小鼠胚胎成纤维细胞内质网应激反应中的长链非编码RNA表达谱及生物信息学研究
投稿时间:2014-09-22  修订日期:2014-10-18  点此下载全文
引用本文:全弘扬,樊倩倩,王林.小鼠胚胎成纤维细胞内质网应激反应中的长链非编码RNA表达谱及生物信息学研究[J].医学研究杂志,2015,44(4):61-66
DOI: 10.11969/j.issn.1673-548X.2015.04.017
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作者单位
全弘扬 100005 北京, 中国医学科学院/北京协和医学院基础医学研究所生理学系 
樊倩倩 100005 北京, 中国医学科学院/北京协和医学院基础医学研究所生理学系 
王林 100005 北京, 中国医学科学院/北京协和医学院基础医学研究所生理学系 
中文摘要:目的 探究内质网应激反应也称为未折叠蛋白效应(unfolded protein response, UPR)激活后小鼠胚胎成纤维细胞(mouse embryonic fibroblasts, MEFs)的长链非编码RNA(long noncoding RNA, lncRNA)表达谱。 方法 分离13天的小鼠胚胎获得并培养MEFs,采用内质网应激反应诱导剂衣霉素(tunicamycin)处理细胞16h后发生UPR;以未处理的MEFs作为阴性对照,采用基因芯片技术检测lncRNA表达谱,实时荧光定量PCR(real-time PCR)验证lncRNA芯片结果并进行生物信息学分析。 结果 与未使用衣霉素处理的MEFs相比, 衣霉素处理的MEFs有411个lncRNAs表达量出现显著上调,790个lncRNAs出现显著下调;实时荧光定量PCR验证部分lncRNAs表达结果与芯片一致;高级生物信息学分析分析了显著变化的lncRNAs并提示它们可能参与了细胞内许多生物学过程的调控。 结论 lncRNA 表达谱及生物信息学分析提示lncRNA在UPR激活后的细胞功能中可能发挥重要作用。
中文关键词:长链非编码RNA  内质网应激反应  基因芯片  生物信息学
 
Expression Profile and Bioinformatics Studies of Long Noncoding RNA in Mouse Embryonic Fibroblasts under Endoplasmic Reticulum Stress
Abstract:Objective To explore the potential significance of long noncoding RNA (lncRNA) expressed in endoplasmic reticulum stress response also known as unfolded protein response (UPR). Methods Mouse embryonic fibroblasts (MEFs) were isolated from 13 day mouse embryo, cultured and treated with tunicamycin for 16 hours to induce UPR. The untreated MEFs were used as negative control. The lncRNA expression profile was examined by a customized lncRNA array. The results of array assay were validated by real-time PCR and further analyzed by bioinformatics. Results There were 411 lncRNAs, whose expression was significantly up-regulated in MEFs treated with tunicamycin as compared with the controls, while 790 lncRNAs were significantly down-regulated. A number of significantly alter lncRNAs were validated by real-time PCR. A further bioinformatics analysis of lncRNA profile suggested that many could be involved in multiple cellular pathways. Conclusion Our lncRNA expression profile and bioinformatics studies strongly suggest that many lncRNAs can be regulated in the cells under UPR and could also in turn regulate UPR and other cellular processes.
keywords:Long noncoding RNA  Endoplasmic reticulum stress response  Microarray  Bioinformatics
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