Echinacoside Inhibits Breast Cancer Cells By Suppressing Wnt/β-catenin Signaling Pathway

Objective This study is aim to determine the anti-breast cancer ability and the mechanism of Echinacoside by in vitro and in vivo experiments.Methods Firstly,the anti-breast cancer ability of Echinacoside was comprehensively determined by MTT,clony formation,scratch,invasion and migration assays.Then,the luciferase reporter gene system、western blotting and real-time PCR were taken to demonstrate the inhibitory effect of Echinacoside on Wnt/β-catenin signaling pathway.Furthermore,we evaluated the anti-tumor activity of Echinacoside by using an MDA-MB-231 xenograft tumor model,and verified its inhibitory effect on Wnt/β-catenin signaling pathway.Results By using colony formation,scratch,and transwell assays in MDA-MB-231 breast cancer cells,we confirmed the anti-breast cancer ability of Echinacoside in vitro.Then,we found Echinacoside could inhibit reporter gene activity on Wnt1 or LRP6 activate TopFlash system but not on NFATc activate NFAT system.In addition,we found that Echinacoside can dose-dependently reduce phosho-LRP6,total LRP6,phosho-Dvl2,active β-catenin,and total β-catenin protein expression level inMDA-MB-231 and MDA-MB-468 cells by western blot.We also detected well-known Wnt targets genes,including LEF1,CD44,and cyclin D1 by real-time PCR and western blotting,and Echinacoside significantly shows inhibition effect in these two breast cancer cell lines.Furthermore,we investigated its anti-breast cancer ability in an MDA-MB-231 xenograft model in vivo.Echinacoside treatment significantly reduced tumor growth,which was accompanied by a reduction in Wnt/β-catenin signaling.Conclusions In summary,our results demonstrate that Echinacoside can effectively inhibit Wnt/β-catenin signaling,and therefore,it may be a promising therapeutic target to treat breast cancer.