基于网络药理学和细胞验证探讨美迪紫檀素通过p53依赖性周期调控通路抑制肝癌细胞增殖的作用机制OA北大核心CSTPCD

Objective To investigate the proliferation inhibition effect induced by medicarpin in liver cancer cells and its mechanism.Methods HepG2,HCC-M,and Hep3B cell lines were used to investigate the impact of medicarpin on the proliferation and survival of liver cancer cells.Changes(50 μmol/L medicarpin)in cell morphology were observed by inverted microscope and the proliferative capacity of cells(12.5,25,50 μmol/L medicarpin)was assessed using colony formation assays.To explore the potential mechanisms of medicarpin,the Swiss Target Prediction database was used to predict its target proteins.Subsequently,protein-protein interaction networks were constructed using STRING and Cytoscape,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Flow cytometry assay was used to evaluate cell cycle progression,while Western blotting was performed to assess the expression levels of cell cycle-related proteins.Furthermore,the effects of medicarpin on different cell lines with various levels of p53 expression were compared to validate the role of p53 in medicarpin-induced cell cycle arrest in liver cancer cells.Results Morphological changes such as disrupted membrane integrity,nuclear shrinkage,and loss of intercellular connections were observed in liver cancer cells treated with medicarpin.Additionally,a significant decrease in cell numbers was evident at various concentrations(12.5,25,50 μmol/L),and colony formation assays indicated that medicarpin significantly inhibited the quantity of colonies formed by liver cancer cells(P<0.05).The network construction and functional enrichment analysis indicated that medicarpin targets are enriched in cell cycle regulatory signaling pathways.Flow cytometry result showed that medicarpin induced G2/M cell cycle arrest in HepG2 cells,accompanied by the accumulation of p53 protein and downregulation of the cell cycle regulator cdc2(P<0.05).However,the G2/M phase cell cycle arrest induced by medicarpin could be reversed by a p53 inhibitor.More importantly,medicarpin was unable to induce G2/M phase cell cycle arrest in Hep3B(p53-null)and p53 knockout HCT116 cells,consistently indicating the critical role of p53 in medicarpin-induced G2/M phase cell cycle arrest.Conclusion Medicarpin inhibits the proliferation of liver cancer cells,and its anticancer effect is related to p53-dependent G2/M cell cycle arrest.