摘要
目的基于系统生物信息学分析鼻咽癌样本基因芯片数据,筛选鼻咽癌的潜在预测基因,并对其进行实验验证。方法通过基因表达综合数据库筛选鼻咽癌数据集,通过limma包进行差异分析,加权基因共表达网络包进行加权基因共表达网络分析,绘制韦恩图寻找共同基因,进行基因本体富集分析及京都基因与基因组百科全书通路分析。通过蛋白质互作网络、LASSO回归及非参数检验,进一步挖掘鼻咽癌的生物标志物。通过实时荧光定量PCR和Western blot对鼻咽癌关键预测基因的mRNA及蛋白表达情况进行检测,验证结果。结果GSE12452数据集上调基因共622个,下调基因共351个,通过limma分析及加权基因共表达网络分析并交集最终获取116个交集基因,通过富集分析,生物过程主要包括细胞增殖及调控、细胞间黏附的调节等,富集于Rap1信号通路、Ras信号通路、肿瘤坏死因子信号通路等通路,经过筛选得到血管生成素2(ANGPT2)、双氧化酶2(DUOX2)、人组织因子Ⅲ(F3)、白细胞介素-15(IL-15)、脂质运载蛋白2、视黄酸受体相关孤儿受体B(RORB)共6个关键基因,实时荧光定量PCR和Western blot实验证明ANGPT2、IL-15的mRNA和蛋白在鼻咽癌细胞中高表达,DUOX2、F3、RORB的mRNA和蛋白低表达。结论ANGPT2、DUOX2、F3、IL-15、RORB为鼻咽癌的潜在预测分子标志物和治疗靶点,其机制可能与Rap1、Ras、肿瘤坏死因子等信号通路有关。
Objective To screen out the potential prediction genes for nasopharyngeal carcinoma(NPC)from the gene microarray data of NPC samples and then verify the genes by cell experiments.Methods The NPC dataset was downloaded from Gene Expression Omnibus,and limma package was employed to screen out the differentially expressed genes.Weighted correlation network analysis package was used for weighted gene co-expression network analysis,and Venn diagram was drawn to find the common genes.The gene ontology annotation and Kyoto encyclopedia of genes and genomes pathway enrichment were then performed for the common genes.The biomarkers for NPC were further explored by protein-protein interaction network,LASSO regression,and non-parametric tests.Real-time quantitative PCR and Western blotting were employed to determine the mRNA and protein levels of key predictors of NPC,so as to verify the screening results.Results There were 622 up-regulated genes and 351 down-regulated genes in the GSE12452 dataset.A total of 116 common genes were obtained by limma analysis and weighted gene co-expression network analysis.The common genes were mainly involved in the biological processes of cell proliferation and regulation and regulation of intercellular adhesion.They were mainly enriched in Rap1,Ras,and tumor necrosis factor signaling pathways.Six key genes were screened out,encoding angiopoietin-2(ANGPT2),dual oxidase 2(DUOX2),coagulation factorⅢ(F3),interleukin-15(IL-15),lipocalin-2,and retinoic acid receptor-related orphan receptor B(RORB).Real-time quantitative PCR and Western blotting showed that the NPC cells had up-regulated mRNA and protein levels of ANGPT2 and IL-15 and down-regulated mRNA and protein levels of DUOX2,F3,and RORB,which was consistent with the results predicted by bioinformatics.Conclusion ANGPT2,DUOX2,F3,IL-15 and RORB are potential predictive molecular markers and therapeutic targets for NPC,which may be involved in Rap1,Ras,tumor necrosis factor and other signaling pathways.
作者
黄子葵
李斐
陈跃洲
刘佳铭
刘嘉俊
蓝素珍
HUANG Zikui;LI Fei;CHEN Yuezhou;LIU Jiaming;LIU Jiajun;LAN Suzhen(Department of Traditional Chinese Medicine,Ganzhou People’s Hospital,Ganzhou,Jiangxi 341000,China;Shenzhen Clinical School of Medicine,Guangzhou University of Chinese Medicine,Shenzhen,Guangdong 518116,China;The Second Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Clinical Medical College of Acupuncture Moxibustion and Rehabilitation,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Department of Oncology,Ganzhou People’s Hospital,Ganzhou,Jiangxi 341000,China)
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2023年第4期597-607,共11页
Acta Academiae Medicinae Sinicae
基金
江西省卫生健康委员会中医药科研课题(2018B109)。
作者简介
通信作者:蓝素珍,电话:0797-5889396,电子邮件:l18170115016@163.com。
