Mechanism of treating gastric cancer with Shenzhu Baihu Granules based upon a reversal of methylation modification
OBJECTIVE To predict and verify the methylation regulatory sites of Shenzhu Baihu Granules(Shenzhu)and explore its mechanism of action in treating gastric cancer(GC).METHODS Anti-tumor effect of Shenshu was verified by a NOD scid gamma(NSG)nude murine model of GC.The effective chemical components and surgical targets were mined by searching the databases of Traditional Chinese Medicine Systems Pharmacology Database & Analysis Platform(TCMSP),Drug-Bank,UniProt and PubChem and the differentially expressed genes of GC retrieved from the databases of GeneCards,MalaCards and GEO.The potential surgical targets for GC were obtained by intersection of both with Venny 2.1.0.A protein interaction network diagram was constructed in the database of STRING 11.5.GO functional and KEGG pathway enrichment analyses were performed with the database of DAVID.And a"drug component target disease pathway"network diagram was constructed with Cytoscape software for screening out key pathways and core targets for treating GC.Molecular docking verification of active ingre-dients and core targets was performed by AutoDockTools 1.1.2 software.And visualization of mapping results was enabled by Pymol software.Finally real-time quantitative polymerase cahin reaction(RT-qPCR),Western blot and immunohistochemistry were utilized for verifying target genes.RESULTS After 12-day dosing,tumor volume and weight of treatment group were sig-nificantly smaller than those of control group(P＜0.05).A total of 42 effective drug ingredients were screened through network pharmacology with 664 potential target genes.After intersecting with 71 differentially expressed genes in GC,15 target genes were obtained.GO/KEGG analysis revealed 11 pathways and two major genes for methylation modification-DNMT1 and EZH2.There was a significant marked down-regulation of DNMT1/EZH2 in tumor tissue of treatment group(P＜0.05).Molecular docking revealed that 7 active drug ingredients possessed docking activity with DNMT1/EZH2.CONCLUSION Seven key active ingredients in Shenshu,including acacia,luteolin and quercetin,may act on DNMT1/EZH2 core target proteins,reverse DNA/histone methylation modification and exert inhibitory effects on GC growth.
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