基于网络药理学分析瑞香素抗多种肿瘤的作用机制及体外实验验证  

Study on the anti-tumor mechanism of daphnetin based on network pharmacology analysis and experimental verification in vitro

作　　者：冯松 张小东[2] 覃亚亚 彭彬[1] FENG Song;ZHANG Xiao-dong;QIN Ya-ya;PENG Bin(North Sichuan Medical College;Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院 [2]川北医学院附属医院,南充637000

出　　处：《天然产物研究与开发》2022年第1期144-152,共9页Natural Product Research and Development

基　　金：川北医学院国家预科项目(CBY19-YZ12);南充市学校科技战略合作(19SXHZ0106);川北医学院附属医院研发项目(2020ZD003)。

摘　　要：基于网络药理学预测瑞香素抗恶性胶质瘤、肝癌和三阴性乳腺癌的共同靶点及可能机制,并对其进行体外实验验证。利用Swiss Target Prediction和GeneCards等数据库检索瑞香素与恶性胶质瘤、肝癌和三阴性乳腺癌的共同靶点。使用Cytoscape构建瑞香素-三种肿瘤蛋白质相互作用网络图(PPI)并筛选出核心靶点,并对核心靶点进行GO及KEGG富集分析;通过AutoDock Tools对瑞香素与核心靶点进行分子对接。体外实验验证:采用CCK-8和Western blot法行体外实验验证不同浓度瑞香素对U-251 MG、HepG-2和MDA-MB231细胞系细胞抑制率和P53、RRM2蛋白的表达水平的影响。共筛选出瑞香素抗三种肿瘤核心靶点56个,富集分析显示靶点富集在P53通路和癌症通路,参与细胞周期调节、细胞凋亡、DNA生物合成和修复等生物过程;分子对接结果显示瑞香素与P53、RRM2有较好的结合作用。体外验证实验显示,与对照组比较,瑞香素能显著抑制U-251 MG、HepG-2、MDA-MB231的增殖(P<0.01),并显著上调其P53蛋白及下调RRM2蛋白的表达,且呈剂量依赖性(P<0.05)。我们的结果提示瑞香素能抑制U-251 MG、HepG-2和MDA-MB231细胞增殖,其机制可能与调控P53/RRM2通路有关。In this study,network pharmacology prediction and in vitro experimental verification were used to explore the common targets and possible mechanisms of daphnetin against malignant glioma,liver cancer and triple-negative breast cancer.The targets of daphnetin and malignant glioma,hepatocellular carcinoma and triple negative breast cancer were retrieved by using Swiss Target Prediction and GeneCards database,respectively.The protein-protein interaction network between daphnetin and three kinds of tumors was constructed by using Cytoscape,and the core targets were screened.GO and KEGG enrichment analysis was performed on the core targets.The molecular docking study of daphnetin with core targets were carried out using the AutoDock Tools.CCK-8 and Western blot were used to verify the effects of daphnetin on the cell inhibition rate and the protein expression levels of p53 and RRM2 in U-251MG,HepG-2 and MDA-MB231 cell lines.56 core targets of daphnetin against three kinds of tumor were screened.Enrichment analysis showed that the core targets were enriched in p53 pathway and cancer pathway and involved in biological processes such as cell cycle regulation,apoptosis,DNA biosynthesis and repair.Molecular docking results showed that daphnetin strongly bound with P53 and RRM2.In vitro results showed that Daphnetin significantly inhibited U-251 MG,HepG-2,MDA-MB231 proliferation(P<0.01),up-regulated P53 protein expression and down-regulated RRM2 expression in a dose-dependent manner(P<0.05).Our results suggested that daphnetin can inhibit the proliferation of U-251MG,HepG-2 and MDA-MB231 cells,and its mechanism may be related to the regulation of p53/RRM2 pathway.

关 键 词：瑞香素 网络药理学 恶性胶质瘤 肝癌 三阴性乳腺癌 

分 类 号：R285.5[医药卫生—中药学]