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).
Conclusion
Our study supported the role of CPT as an angiogenic inhibitor, which may impact on the Wnt/β-catenin signaling pathway.
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Supported by National Natural Science Foundation of China (No. 81170270) and Medicine and Technology Program of Zhejiang Province (No. 2013KYB188)
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Chen, Q., Zhuang, Q., Mao, W. et al. Inhibitory effect of cryptotanshinone on angiogenesis and Wnt/β-catenin signaling pathway in human umbilical vein endothelial cells. Chin. J. Integr. Med. 20, 743–750 (2014). https://doi.org/10.1007/s11655-014-1810-x
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DOI: https://doi.org/10.1007/s11655-014-1810-x