摘要
目的:探究平消胶囊治疗乳腺癌的潜在作用机制.方法:利用TCMSP、TCM-ID、GeneCards等数据库筛选平消胶囊与乳腺癌的相关靶点;Cyto-scape软件构建"药物-靶点-疾病"网络;运用R软件进行GO和KEGG分析;使用Autodock Vina和Py-mol软件进行平消胶囊活性成分与核心靶点的分子对接及可视化;通过R软件survival包对核心靶点进行分析,筛选出与总体生存时间密切相关的基因;通过CCK-8法检测细胞活力;流式细胞术检测细胞凋亡;Western blot检测 p-AKT1、β-catenin和cyclinD1的蛋白表达水平.结果:筛选出药物靶点194个,疾病靶点1 612个,韦恩图求交集靶点共127个,核心靶点主要包括TB53、AKT1、TNF、CASP3等20个;GO分析主要与氧化应激反应、细胞对化学反应的调控等生物活性有关;KEGG分析主要通过PI3K-AKT信号通路、TNF信号通路、IL-17信号通路等通路有关;分子对接结果显示活性成分与核心靶点AKT1、MAPK1、RELA均有较好的结合;细胞实验表明槲皮素(40、80、120 μmol/L)促进乳腺癌细胞凋亡;Western blot检测显示随着不同浓度的槲皮素处理乳腺癌细胞48 h后,p-AKT1、β-catenin和cyclinD1的蛋白表达量均不同程度地降低.结论:网络药理学与细胞实验证实平消胶囊可能通过调节AKT1/β-catenin信号通路,发挥其抗乳腺癌作用.
Abstract
AIM:To explore the potential mecha-nism of action of Pingxiao capsule in the treatment of breast cancer.METHODS:TCMSP,TCM-ID,Gene-Cards and other databases were used to screen the related targets of Pingxiao capsule and breast can-cer.Cytoscape software builds drug-target-disease networks.R software was used for GO and KEGG analysis.Autodock Vina and Pymol software were used for molecular docking and visualization of Pingxiao capsule active ingredients and core tar-gets.Core targets were analyzed by R software sur-vival package,and genes closely related to overall survival time were screened out.Cell viability was detected by CCK-8 method.Flow cytometry was used to detect cell apoptosis.The protein expres-sion levels of p-AKT1,β-catenin and cyclinD1 were detected by Western blot.RESULTS:A total of 194 drug targets were screened,1612 disease targets were identified,127 intersection targets were iden-tified by Venn diagram,and 20 core targets were TB53,AKT1,TNF,CASP3,etc.GO analysis was main-ly related to oxidative stress response,cell regula-tion of chemical reaction and other biological activi-ties.KEGG analysis was mainly related to PI3K-AKT signaling pathway,TNF signaling pathway,IL-17 sig-naling pathway and other pathways.Molecular docking results showed that the active constituents were well combined with the core targets AKT1,MAPK1 and RELA.Cell experiments showed that quercetin(40,80,120 μmol/L)promoted apoptosis of breast cancer cells.Western blot analysis showed that the protein expressions of p-AKT1,β-catenin and cyclinD1 decreased with different con-centrations of quercetin treated for 48h.CONCLU-SION:Network pharmacology and cell experiments confirmed that Pingxiao capsule may exert its anti-breast cancer effect by regulating AKT1/β-catenin signaling pathway.
基金项目
安徽省自然科学基金(2208085MH252)
安徽省重点研究与开发计划项目(201904a07020092)
安徽省自然科学基金(2108085MH311)
国家科技期刊平台
NSTL
维普
万方数据