Screening Of Wnt Inhibitors And Evaluation Of The Anti-gastric Cancer Effect

Objective: The Wnt/β-catenin signaling pathway has an integral role in embryonic development,cell cycle,inflammatory response and tumor formation.More than 50%of gastric cancer(GC)cases have dysregulated Wnt/β-catenin signaling pathway,and about 30% of gastric cancer patients have β-catenin nuclear aggregation.However,the pharmacological efficacy,toxicity properties,and molecular mechanisms of agents targeting the Wnt pathway in GC remain ambiguous.This study attempted to screen the optimal Wnt inhibitors and explore its properties in terms of toxicology and effects on GC biological behavior.Methods: The efficacy of Wnt inhibitors was evaluated in three poorly differentiated GC cell lines with aberrant activation of the Wnt pathway.Colony formation,wound healing assay,transwell assay,and flow cytometry were performed to characterize the effects of these inhibitors on the metastasis,invasion,and apoptosis of GC cells,respectively.The toxicity profiles of the inhibitors were assessed using the admet SAR server.The expression of EMT-related proteins was determined using Western blot and immunohistochemical staining.Furthermore,cell-derived xenograft(CDX)models were used to evaluate the potencies of the identified inhibitor in gastric cancer progression.Results: The expression of β-catenin,the key protein of the Wnt pathway,is significantly upregulated in GC and it negatively correlates with the prognosis of poorly differentiated GC.Through screening 12 inhibitors of the Wnt pathway,we discovered that NCB-0846,Niclosamide,and PRI-724 showed significant potency to suppress GC cells growth.Among all the screened molecules,NCB-0846 exhibited the optimal potency against GC.It blocked the GC cell invasion and migration,and promoted the apoptosis of GC cells in vitro.In particular,the prediction of ADMET(Absorption,Distribution,Metabolism,Excretion,and Toxicity)properties for NCB-0846 did not reflect the risk of mutagenesis and carcinogenesis.Importantly,NCB-0846 exhibited a potent anti-tumor efficacy in CDX models and negatively regulated epithelialmesenchymal transformation(EMT).Conclusions: These results illustrate that NCB-0846 had significant anti-gastric cancer effect and reduced the invasion and migration ability of gastric cancer by inhibiting epithelial-mesenchymal transformation.