Abstract: Objective To explore the mechanism of cordycepin (Cor) in the treatment of non-small cell lung cancer ( NSCLC ) by network pharmacology, molecular docking and in vitro experiments. Methods Multiple databases were used to search the targets of Cor and the therapeutic targets of NSCLC, and the common targets were screened out. STRING database and Cytostape were used to analyze and visualize the protein-protein interaction networks (PPI). GO and KEGG enrichment analysis was performed using the DAVID database, and the results were visualized using the online tool “Weishengxin”. AutoDock Vina was used to verify the molecular docking of Cor with the core target, and the binding energy was calculated. PyMol was used to visualize the results. MTT assay was used to detect the effect of Cor on the proliferation of NSCLC A549 cells. Western blot was used to detect the effect of Cor on the expression of core target proteins PARP1, P53 and AKT1. Results A total of 215 targets of Cor and 1238 therapeutic targets of NSCLC were screened, and 122 common targets were obtained. After PPI analysis, the core targets PARP1, P53 and AKT1 were selected according to Degree value for verification. The results of GO and KEGG enrichment analysis showed that biological process (BP) mainly involved apoptosis process, positive and negative regulation of gene expression, cellular component ( CC ) mainly involved cytoplasm and nucleus, molecular function (MF) mainly involved protein binding and enzyme binding, and signaling pathways mainly involved cancer, PI3K / AKT and other signaling pathways. Molecular docking showed that the three targets of PARP1, P53 and AKT1 were highly correlated with the treatment of NSCLC. In vitro experiments showed that Cor could inhibit the proliferation of human NSCLC A549 cells, significantly up-regulate the expression of P53 and PARP1 and down-regulate the expression of AKT1. Conclusion Cor inhibits the proliferation of human NSCLC A549 cells, and the mechanism may be related to the regulation of PARP1, P53 and AKT1 expression.

Key words: Cordycepin, non-small cell lung cancer, network pharmacology, molecular docking

摘要： 目的 通过网络药理学、分子对接和体外实验探讨虫草素（Cor）治疗非小细胞肺癌（NSCLC）的作用机制。方法 采用多个数据库检索Cor的作用靶点和NSCLC的治疗靶点,筛选出共同作用靶点,并采用STRING数据库和Cytostape对其进行蛋白–蛋白相互作用（PPI）分析和可视化处理。采用DAVID数据库对其进行GO和KEGG富集分析,使用线上工具“微生信”对结果进行可视化处理。采用AutoDock Vina进行Cor与核心靶点的分子对接,计算结合能,使用PyMol对结果进行可视化处理。采用MTT法检测Cor对NSCLC A549细胞增殖作用的影响,采用Western blot检测Cor对核心靶点蛋白PARP1、P53、AKT1表达情况的影响。结果 筛选获得Cor的作用靶点215个、NSCLC的治疗靶点1238个,获得122个共同作用靶点,进行PPI分析后,根据Degree值选择核心靶点PARP1、P53、AKT1用于验证。GO和KEGG富集分析结果显示,生物过程（BP）主要涉及凋亡过程、基因表达的正、负调控等,细胞组成（CC）主要涉及细胞质、细胞核等,分子功能（MF）主要涉及蛋白结合、酶结合等,信号通路主要涉及癌症、PI3K/AKT等。分子对接显示,PARP1、P53、AKT1三个靶点与NSCLC的治疗呈现高度相关性。体外实验显示,Cor可抑制人NSCLC A549细胞增殖,并显著上调P53和PARP1的表达,同时下调AKT1的表达。结论 Cor抑制人NSCLC A549细胞的增殖,机制可能与调节PARP1、P53、AKT1表达有关。

关键词: 虫草素, 非小细胞肺癌, 网络药理学, 分子对接