Erastin治疗肺癌的网络药理学分析和体外实验验证

Effects and Mechanisms of Erastin in Treatment of Lung Cancer Based on Network Phatmacology and Verified Experiments in Vitro

目的本研究通过网络药理学分析小分子化合物Erastin治疗肺癌的生物学信息,并通过细胞实验进行体外验证。方法通过数据库筛选获取Erastin的主要靶点及肺癌的疾病作用靶点。经过韦恩(Venn)图解析获取Erastin和肺癌的共同靶点,然后构建蛋白-蛋白质相互作用(PPI)网络、活性成分-靶点网络及疾病-药物-活性成分-靶点网络。选择不同浓度Erastin干预小鼠Lewis肺癌细胞(LLC细胞),通过细胞增殖-毒性实验(CCK-8)检测细胞的增殖抑制率,四唑盐比色法(MTT)检测细胞的存活率,平板克隆检测细胞的增殖活力,Transwell实验和划痕实验分别检测LLC细胞的侵袭和迁移能力。结果筛选得到184个与Erastin活性成分关联的靶点,4197个与肺癌相关靶点,其中有98个共同靶点,这些靶标与蛋白质磷酸化、蛋白质结合、胞质密切相关。体外实验证实,随着Erastin浓度的增加,LLC细胞的增殖抑制率逐渐上升,存活率逐渐下降,迁移和侵袭的细胞数逐渐减少。结论Erastin对LLC细胞有明显的抑制作用,其作用可能与两者的多个共同靶点有关。

Objective In this study,the biological information of the small-molecule compound erastin in the treatment of lung cancer was analyzed by network pharmacology and verified by cell experiments in vitro.Methods The main targets of erastin and lung cancer were obtained through database screening.The common targets of erastin and lung cancer were obtained by Venn diagram,and then protein-protein interaction(PPI)network,active ingredient-target network and disease-drug-active ingredient-target network were constructed.Mouse Lewis lung cancer(LLC)cells were treated with different concentrations of erastin,and cell proliferation inhibition rate was detected by cell counting kit-8(CCK-8),cell viability rate was detected bymethyl thiazolyl tetrazolium(MTT),cell proliferation viability rate was detected by colony formation assay,invasive and migratory ability of LLC cells were detected by Transwell assay and Wound-Healing assay,respectively.Results About 184 active ingredient-associated targets of erastin and 4197 targets associated with lung cancer were screened,and showed 98 common targets,which were closely related to protein phosphorylation,protein binding and cytoplasmic.In vitro experiments of LLC cells confirmed that with the increase of erastin concentration,the proliferation inhibition rate gradually increased,the survival rate gradually decreased,and the number of migrating and invading cells gradually decreased.Conclusion Erastin has a significant inhibitory effect on LLC cells,and this effect may be related to their multiple common targets.