Jing Ling, Jin Chengmei, Lu Ying, Huo Pingyan, Zhou Lijun, Wang Ye, Tian Ye
Department of Cardiology, First Clinical College of Harbin Medical University, Harbin, PR China.
Cardiology. 2015;130(4):223-33. doi: 10.1159/000370028. Epub 2015 Mar 18.
We aimed to investigate the differentially expressed microRNAs (miRNAs) and their target genes in human alcoholic cardiomyopathy (ACM).
The expression levels of plasma miRNAs of 78 male ACM patients and 78 healthy men were detected by using the 6th-generation miRCURY™ LNA array (v.16.0). The prediction analysis for microarrays (PAM) method was used to identify the differentially expressed miRNAs. Target genes of the identified differentially expressed miRNAs were predicted using TargetScan 5.2 and Miranda. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to perform functional annotation and pathway enrichment analysis of target genes respectively, followed by real-time RT-PCR analysis to validate the expression changes of miRNAs.
Twenty-one differentially expressed miRNAs were identified. Nine differentially expressed miRNAs (hsa-miR-506, hsa-miR-1285, hsa-miR-512-3P, hsa-miR-138, hsa-miR-485-5P, hsa-miR-4262, hsa-miR-548c-3P, has-miR-548a-5P and kshv-miR-K12-1), involved in multiple functions and pathways, were selected for real-time RT-PCR confirmation. Moreover, two significantly important subpathways (neurotrophin signaling pathway and inositol phosphate metabolism) were predicted.
The screened differentially expressed miRNAs may be involved in the development of ACM. Specific miRNAs, such as miR-138, may be considered as a new target for the early diagnosis and treatment of human ACM.
我们旨在研究人类酒精性心肌病(ACM)中差异表达的微小RNA(miRNA)及其靶基因。
使用第6代miRCURY™ LNA芯片(v.16.0)检测78例男性ACM患者和78例健康男性血浆miRNA的表达水平。采用微阵列预测分析(PAM)方法鉴定差异表达的miRNA。使用TargetScan 5.2和Miranda预测已鉴定的差异表达miRNA的靶基因。分别使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)对靶基因进行功能注释和通路富集分析,随后进行实时RT-PCR分析以验证miRNA的表达变化。
鉴定出21种差异表达的miRNA。选择9种差异表达的miRNA(hsa-miR-506、hsa-miR-1285、hsa-miR-512-3P、hsa-miR-138、hsa-miR-485-5P、hsa-miR-4262、hsa-miR-548c-3P、has-miR-548a-5P和kshv-miR-K12-1)进行实时RT-PCR确认,这些miRNA涉及多种功能和通路。此外,预测了两个重要的子通路(神经营养因子信号通路和肌醇磷酸代谢)。
筛选出的差异表达miRNA可能参与ACM的发生发展。特定的miRNA,如miR-138,可被视为人类ACM早期诊断和治疗的新靶点。