Based on biological data and experimental research Wen-Yang-Hua-Liu Decoction conducted a mechanistic study on the inhibition of non-small cell lung cancerOA

Background:The mortality rate of non-small cell lung cancer(NSCLC)has continued to rise in recent decades,and the five-year survival rate remains extremely low,despite the accelerated incorporation of a variety of medical treatments in its treatment and the increasing incidence of lung cancer.Traditional Chinese medicine plays an important role in the prevention and treatment of lung cancer.We used network pharmacology to investigate and predict the targets and associated signaling pathways of Wen-Yang-Hua-Liu Decoction(WYHLD)for the treatment of NSCLC.In addition,we investigated the growth mechanism of WYHLD on NSCLC by observing the effects of WYHLD on the processes of NSCLC proliferation,apoptosis,and ferroptosis.Methods:WYHLD drugs were obtained from the TCMSP database and potential targets from the Swiss Target Prediction and Uniprot databases;NSCLC-related genes were collected from the Genecards database;and the intersection genes of WYHLD and NSCLC were obtained from the Venny2.1 platform and imported into the STRING database.The intersection genes of WYHLD and NSCLC were collected from the Genecards database,imported into the STRING database,constructed into a into a protein-protein interaction network(PPI),visualized by Cytoscape 3.9.1 software,and analysed by GO enrichment and KEGG pathway analysis using the Metascape database to predict the direct targets and signaling pathways of WYHLD in the treatment of NSCLC.The NSCLC cell line A549 was cultured,and CCK-8 and wound healing assays were performed to assess cell proliferation and migration.Apoptosis was detected by Hoechst staining.Reactive oxygen species(ROS)levels were measured using flow cytometry.The microstructure of A549 cells observed by transmission electron microscopy.In addition,Western blot was performed to detect MYC,VEGF,HIF-1α,β-catenin,Cleaved-caspase3,GPX4,and SLC7A11 protein expression.Results:The network pharmacological analysis identified 160 WYHLD compounds,1,444 NSCLC disease-related targets,and 133 overlapping WYHLD and NSCLC targets.MYC,AKT1,JUN,ESR1,FOS,TP53,BCL2,and other proteins with high degree values were ranked as therapeutic targets in the STRING 11.0 database.The enriched pathways were identified as being associated with the cancer pathway,the AGE-RAGE signaling pathway,and NSCLC.In in vitro experiments,we found that WYHLD inhibited the migration and proliferation of NSCLC cells and promoted their apoptosis.Analysis of reactive oxygen species showed that WYHLD could promote cellular production of reactive oxygen species.In addition,WYHLD significantly suppressed the expression of MYC,GPX4,SLC7A11,β-catenin,and VEGF while increasing the expression of Cleaved-caspase3.Conclusion:Our results suggest that WYHLD may act as a novel strategy for the prevention and treatment of lung cancer by targeting the Wnt pathway,thereby inhibiting the growth mechanism of NSCLC.