基于生物信息学和体外实验探讨蝙蝠葛碱治疗肺腺癌的靶点及机制  

Investigating the Targets and Mechanisms of Dauricine in the Treatment of Lung Adenocarcinoma through Bioinformatics and In Vitro Experiments

作　　者：程龙 熊媛 杨慧 钱铭 葛卫红[1] Cheng Long;Xiong Yuan;Yang Hui;Qian Ming;Ge Weihong(Department of Pharmacy,Nanjing Drum Tower Hospital(i.e.Clinical College of Nanjing University of Chinese Medicine),Nanjing 210008,Jiangsu,China;School of Basic Medicine and Clinical Pharmacy,China Pharmaceutical University,Nanjing 210009,Jiangsu,China)

机构地区：[1]南京中医药大学鼓楼临床医学院/南京鼓楼医院药学部,南京210008 [2]中国药科大学基础医学与临床药学学院,南京210009

出　　处：《肿瘤预防与治疗》2025年第3期171-181,共11页Journal of Cancer Control And Treatment

基　　金：国家自然科学基金项目(编号:82204720)。

摘　　要：目的:结合生物信息学和体外实验阐明蝙蝠葛碱(dauricine,DAU)治疗肺腺癌(lung adenocarcinoma,LUAD)的作用和机制。方法:从SwissTargetPrediction数据库预测DAU作用靶点,利用TCGA数据库检索LUAD的相关靶点,并进行差异表达分析;将DAU和LUAD的差异表达基因(differentially expressed genes,DEGs)进行交叉筛选,从而获得交集基因,构建蛋白质-蛋白质相互作用网络;通过DAVID数据库利用基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)对交集基因进行功能富集分析,通过Sangerbox进行患者生存分析。体外培养A549和NCI-H1975肺腺癌细胞,MTS法检测不同浓度DAU对LUAD细胞增殖的影响;利用RT-qPCR检测DAU对预测靶基因mRNA表达水平的影响。结果:筛选获得270个DAU相关靶点,KEGG分析显示DAU靶点主要富集在肺癌。获得5367个LUAD的DEGs,二者共有23个交集基因,KEGG功能富集分析显示,靶基因主要富集在细胞增殖、神经活性配体-受体相互作用、癌症、多巴胺能突触、5-羟色胺能突触等相关生物学过程及通路。生存分析显示,MELK、HTR3A、AVPR1B、ADRB2、SLC6A3、SELP、ACHE、MTNR1A、RET、OPRD1基因与LUAD患者生存率有关。体外实验结果显示,DAU显著抑制LUAD细胞的增殖。DAU升高ADRA1A、ACHE、SLC6A4、CFD、TERT、EZH2的mRNA表达水平,降低MELK、AURKA、BCHE的mRNA表达水平。结论:本研究发现DAU具有抑制LUAD细胞增殖的作用,并结合生物信息学和体外实验分析验证DAU治疗LUAD的潜在靶点和机制,为DAU应用于LUAD的治疗提供了参考依据。Objective:To explore dauricine(DAU)'s therapeutic effects and mechanisms in lung adenocarcinoma(LUAD)through bioinformatics and in vitro studies.Methods:Potential targets of DAU were predicted using the SwissTargetPrediction database,while LUAD-related targets were obtained from The Cancer Genome Atlas database for differential expression analysis.Overlapping genes between DAU and LUAD differentially expressed genes(DEGs)were identified and used to construct a protein-protein interaction network.Functional enrichment analysis of the overlapping genes was conducted using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses through the DAVID database,while patient survival analysis was performed via Sangerbox.A549 and NCI-H1975 cells were cultured in vitro,and the effects of different concentrations of DAU on cell proliferation were assessed using the MTS assay.RT-qPCR was performed to analyze the effects of DAU on the mRNA expression levels of predicted target genes.Results:A total of 270 DAU-related targets were identified,and KEGG pathway analysis demonstrated significant enrichment of these targets in lung cancer-related pathways.A total of 5,367 DEGs were identified in LUAD,with 23 intersecting genes shared between DAU and LUAD.KEGG functional enrichment analysis revealed that the target genes were predominantly enriched in biological processes and pathways,including cell proliferation,neuroactive ligand-receptor interactions,cancer,dopaminergic synapses,5-hydroxytryptaminergic synapses,and others.Survival analysis revealed that the genes MELK,HTR3A,AVPR1B,ADRB2,SLC6A3,SELP,ACHE,MTNR1A,RET,and OPRD1 are associated with the survival rate of LUAD patients.The results of the in vitro experiment demonstrated that DAU significantly inhibited the proliferation of LUAD cells.DAU upregulates the mRNA expression levels of ADRA1A,ACHE,SLC6A4,CFD,TERT,and EZH2,while downregulating the mRNA expression levels of MELK,AURKA,and BCHE.Conclusion:This study demonstrated that DAU inhibits the prolifer

关 键 词：蝙蝠葛碱 肺腺癌 生物信息学 

分 类 号：R979.1[医药卫生—药品]