基于网络药理学探讨山奈酚-7-O-新橘皮糖苷抗前列腺癌的作用机制

A network pharmacology-based approach to explore mechanism ofkaempferol-7-O-neohesperidoside against prostate cancer

目的 探讨山奈酚-7-O-新橘皮糖苷(kaempferol-7-O-neohesperidoside, K7ON)抗前列腺癌细胞(prostate cancer, PCa)的作用及潜在分子机制。方法 采用CCK-8法检测K7ON对PCa细胞PC3、DU145、C4-2和LNCap增殖的影响;应用细胞划痕实验检测K7ON对DU145细胞迁移能力的影响;SuperPred等数据库获取K7ON和PCa的靶点;从Venny在线平台获取K7ON与PCa的共同靶点,应用String和Cytoscape构建蛋白相互作用(protein-protein interaction, PPI)网络;通过DAVID数据库进行GO和KEGG功能富集分析,构建“药物-靶点-疾病-通路”网络模型。通过流式细胞术检测K7ON对PCa细胞周期的影响;采用Western blot法检测周期相关蛋白Skp2、p27和p21蛋白的表达;应用Sybyl X2.0将Skp2与K7ON进行分子对接。结果 K7ON可显著抑制PCa细胞的增殖和迁移能力。筛选出药物与疾病的交集靶点共34个,其中Skp2、p27等是K7ON治疗PCa的关键靶点,进一步的GO和KEGG功能功能富集表明其机制主要与细胞周期相关。流式细胞术结果表明,K7ON处理可使DU145细胞周期阻滞在S期。与对照组相比,Skp2蛋白表达水平明显下调,p27和p21的蛋白表达水平上调。分子对接结果显示K7ON与受体Skp2具有较好的结合能力。结论 K7ON可抑制PCa细胞的增殖和迁移,使细胞周期阻滞在S期,其机制可能与调控Skp2-p27/p21信号通路相关。

Aim To explore the effect of kaempferol-7-O-neohesperidoside(K7ON)against prostate cancer(PCa)and the underlying mechanism.Methods The effect of K7ON on the proliferation of PCa cell lines PC3,DU145,C4-2 and LNCaP was detected using CCK8 assay.The effect of K7ON on migration ability of DU145 cells was determined by wound healing assay.The targets of K7ON and PCa were screened from SuperPred and other databases.The common targets both related to K7ON and PCa were obtained from the Venny online platform;a protein-protein interaction network(PPI)was constructed by the String and Cytoscape.Meanwhile,the GO and KEGG functional enrichment were analyzed by David database.Then,a“drug-target-disease-pathway”network model was constructed.Cell cycle of PCa cells treated with K7ON was analyzed by flow cytometry.The expressions of cycle-associated proteins including Skp2,p27 and p21 protein were detected by Western blot.Molecular docking between Skp2 and K7ON was conducted by Sybyl X2.0.Results K7ON significantly inhibited the proliferation and migration of PCa cells.A total number of 34 drug-disease intersection targets were screened.The String results showed that Skp2 and p27,among the common targets,were the key targets of K7ON for PCa treatment.Furthermore,GO and KEGG functional enrichment indicated that the mechanism was mainly related to the cell cycle.Flow cytometry showed that K7ON treatment induced cell cycle arrest at the S phase.Compared with the control group,the protein expression level of Skp2 was significantly down-regulated,while the protein expression levels of p27 and p21 were up-regulated.The network molecular docking indicated that the ligand K7ON had a good binding ability with the receptor Skp2.Conclusions K7ON could inhibit the proliferation and migration of PCa cells,block the cell cycle in the S phase,which may be related to the regulation of cell cycle through the Skp2-p27/p21 signaling pathway.