Mechanism of tetrandrine in treatment of pancreatic adenocarcinoma based on network pharmacology,molecular docking and in vitro experiments
Objective To analyze the potential therapeutic targets of tetrandrine(TET)in treatment of pancreatic adenocarcinoma(PAAD)by network pharmacology,molecular docking,and in vitro cellular test,and to clarify the related molecular mechanism.Methods Potential target datasets for TET were compiled utilizing the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP),Swiss Target Prediction and PharmMapper.To obtain target information pertinent to pancreatic cancer,the term"pancreatic ductal adenocarcinoma"was employed in searches within the GeneCards and OMIM databases.The intersection of TET targets and pancreatic cancer targets was identified to determine the potential therapeutic targets for pancreatic cancer.Subsequently,a protein-protein interaction(PPI)network was constructed using the STRING database and imported into Cytoscape 3.8.0 software to identify core targets.Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were conducted using the Metascape database.Molecular docking validation was performed utilizing AutoDock and PyMOL software.Cell proliferation activity was detected by CCK-8 assay.Additionally,pancreatic cancer cells were categorized into a control group and TET treatment group(6.25,12.5,25 μmol/L)to evaluate cell migration via the scratch assay,clone formation ability through colony formation assay,and cell cycle distribution using flow cytometry.The mRNA expression on level of PI3K-Akt pathway-related genes in the cells were detected using the real-time fluorescence quantitative PCR(qRT-PCR)method.Results A total of 140 potential targets of TET affecting pancreatic cancer were obtained,the PPI network between TET and pancreatic cancer was analyzed and constructed,and core targets such as Akt1,epidermal growth factor receptor(EGFR)and PIK3CA were obtained.Molecular docking showed that these core targets had good binding activity with TET.GO and KEGG enrichment analysis revealed that the key targets of TET in the treatment of pancreatic cancer were mainly involved in biological processes such as phosphorylation and PI3K-Akt,MAPK and other signaling pathways.It was observed that the mortality rate of pancreatic cancer cells treated with TET increased proportionally with the concentration of the drug,compared to the control group.Additionally,the migration rate of PANC-1 cells in the TET treatment group was significantly reduced(P<0.01)relative to the control group.Furthermore,compared with the blank control group,the number of colony formation of PANC-1 cells in the TET treatment group was significantly decreased(P<0.001),indicating a dose-dependent effect.Flow cytometry analysis revealed a significant increase in the proportion of cells in the G0/G1 phase in the TET treatment group compared with control group(P<0.001).Compared with the control group,the expression levels of PDGFRA mRNA in the TET group were significantly decreased(P<0.05).The mRNA expression levels of TCL1A,Jun,ILK,mTOR,NF-κBIA,GRB10,FOS,CASP9,PIK3CA,CD40,and MAPK8 were significantly increased(P<0.05).Conclusion TET can inhibit PAAC cell proliferation,migration,cloning,and causing cell cycle arrest,and the underlying mechanism of which may involve the PI3K-Akt signaling pathways.
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