基于网络药理学和分子实验探讨蝙蝠葛碱对鼻咽癌的作用机制

The mechanism of dauricine on nasopharyngeal carcinoma based on network pharmacology and molecular experiment

目的:通过网络药理学分析蝙蝠葛碱对鼻咽癌(NPC)的作用靶点及通路,并探讨其作用机制。方法:运用Pubchem、Pharmmapper、GeneCards数据库检索蝙蝠葛碱和NPC的作用靶点后并取得它们的交集靶点;使用Cytoscape软件构建“蝙蝠葛碱—NPC疾病靶点”网络图,并利用David数据库进行GO功能注释及KEGG通路富集分析。CCK-8法检测蝙蝠葛碱作用的CNE-2细胞在24 h、48 h、72 h的增殖抑制情况。Western blotting法检测预测出的通路靶点蛋白的表达。结果:共筛选出潜在作用靶点69个,GO功能注释获得分子功能条目16个、生物过程条目24个和细胞组成条目10个,KEGG通路富集得到20条相关通路,涉及PI3K-AKT信号、癌症信号、催乳激素等信号通路。蝙蝠葛碱呈浓度和时间依赖性抑制CNE-2细胞增殖,PI3K和AKT蛋白表达明显下调(P<0.05)。结论:蝙蝠葛碱能有效抑制NPC细胞增殖,并且验证了PI3K-AKT通路为蝙蝠葛碱作用于NPC的主要通路。

Objective:To analyze the target and pathway of dauricine on nasopharyngeal carcinoma(NPC) by network pharmacology,and to explore its mechanism.Methods:The argets of dauricine and NPC were searched by Pubchem,Pharmmapper and GeneCards databases,and their intersection targets were obtained.The network map of "dauricine-NPC disease targets" was constructed by Cytoscape software,and the GO function annotation and KEGG pathway enrichment analysis were carried out by David database.The proliferation and inhibition of CNE-2 cells treated with dauricine at 24 h,48 h and 72 h were detected by CCK-8.The expression of predicted pathway target protein was detected by Western blotting.Results:A total of 69 potential targets were screened,16 molecular function items,24 biological process items and 10 cell composition items were obtained by GO functional annotation,and 20 related pathways were obtained by enrichment of KEGG pathway,involving PI3KAKT signal,cancer signal,prolactin and other signal pathways.Dauricine inhibited the proliferation of CNE-2cells in a concentration-and time-dependent manner,and the expressions of PI3K and AKT proteins were significantly down-regulated(P<0.05).Conclusion:Dauricine can effectively inhibit the proliferation of NPC cells,and it is verified that PI3K-AKT pathway is the main pathway of dauricine on NPC.