基于网络药理学及体外实验探讨桔梗皂苷D对肝癌细胞迁移侵袭的影响及相关机制

Effects of Platycodin D on Migration and Invasion of Hepatocellular Carcinoma Cells Based on Network Pharmacology and In Vitro Experiments

目的基于网络药理学及体外实验方法探讨桔梗皂苷D(PD)对肝癌细胞迁移侵袭的影响及其潜在分子机制。方法检索HERB、PharmMapper及GeneCards数据库预测药物与疾病相关靶标;Venn图构建药物与疾病共同作用靶标;Cytoscape软件建立疾病-药物-靶标相关网络;STRING数据库构建蛋白相互作用网络;Rstuio软件对药物-疾病共同靶标进行基因本体(GO)分析以及京都基因与基因组百科全书(KEGG)信号通路富集分析;CCK-8法检测PD对细胞增殖的作用并筛选出最适给药浓度;伤口愈合实验探究PD对细胞迁移能力的影响;Transwell实验检测PD对细胞迁移及侵袭能力的影响;Western Blot(WB)实验检测相关蛋白表达水平。结果网络药理学显示,PD与肝癌转移过程共有43个共同靶标;GO分析显示,共同靶标的相关途径涉及到调节炎症反应、细胞焦点粘附、细胞-底物连接等关键环节;KEGG分析显示,涉及到的相关通路包括白介素-17信号通路、T细胞受体信号通路等。体外实验显示,PD能够有效抑制肝癌细胞的增殖、迁移及侵袭行为;WB实验显示,PD能够有效上调细胞粘附的关键标志物E-cadherin蛋白的表达。结论PD可能是通过上调E-cadherin蛋白表达,正性调控细胞粘附的生物学过程,从而抑制肝癌细胞的迁移侵袭能力。

Objective To investigate the effect of platycodin D(PD)on the migration and invasion of hepatocellular carcinoma cells and its potential molecular mechanism based on network pharmacology and in vitro experimental methods.Methods Searched HERB,PharmMapper and GeneCards databases to predict drug-related targets;Venn diagram to build drug and disease targets;cytoscape software to establish disease-drug-target network;STRING database to construct protein interaction network;Rstuio software was used for gene ontology(GO)analysis of drug-disease common targets and enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway;CCK-8 assay was used to detect the effect of PD on cell proliferation and the optimal concentration was selected.Wound healing experiment explored the effect of PD on cell migration ability;transwell assay was used to detect the effect of PD on cell migration and invasion;Western Blot(WB)experiment was used to detect the expression levels of related proteins.Results Network pharmacology showed that there were 43 common targets for PD and liver cancer metastasis.GO analysis showed that the related pathways of common targets involved key links such as regulating inflammatory response,cell focus adhesion,and cell-substrate connection.KEGG analysis showed that the involved pathways included interleukin-17 signaling pathway,T cell receptor signaling pathway,etc.In vitro experiments showed that PD could effectively inhibit the proliferation,migration and invasion of HCC cells.WB experiments showed that PD could effectively up-regulate the expression of E-cadherin protein,a key marker of cell adhesion.Conclusion PD may positively regulate the biological process of cell adhesion by upregulating E-cadherin protein expression,thereby inhibiting the migration and invasion ability of hepatocellular carcinoma cells.