高通量测序分析阿尔茨海默病患者血清circRNA表达谱的研究  

Research of circRNA Expression in Serum of Patients with Alzheimer Disease by High-Throughput Sequencing

作　　者：郭清允 崔健 吴睿[1] GUO Qingyun;CUI Jian;WU Rui(Department of Neurological Rehabilitation,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou,450003)

机构地区：[1]郑州大学第二附属医院神经康复科,河南郑州450000

出　　处：《河南医学研究》2025年第4期598-603,共6页Henan Medical Research

摘　　要：目的应用高通量测序分析阿尔茨海默病(AD)患者血清circRNA的表达谱差异,结合临床诊断,筛选可辅助诊断AD的分子标志物。方法收集AD患者(AD组)及健康人群(对照组)各28例,用高通量测序将AD组和对照组的全血circRNA进行检测,筛选出差异表达的circRNA,通过基因本体(GO)富集分析和信号通路的显著性(KEGG)分析选出AD差异基因,通过生物信息学分析检测出表达显著升高的circ_0089972与miR-7,并找出其内在关系,采用双荧光素酶法验证。结果AD患者的血清中有显著性差异表达的circRNA有29个,其中表达上调的20个,表达下调的9个(P<0.05),GO富集分析结果显示,差异表达的circRNA主要富集在细胞代谢过程、蛋白结合、有机物代谢过程、细胞活动调节、氮化合物代谢过程、细胞对刺激反应、有机环化物的结合、离子结合、代谢过程调节、细胞通信、信号传递等生物学过程中。而KEGG分析显示差异基因主要参与癌症通路、PI3K-Akt信号通路、人乳头状瘤病毒感染、微小RNA、MAPK信号通路、Ras信号通路、EB病毒感染等信号通路的调节。生物信息学及双荧光素酶分析显示circ_0089972可作为分子海绵(可与RNA分子相互作用,竞争性吸附microRNA,调节miRNA的水平和/或活性)吸附并下调miR-7表达,从而参与AD的病理生理过程。结论AD患者血清中存在明显差异表达的circRNA,差异表达的circRNA通过其靶基因参与多个AD生物学过程和信号通路调节,其中hsa_circ_0089972等可能成为辅助诊断AD的分子标志物。Objective To detect the difference of circRNA expression profile in the serum of Alzheimer disease(AD)patients by high-throughput sequencing and find the potential diagnostic biomarkers of AD in combination with clinical diagnosis.Methods There were 28 AD patients(AD group)and 28 healthy individuals(normal control group)were recruited.High throughput sequencing was used to test whole blood circRNA and screen differential expressed circRNA.The gene ontology database enrichment analysis(GO enrichment analysis)and signal pathway significance analysis(KEGG analysis)of signaling pathways choose the genetic variations of AD were used to analyze the differentially expressed circRNAs.Differential genes of AD were analyzed and selected,and circ 0089972 and miR-7 with significantly increased expression were detected through bioinformatics analysis,and their internal relationship was found out,and verified by double luciferase method.Results This study found that there were 29 circrnas with significant differentially expressed expression in the serum of AD patients,of which 20 were up-regulated and 9 were down-regulated(P<0.05).GO enrichment analysis results show that the circRNA main enrichment of differentially expressed in cell metabolism,protein,organic metabolism,cell activity regulation,nitrogen metabolism,cell response to stimuli,organic combination of ring compound,ion combination,metabolism regulation,cell communication,signal transmission and other biological process.KEGG analysis showed that differential genes were mainly involved in the regulation of cancer pathways,PI3K-Akt signaling pathway,human papilloma virus infection,microRNA,MAPK signaling pathway,Ras signaling pathway,EB virus infection and other signaling pathways.The result of bioinformatics and dual-luciferase assay showed that circ_0089972 could be used as a molecular sponge(can interact with RNA molecules,competitively adsorb micrornas,and regulate miRNA levels and/or activity)to adsorb and down-regulate the expression of miR-7,thus participating i

关 键 词：阿尔茨海默病 circRNA 高通量测序 差异基因 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]