蛇葡萄素抗乳腺癌的网络药理学分析及细胞试验研究

Network Pharmacological Analysis and Cell Test Research of Ampelopsin Against Breast Cancer

目的探讨蛇葡萄素(AMP)抗乳腺癌的药理学作用机制。方法通过Pubchem数据库获取AMP3D分子结构,上传至PharmMapper数据库,获得AMP作用靶点基因,利用Genecards数据库获取乳腺癌疾病靶点基因;通过Venny R软件获得AMP与乳腺癌作用靶点交集,并输入String数据库,以Cytoscape3.2.1软件构建"药物-靶点-疾病"可视化网络;再进行GO和KEGG富集分析。体外培养MDA-MB-231细胞,通过CCK-8试验、划痕试验、迁移及逆转录聚合酶链反应进行验证。结果共选出119个靶基因;GO富集分析显示,靶基因主要集中在氧化应激等生物学过程;KEGG通路富集分析显示,AMP抗乳腺癌作用靶基因涉及PI3K-Akt,MAPK,Ras等信号通路。体外验证试验结果显示,AMP可明显抑制MDA-MB-231细胞增殖和迁移,下调关键靶基因非受体酪氨酸激酶和基质金属蛋白酶2的表达。结论该研究为AMP用于乳腺癌的防治提供了理论依据和研发思路。

Objective To investigate the pharmacological mechanism of ampelopsin(AMP)in the treatment of breast cancer.Methods AMP 3 D molecular structure was obtained from PubChem database and uploaded to PharMapper to obtain AMP target genes.The breast cancer target genes were obtained by Genecards database.The intersection of AMP and breast cancer target was obtained by Venny R software and input into the String database to construct the″drug-target-disease″visual network by Cytoscape 3.2.1 software.GO and KEGG enrichment analysis was performed.MDA-MB-231 cells were cultured in vitro and verified by CCK-8,scratch,migration and RT-qPCR.Results A total of 119 target genes were screened.GO enrichment analysis showed that the target genes were mainly concentrated in biological processes such as oxidative stress,and KEGG enrichment analysis showed that the target genes of AMP anti-breast cancer were involved in PI3 K-Akt,MAPK,Ras and other signaling pathways.The results of in vitro validation test showed that AMP could significantly inhibite the proliferation and migration of MDA-MB-231 cells,and down-regulate the expression of key target genes,nonreceptor tyrosine kinase(SRC)and matrix metalloproteinase-2(MMP2).Conclusion This study provides a theoretical basis and the research ideas for the application of AMP in the prevention and treatment of breast cancer.