title: Inhibitory effect of cryptotanshinone on angiogenesis and Wnt/β-catenin signaling pathway in human umbilical vein endothelial cells.
abstract: OBJECTIVE
  To investigate the anti-angiogenic effect of cryptotanshinone (CPT) on human umbilical vein endothelial cells (HUVECs) and the effect of CPT on Wnt/β-catenin signaling pathway.
  
  
  METHODS
  HUVECs were incubated with 0, 2.5, 5, 10, and 20 μ mol/L CPT for detecting cell viability with dimethyl thiazolyl-2,5-diphenyltetrazolium bromide (MTT) assay. Then, HUVECs were incubated with 0, 2.5, 5, and 10 μ mol/L CPT for detecting endothelial cell migration, invasion, and tubular-like structure formation with wound healing, transwell invasion and matrigel tube formation assays, respectively. To gain insight into CPT-mediated signaling, the effects of CPT on T-cell factor/lymphocyte enhancer factor (TCF/LEF) transcription factors were detected by the Dual-luciferase reporter assay. Next, the nuclear expression of β-catenin was evaluated using Western blot and immunochemistry. Finally, vascular endothelial growth factor (VEGF) and cyclin D1, downstream proteins of the Wnt pathway were examined with Western blot.
  
  
  RESULTS
  CPT dose-dependently suppressed endothelial cell viability, migration, invasion, and tubular-like structure formation. In particular, CPT blocked β-catenindependent transcription in HUVECs in a dose-dependent manner. In Western bolt, 10 μ mol/L CPT decreased expression of β-catenin in nucleus of HUVECs (P<0.01). In immunohistochemistry, β-catenin was more potent in response to LiCl (an activator of the pathway) treatment. However, the signals were weaker in the nucleus of the CPT (10 μ mol/L) group, compared to the positive control. Also, VEGF and cyclin D1 were both eliminated by CPT in 5 and 10 μ mol/L doses (P<0.05).
  
  
  CONCLUSIONS
  Our study supported the role of CPT as an angiogenic inhibitor, which may impact on the Wnt/β-catenin signaling pathway.
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, 310006, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhuang
authors.forename: Wei
authors.initials: W
authors.lastname: Mao
authors.forename: Xiao-ming
authors.initials: XM
authors.lastname: Xu
authors.forename: Li-hui
authors.initials: LH
authors.lastname: Wang
authors.forename: Hai-bing
authors.initials: HB
authors.lastname: Wang
mesh_terms: D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019938:Cyclin D1 / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008156:Luciferases / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010616:Phenanthrenes / Q000737:chemistry / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects*; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D010616:Phenanthrenes; D042461:Vascular Endothelial Growth Factor A; D051176:beta Catenin; D019938:Cyclin D1; C037886:cryptotanshinone; D008156:Luciferases
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