title: Coral-derived compound WA-25 inhibits angiogenesis by attenuating the VEGF/VEGFR2 signaling pathway.
abstract: BACKGROUND
  WA-25 (dihydroaustrasulfone alcohol, a synthetic derivative of marine compound WE-2) suppresses atherosclerosis in rats by reducing neointima formation. Because angiogenesis plays a critical role in the pathogenesis of atherosclerosis, the present study investigated the angiogenic function and mechanism of WA-25.
  
  
  METHODS
  The angiogenic effect of WA-25 was evaluated using a rat aortic ring assay and transgenic zebrafish models were established using transgenic Tg(fli-1:EGFP)y1 and Tg(kdrl:mCherryci5-fli1a:negfpy7) zebrafish embryos. In addition, the effect of WA-25 on distinct angiogenic processes, including matrix metalloproteinase (MMP) expression, endothelial cell proliferation and migration, as well as tube formation, was studied using human umbilical vein endothelial cells (HUVECs). The effect of WA-25 on the endothelial vascular endothelial growth factor (VEGF) signaling pathway was elucidated using qRT-PCR, immunoblot analysis, immunofluorescence and flow cytometric analyses.
  
  
  RESULTS
  The application of WA-25 perturbed the development of intersegmental vessels in transgenic zebrafish. Moreover, WA-25 potently suppressed microvessel sprouting in organotypic rat aortic rings. Among cultured endothelial cells, WA-25 significantly and dose-dependently inhibited MMP-2/MMP-9 expression, proliferation, migration and tube formation in HUVECs. Mechanistic studies revealed that WA-25 significantly reduced the VEGF release by reducing VEGF expression at the mRNA and protein levels. In addition, WA-25 reduced surface VEGF receptor 2 (VEGFR2/Flk-1) expression by repressing the VEGFR2 mRNA level. Finally, an exogenous VEGF supply partially rescued the WA-25-induced angiogenesis blockage in vitro and in vivo.
  
  
  CONCLUSIONS
  WA-25 is a potent angiogenesis inhibitor that acts through the down-regulation of VEGF and VEGFR2 in endothelial cells.
  
  
  CONCLUSIONS
  WA-25 may constitute a novel anti-angiogenic drug that acts by targeting endothelial VEGF/VEGFR2 signaling.
authors: Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University and Academia Sinica, Kaohsiung 804, Taiwan. shiwey@yahoo.com.tw.
authors: Shih-Wei
authors: SW
authors: Lin
authors: Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan. sghung@hotmail.com.|Department of Internal Medicine, Kaohsiung Armed Forces General Hospital, Kaohsiung 802, Taiwan. sghung@hotmail.com.
authors: Shih-Chung
authors: SC
authors: Huang
authors: Mitochondrial Research Unit, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 833, Taiwan. hsiaomeikuo@gmail.com.
authors: Hsiao-Mei
authors: HM
authors: Kuo
authors: Division of Nephrology Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 833, Taiwan. becky0917@yahoo.com.tw.
authors: Chiu-Hua
authors: CH
authors: Chen
authors: Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. ylma@vghks.gov.tw.|Division of Nephrology, Kaohsiung Veterans General Hospital, Kaohsiung 813, Taiwan. ylma@vghks.gov.tw.
authors: Yi-Ling
authors: YL
authors: Ma
authors: Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan. skbboyz0817@gmail.com.
authors: Tian-Huei
authors: TH
authors: Chu
authors: Department of Ophthalmology, Kaohsiung Veterans General Hospital, Kaohsiung 804, Taiwan. ysbee@vghks.gov.tw.
authors: Youn-Shen
authors: YS
authors: Bee
authors: Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. emwang@vghks.gov.tw.
authors: E-Ming
authors: EM
authors: Wang
authors: Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. cywu@mail.nsysu.edu.tw.
authors: Chang-Yi
authors: CY
authors: Wu
authors: National Museum of Marine Biology and Aquarium, Pingtung 944, Taiwan. pjsung@nmmba.gov.tw.
authors: Ping-Jyun
authors: PJ
authors: Sung
authors: Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University and Academia Sinica, Kaohsiung 804, Taiwan. wzh@mail.nsysu.edu.tw.|Department of Marine Biotechnology and Resources National Sun Yat-Sen University, Kaohsiung 804, Taiwan. wzh@mail.nsysu.edu.tw.
authors: Zhi-Hong
authors: ZH
authors: Wen
authors: Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 807, Taiwan. dechwu@yahoo.com.|Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. dechwu@yahoo.com.|Department of Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan. dechwu@yahoo.com.
authors: Deng-Chyang
authors: DC
authors: Wu
authors: Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University and Academia Sinica, Kaohsiung 804, Taiwan. sheu@mail.nsysu.edu.tw.|Department of Marine Biotechnology and Resources National Sun Yat-Sen University, Kaohsiung 804, Taiwan. sheu@mail.nsysu.edu.tw.|Department of Medical Research, China Medical University Hospital, China Medical University, Taichung 404, Taiwan. sheu@mail.nsysu.edu.tw.
authors: Jyh-Horng
authors: JH
authors: Sheu
authors: Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University and Academia Sinica, Kaohsiung 804, Taiwan. minghongtai@gmail.com.|Institute of Biomedical Sciences, National Sun Yat-sen University, Kaohsiung 804, Taiwan. minghongtai@gmail.com.|Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung 807, Taiwan. minghongtai@gmail.com.
authors: Ming-Hong
authors: MH
authors: Tai
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D037421:Anthozoa / Q000737:chemistry*; D002074:Butanones / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D006801:Humans; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D009389:Neovascularization, Pathologic / Q000517:prevention & control; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D013450:Sulfones / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000187:drug effects*; D015027:Zebrafish
chemical_list: D020533:Angiogenesis Inhibitors; D002074:Butanones; D061965:Matrix Metalloproteinase Inhibitors; D013450:Sulfones; D042461:Vascular Endothelial Growth Factor A; C000590752:dihydroaustrasulfone alcohol; D040301:Vascular Endothelial Growth Factor Receptor-2