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 24 h and the cytotoxicity was determined by 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, we first studied effects of CPT on TCF/LEF transcription factors 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 cyclinD1, 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-catenin-dependent transcription in HUVECs in a dose-dependent manner.Using Western bolt, we showed here that 10 μmol/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 μmol/L) group, compared to the positive control.Also, VEGF and cyclinD1 were both eliminated by CPT in 5 and 10 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.