基于网络药理学、分子对接和体外实验探讨补骨脂定抗鼻咽癌的作用机制

Exploration of the effect and mechanism of psoralidin against nasopharyngeal carcinoma based on network pharmacology,molecular docking and in vitro experiments

目的:基于网络药理学和生物信息学探讨补骨脂定(PSO)治疗鼻咽癌的作用及分子机制。方法:将鼻咽癌5-8F细胞分为5-8F组(0μmol/L PSO),PSO低剂量组(10μmol/L组)、中剂量组(20μmol/L组)、高剂量组(30μmol/L组)。通过CCK-8、平板克隆探究PSO对鼻咽癌5-8F细胞增殖、克隆形成的影响;Pubchem、Pharmmapper、GeneCards数据库得到PSO治疗鼻咽癌的潜在靶点;DAVID数据库进行GO、KEGG分析;STRING、GEPIA数据库和Cytoscape软件构建网络图并得到核心靶点;分子对接、western blotting对核心靶点及主要通路进行验证。结果:PSO呈浓度依赖性抑制5-8F细胞的增殖;与5-8F组比较,PSO各剂量组细胞克隆形成能力降低(P<0.05);PSO治疗鼻咽癌的潜在靶点共有66个,KEGG分析显示,与PI3K-Akt信号通路密切相关;核心靶点为ALB、HSP90AA1、SRC、EGFR、CASP3、ANXA5、MAPK1,其中ALB、HSP90AA1、ANXA5为生存相关靶点;PSO与核心靶点ALB、HSP90AA1、ANXA5具有良好的结合能力;与5-8F组比较,PSO高剂量组细胞中PI3K、p-PI3K、Akt、pAkt蛋白水平的表达降低(P<0.05)。结论:PSO可有效抑制鼻咽癌5-8F细胞的增殖、克隆形成,其抗鼻咽癌机制主要与抑制PI3K-Akt信号通路活性有关。

Objective:To explore the effect and molecular mechanism of psoralidin(PSO)in the treatment of nasopharyngeal carcinoma based on network pharmacology and bioinformatics.Methods:The nasopharyngeal carcinoma 5-8F cells were divided into 5-8F group(0μmol/L PSO),low-dose PSO group(10μmol/L group),middle-dose PSO group(20μmol/L group)and high-dose PSO group(30μmol/L group).The effect of PSO on the proliferation and clone formation of nasopharyngeal carcinoma 5-8F cells was explored by CCK-8 and plate cloning.Pubchem,Pharmmapper and GeneCards databases got the potential target of PSO in the treatment of nasopharyngeal carcinoma;DAVID database carried out GO and KEGG analysis;STRING,GEPIA databases and Cytoscape software constructed a network diagram and got the core target;molecular docking and western blotting were used to verify the core targets and main pathways.Results:PSO inhibited the proliferation of 5-8F cells in a concentration-dependent manner.Compared with 5-8F group,the ability of cell clone formation in each dose group of PSO decreased(P<0.05).There were 66 potential targets of PSO in the treatment of nasopharyngeal carcinoma,and KEGG analysis shows that it was closely related to PI3K-Akt signal pathway.The core targets were ALB,HSP90AA1,SRC,EGFR,CASP3,ANXA5 and MAPK1,among which ALB,HSP90AA1 and ANXA5 were survival-related targets;PSO had good binding ability with the core targets ALB,HSP90AA1 and ANXA5.Compared with the 5-8F group,the expressions of PI3K,p-PI3K,Akt and p-Akt protein in the highdose PSO group decreased(P<0.05).Conclusion:PSO can effectively inhibit the proliferation and clone formation of nasopharyngeal carcinoma 5-8F cells,and its anti-nasopharyngeal carcinoma mechanism is mainly related to the inhibition of PI3K-Akt signal pathway activity.