologie, Paris Descartes University, EA 2511, Hôpital Cochin, 75014 Paris, France.
authors.forename: Weihua
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Urology, Shanghai East Hospital, Tongji University, School of Medicine, Shanghai 200120, P.R. China.
authors.forename: Kaili
authors.initials: K
authors.lastname: Mao
authors.affiliation: Medical School, Service de Physiologie, Paris Descartes University, EA 2511, Hôpital Cochin, 75014 Paris, France.
authors.forename: Thong
authors.initials: T
authors.lastname: Hua-Huy
authors.affiliation: Medical School, Service de Physiologie, Paris Descartes University, EA 2511, Hôpital Cochin, 75014 Paris, France.
authors.forename: Yihua
authors.initials: Y
authors.lastname: Bei
authors.affiliation: Clinical and Translational Research Center, Shanghai East Hospital, Tongji University, School of Medicine, Shanghai 200120, P.R. China.
authors.forename: Zhongmin
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Medical School, Service de Physiologie, Paris Descartes University, EA 2511, Hôpital Cochin, 75014 Paris, France.
authors.forename: Anh-Tuan
authors.initials: AT
authors.lastname: Dinh-Xuan
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000008:administration & dosage / Q000031:analogs & derivatives*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008297:Male; D043263:Myosin-Light-Chain Phosphatase / Q000096:biosynthesis; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: D047428:Protein Kinase Inhibitors; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D054460:rho-Associated Kinases; D043263:Myosin-Light-Chain Phosphatase; C490372:PPP1R12A protein, human; C049347:fasudil