摘要
利用高通量测序筛选心房颤动(房颤)伴脑卒中患者外周血单核细胞中差异表达的lncRNAs,为从分子水平探讨房颤脑卒中发生发展机制提供实验依据,并寻找预测房颤脑卒中相关生物学标志物.应用高通量测序检测三例房颤伴脑卒中患者和三例房颤患者外周血单核细胞lncRNAs差异表达,进行基因本体论(Gene Ontology,GO)、代谢信号通路(Pathway)分析,建立lncRNAs-miRNAs共表达网络,并进行实时荧光定量PCR(qRT-PCR)验证.与对照组相比,房颤伴脑卒中患者差异表达301条lncRNA,其中上调143条,下调158条,上调和下调表达差异最大的是LINC01002,HCG18等基因.GO功能分析发现,差异表达基因参与疾病发生发展最主要的生物学过程包括结合、肽交联、高渗反应、内肽酶活性的调节、细胞体积的正向调节等.KEGG分析后发现,差异表达的lncRNA相关信号通路主要涉及免疫系统和炎症等相关.TCONS_00014860,TCONS_00020132,TCONS_00037601,TCONS_00003375,TCONS_00013755是lncRNAs-miRNAs共表达网络中结合节点最多的前五个lncRNAs,hsa-miR-619-5p是共表达网络中与lncRNAs作用最多的节点.qRT-PCR验证LINC01002,HCG18在两组人群中差异表达具有统计学意义.其与房颤相比,房颤伴脑卒中患者外周血存在差异表达lncRNA,与房颤发生脑卒中相关.XLOC_005045,XLOC.006945,XLOC_013107,XLOC.001139和XLOC_004677是共表达网络中结合节点最多的前五个lncRNAs,lncRNA-hsa-miR-619-5p可能在房颤伴脑卒中的发病机制中起到重要作用.同时,LINC01002,HCG18可以作为预测持续性房颤合并卒中的新型生物学标志物.
Using microarray to detect differentially expressed long non-coding RNAs(lncRNAs)in human Peripheral blood mononuclear cells(PBMC)from Atrial Fibrillation(AF)-related stroke patients and AF controls,we explore a theoretical basis for the mechanism of persistent AF~related stroke and search for novel biological markers.The peripheral blood monocytes of three patients with AF-related stroke and three AF subjects are collected for transcriptome sequencing(RNA-seq)to detect differently expressed lncRNAs.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis are performed to identify the functions of differently expressed genes and related pathways.Then We verify the expression of LINC01002 and HCG18 through qRT-PCR.There are 301 differently expressed lncRNAs,including 143 up-regulated and 158 down-regulated expressions respectively.Besides,the most up-regulated and down-regulated expressions are LINC01002 and HCG18.GO analysis shows that differentially expressed genes enriched in differentially expressed transcripts in the biological process are mainly involved in copulation,peptide cross-linking,hyperosmotic response,regulation of endopeptidase activity,and positive regulation of cell volume process.KEGG enrichment pathway analysis shows that some of the enrichment pathways associate with differentially expressed lncRNAs include immune system and infectious diseases,etc.Network analysis uncover five key lncRNAs,which are XLOC.005045,XLOC_006945,XLOC_013107,XLOC_001139,and XLOC.004677.Hsa-miR-61-5p is the highest positive correlated miRNA in the networks.The results of qRT PCR show that the expression of LINC01002 and HCG18 in the two groups are significantly different.There are differentially expressed lncRNAs in human monocytes from AF-related stroke patients.The lncRNAs-hsa-miR-619-5p-mediated network may play a critical role in the pathogenesis of AF-related stroke.LINC01002 and HCG18 can be novel biological markers for predicting AF-related stroke.
作者
崔进进
陈茜
丛树一
阮中宝
Cui Jinjin;Chen Xi;Cong Shuyi;Ruan Zhongbao(Taizhou People's Hospital Affiliated to Nanjing University of Chinese Medicine,Taizhou,225300,China;Department of Cardiology,Jiangsu Taizhou People’s Hospital,Taizhou,225300,China)
出处
《南京大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第1期175-182,共8页
Journal of Nanjing University（Natural Science）
基金
泰州市科技支撑计划。
