AGPS基因沉默对神经胶质瘤细胞lncRNA、mRNA表达谱的影响及意义  

Effects of AGPS gene silencing on lncRNA and mRNA expression profiles in glioma cells and their significance

作　　者：崔菀真 朱彧[2] 岳伟[2] CUI Wanzhen1];ZHU Yu;YUE Wei(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;不详)

机构地区：[1]天津中医药大学,天津301617 [2]天津市环湖医院天津市脑血管与神经变性重点实验室

出　　处：《山东医药》2019年第35期6-11,共6页Shandong Medical Journal

基　　金：国家自然科学基金青年科学基金资助项目(31501159);天津市自然科学基金资助项目(19JCZDJC36500)

摘　　要：目的观察沉默烷基甘油酮磷酸合酶(AGPS)表达对神经胶质瘤细胞长链非编码RNA(lncRNA)及其共表达mRNA表达谱的影响,并预测其生物学功能。方法将神经胶质瘤U251细胞分为shR-AGPS-1组、shR-AGPS-2组和对照组,分别转染shR-AGPS-1、shR-AGPS-2和阴性对照慢病毒,筛选单克隆细胞后观察细胞形态。采用Western blotting法检测AGPS表达,基因芯片技术检测lncRNA及其共表达mRNA的差异表达谱,对lncRNA共表达mRNA进行基因本体(GO)及京都基因与基因组大百科全书(KEGG)通路富集分析,构建信号通路作用网络、全局信号转导网络、lncRNA对通路的调控网络。结果与对照组比较,shR-AGPS-1组和shR-AGPS-2组细胞均出现团缩以及增殖抑制等变化,且shR-AGPS-2组变化更加显著。shR-AGPS-1组、shR-AGPS-2组细胞AGPS蛋白相对表达量均低于对照组,且shR-AGPS-2组降低更明显(P均<0.05)。与对照组比较,shR-AGPS-1组有73个lncRNA表达上调、107个lncRNA表达下调、13个mRNA表达上调、63个mRNA表达下调,shR-AGPS-2组266个lncRNA表达上调、233个lncRNA表达下调、61个mRNA表达上调、111个mRNA表达下调。GO富集分析显示共表达的mRNA主要参与细胞对缺氧的反应等生物学过程,KEGG富集分析显示晚期糖基化终末产物/晚期糖基化终末产物受体信号通路的富集度最高。信号通路作用网络和全局信号转导网络中连线较多的节点为磷脂酰肌醇三激酶(PI3K)信号通路、Toll样受体信号通路、血管内皮生长因子(VEGF)信号通路等。lncRNA对通路的调控网络中lncRNA AK093732处于网络调控的枢纽地位,细胞因子及其受体相互作用为差异lncRNA调控的核心通路。结论沉默AGPS表达可通过引起神经胶质瘤细胞中相关lncRNA及其共表达mRNA的差异表达而影响细胞增殖和凋亡,其中PI3K、Toll样受体及VEGF信号通路可能是其主要靶点通路。Objective To explore the effects of alkyldihydroxyacetone phosphatesyllthase(AGPS)silencing on the expression profiles of long non-coding RNAs(lncRNAs)and messenger RNAs(mRNAs),and to predict their biological functions.Methods Human glioma U251 cells were divided into the shR-AGPS-1 group,shR-AGPS-2 group,and control group,which were infected with shR-AGPS-1,shR-AGPS-2,and negative control lentivirus,then the monoclonal cells were screened and we observed their cell modality.The expression of AGPS was examined by Western blotting.The expression profiles of lncRNA and its co-expression mRNA in the glioma U251 cells were detected by microarray.The co-expression mRNAs of differential lncRNA were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways,and the signal pathway network,gene transduction network and regulation network on pathway were built.Results Compared with control group,the constriction,proliferation suppressing and decreased expression of AGPS were found in the shR-AGPS-1 and shR-AGPS-2 group,and it was more significant in the shR-AGPS-2 group(all P<0.05).The relative expression of AGPS protein in the shR-AGPS-1 group and shR-AGPS-2 group was lower than that in the control group,and the shR-AGPS-2 group had a more significant decrease(all P<0.05).Compared with control group,there were 73 up-regulated lncRNAs and 13 up-regulated mRNAs,107 down-regulated lncRNAs and 63 down-regulated mRNAs in the shR-AGPS-1 group,and there were 266 up-regulated lncRNAs and 61 up-regulated mRNAs,and 233 down-regulated lncRNAs and 111 down-regulated mRNAs in the shR-AGPS-2 group.GO analysis showed that the co-expression mRNAs were mainly involved in the biological processes such as cellular response to hypoxia.KEGG pathway analysis showed that the most enriched signal pathway was AGE-RAGE signaling pathway in diabetic complications.In the signal pathway network and gene transduction network,the signal pathways of phosphatidylinositol 3-kinase(PI3K),Toll-like receptor and vascular endotheli

关 键 词：神经胶质瘤 AGPS基因 长链非编码RNA 生物信息学 血管内皮生长因子 

分 类 号：R739.41[医药卫生—肿瘤]