高通量测序分析冠心病患者外周血 LncRNA表达差异
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苏州市卫生和计划生育委员会临床重点病种诊疗技术专项(LCZX201610)


Analysis the expression differences of peripheral blood LncRNA in patients with coronary artery disease by high⁃throughput sequencing
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    摘要:

    目的:发现外周血中与冠状动脉粥样硬化性心脏病(冠心病)相关的未知长链非编码RNA(LncRNA)。方法:选取临床冠心病组及对照组各3例外周血血浆样本进行LncRNA高通量测序表达谱分析,使用cuffdiff软件获得LncRNA表达谱,计算两组样本间的倍数变化和P-value值,筛选差异表达LncRNA。Q-PCR法验证36对冠心病和正常组外周血浆及外周血单核细胞中的基因表达水平。结果:测序结果分析发现差异表达明显的45个上调表达及29个下调表达的LncRNA。Q-PCR法筛选出3个在冠心病患者外周血单核细胞中上调表达明显的LncRNA:uc003pxg.1,ENST00000565257,ENST00000568324。ROC曲线下面积分别为0.812 5,0.742 4,0.742 4。以上基因在冠心病患者中上调表达均差异显著,与测序结果一致。结论:初步从冠心病患者外周血样本中筛选出上调表达的LncRNA,这为后续深入研究奠定基础,为LncRNA用于冠心病临床早期筛查提供理论依据。

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    Objective:To discover unknown LncRNAs associated with Coronary Artery Disease(CAD)in peripheral blood. Methods:Three pairs of clinical CAD group and control blood plasma samples were texted in this experiment by high-throughput sequencing methods. Cuffdiff software was used to get the expression profiles of LncRNAs,differentially expressed LncRNAs were identified based on fold change and p-value. Differently expressed genes in peripheral plasma and blood monocytes were verified by Q-PCR in 36 pairs of clinical CAD group and controls. Results:The optimal 45 up-regulated and 29 down-regulated LncRNAs were detected. Three LncRNAs uc003pxg.1,ENST00000565257,ENST00000568324 were up-regulated obviously in peripheral blood mononuclear cells of CAD patients. Area under the ROC curve results of uc003pxg.1,ENST00000565257,ENST00000568324 were 0.812 5,0.742 4,0.742 4 respectively. These genes were all up-regulated in CAD patients obviously and these outcomes were consistent with the sequencing results. Conclusion:The ideal LncRNAs were detected preliminarily. This founding lays the foundation for further research,and provides theoretical basis of LncRNA using for CAD early clinical screening.

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李 平,徐桂冬,庞 智,翁嘉懿,尹 娟,孙康云.高通量测序分析冠心病患者外周血 LncRNA表达差异[J].南京医科大学学报(自然科学版),2018,(12):1696-1700

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  • 收稿日期:2018-08-10
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  • 在线发布日期: 2019-01-02
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