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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 μmol/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 μmol/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 p-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 LiCI(an activator of the pathway) treatment.However,the signals were weaker in the nucleus of the CPT(10 μmol/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).Conclosion:Our study supported the role of CPT as an angiogenic inhibitor,which may impact on the Wnt/β-catenin signaling pathway.
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 μmol / 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 μmol / L CPT for detecting endothelial cell migration, invasion, and tubular-like structure formation with wound healing, transwell invasion and matrigel tube formation assays, respectively. 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 p-catenin was evaluated using Western blot and immunochemistry. Finaly, vascular endothelial growth factor (VEGF) and cyclin D1, downstream proteins of the Wnt pathway were examined with Western blot.Resu lts: 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 LiCI (an activator of the pathway) treatment.However, the signals were weaker in the nucleus of the CPT (10 μmol / L) group, compared to the positive control. Als, VEGF and cyclin D1 were both eliminated by CPT in 5 and 10 μ mol / L doses (P <0.05) .Conclosion: Our study supported the role of CPT as an angiogenic inhibitor, which may impact on the Wnt / β-catenin signaling pathway.