Inhibitory Effect of Cryptotanshinone on Angiogenesis and Wnt/β-Catenin Signaling Pathway in Human Umbilical Vein Endothelial Cells

• Published in: Chinese journal of integrative medicine Vol. 20; no. 10; pp. 743 - 750
• Main Author: 陈倩 庄钦 毛威 徐晓明 王丽慧 王海兵
• Format: Journal Article
• Language: English
• Published: Heidelberg Chinese Association of Traditional and Western Medicine 01.10.2014
• Subjects: beta Catenin - metabolism; Blotting, Western; Cell Movement - drug effects; Cell Survival - drug effects; Cyclin D1 - metabolism; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Immunohistochemistry; Luciferases - metabolism; Medicine; Medicine & Public Health; Neovascularization, Physiologic - drug effects; Original Article; Phenanthrenes - chemistry; Phenanthrenes - pharmacology; Vascular Endothelial Growth Factor A - metabolism; Western印迹; Wnt; Wnt Signaling Pathway - drug effects; 人脐静脉内皮细胞; 信号通路; 抑制作用; 抗血管生成; 细胞周期蛋白D1; 隐丹参酮; atherosclerosis; cryptotanshinone; Wnt/β-catenin signaling pathway; human umbilical vein endothelial cell; angiogenesis
• ISSN: 1672-0415; 1993-0402
• DOI: 10.1007/s11655-014-1810-x

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.