Anti-liver Cancer Mechanism of Hypericin Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo explore the mechanism of hypericin against liver cancer using network pharmacology. MethodThe traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP）， Drug Gene Interaction Database （DGIdb）， Comparative Toxicogenomics Database （CTD） and SwissTargetPrediction were used to predict the targets of hypericin. Five databases including GeneCards and Online Mendelian Inheritance in Man （OMIM） were employed to obtain liver cancer-related targets. The intersection was performed to obtain the targets of hypericin against liver cancer. The Database for Annotation， Visualization and Integrated Discovery （DAVID） v2021q4 was used for Gene Ontology （GO） function annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. The protein-protein interaction （PPI） network of the targets was constructed by Cytoscape 3.6.1 to screen the core targets，and the affinity between hypericin and the core targets was verified by molecular docking. The effects of hypericin on liver cancer and the migration of liver cancer cells were observed by cell viability assay and would healing assay， respectively， and its effects on the mRNA and protein expression of key targets cysteinyl aspartate-specific protease-3（Caspase-3） and mitogen-activated protein kinase 3 （MAPK3） were detected by real-time polymerase chain reaction（Real-time PCR） and Western blot， respectively. ResultA total of 45 genes related to the anti-liver cancer effect of hypericin were obtained， and six core target genes were screened. The signal pathways enriched by KEGG pathway analysis included apoptosis，tumor necrosis factor （TNF） and cancer signal pathways. Molecular docking showed that the core target genes Caspase-3，TNF，estrogen receptor 1 （ESR1），MAPK3，catalase （CAT） and cyclooxygenase 2 （PTGS2） had good affinity with hypericin，especially Caspase-3 and MAPK3. In addition，compared with the conditions in control group， cell experiments demonstrated that hypericin could reduce the viability of liver cancer cells （P<0.05），inhibit their migration，increase the mRNA expression of Caspase-3 （P<0.05） and decrease that of MAPK3 （P<0.05）. ConclusionHypericin exerted the anti-liver cancer effect by affecting the core targets such as Caspase-3，TNF，ESR1，MAPK3，CAT and PTGS2 and jointly interfering with apoptosis，TNF and cancer signal pathways.