Peng Wenjuan, Cao Han, Liu Kuo, Guo Chunyue, Sun Yanyan, Qi Han, Liu Zheng, Xie Yunyi, Liu Xiaohui, Li Bingxiao, Zhang Ling
Department of Epidemiology and Health Statistics, School of Public Health, Capital Medical University, Beijing Municipal Key Laboratory of Clinical Epidemiology Beijing 100069, People's Republic of China.
The National Clinical Research Center for Mental Disorders and Beijing Key Laboratory of Mental Disorders and The Advanced Innovation Center for Human Brain Protection, Beijing Anding Hospital, School of Mental Health, Capital Medical University Beijing 100088, People's Republic of China.
Am J Transl Res. 2020 Oct 15;12(10):6060-6075. eCollection 2020.
To identify long noncoding RNAs (lncRNAs) and construct a competing endogenous RNA (ceRNA) network for essential hypertension.
An RNA microarray and two-step quantitative real-time PCR were applied to identify differentially expressed RNAs (DE-RNAs), and a luciferase assay was performed to explore the binding relationship between RNAs. A generalized linear model and logistic regression model were used to analyze the associations between different RNAs and of RNAs with hypertension. Receiver operating characteristic curve analysis was executed to evaluate the diagnostic performance. Bioinformatics analysis was applied for network construction.
In total, 439 DE-RNAs (387 lncRNAs and 52 mRNAs) were identified in the microarray, and 71 'lncRNA-miRNA-mRNA' loops formed the ceRNA network. The first validation confirmed that five RNAs (NR_104160, lnc-GPR63-8:1, lnc-HPRT1-9:1, and ) were significantly upregulated in hypertensives ( < 0.05). NR_104160 was significantly associated with hypertension (OR = 2.863, 95% CI: 1.143-7.172; = 0.025) after adjusting for confounding factors. NR_104160 was included in the hypertension diagnostic model, with an area under the curve of 0.852 (95% CI: 0.761-0.944). In the second validation, NR_104160 showed a constant significant difference ( = 0.001). An elevated expression level of NR_104160 was associated with the expression of ( = 0.2235, = 0.005). Luciferase assays showed hsa-miR-101-3p stimulation significantly inhibited the reporter gene activation ability of the NR_104160 wild-type plasmid ( < 0.001).
Our study constructed a ceRNA network to provide hypotheses regarding the mechanism of hypertension development. lncRNA-NR_104160 was identified as a hub element that participates in hypertension transcriptional regulation and as a potential biomarker.
鉴定长链非编码RNA（lncRNA）并构建原发性高血压的竞争性内源性RNA（ceRNA）网络。
应用RNA微阵列和两步定量实时PCR鉴定差异表达RNA（DE-RNA），并进行荧光素酶测定以探究RNA之间的结合关系。使用广义线性模型和逻辑回归模型分析不同RNA之间以及RNA与高血压之间的关联。进行受试者工作特征曲线分析以评估诊断性能。应用生物信息学分析进行网络构建。
在微阵列中总共鉴定出439个DE-RNA（387个lncRNA和52个mRNA），71个“lncRNA-miRNA-mRNA”环形成了ceRNA网络。首次验证证实，5种RNA（NR_104160、lnc-GPR63-8:1、lnc-HPRT1-9:1和 ）在高血压患者中显著上调（<0.05）。校正混杂因素后，NR_104160与高血压显著相关（OR = 2.863，95% CI：1.143 - 7.172； = 0.025）。NR_104160被纳入高血压诊断模型，曲线下面积为0.852（95% CI：0.761 - 0.944）。在第二次验证中，NR_104160显示出持续的显著差异（ = 0.001）。NR_104160表达水平升高与 的表达相关（ = 0.2235， = 0.005）。荧光素酶测定显示，hsa-miR-101-3p刺激显著抑制了NR_104160野生型质粒的报告基因激活能力（<0.001）。
我们的研究构建了一个ceRNA网络，为高血压发病机制提供了假设。lncRNA-NR_104160被鉴定为参与高血压转录调控的枢纽元件和潜在生物标志物。
