Department of Neurology, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Department of Internal Medicine, The Second Affiliated Hospital of Guangxi Medical University, Nanning, China.
Sci Rep. 2020 Jun 23;10(1):10138. doi: 10.1038/s41598-020-66990-y.
Recent studies have found that circular RNAs (circRNAs) play crucial roles not only in the normal growth and the development of different tissues and organs but also in the pathogenesis and progression of various disorders. However, the expression patterns and the function of circRNAs in acute ischemic stroke (AIS) in the South Chinese Han population are unclear. In the present study, RNA sequencing (RNA-seq) data was generated from 3 AIS patients and 3 healthy controls. The circRNAs were detected and identified by CIRI2 and Find_circ software. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses were used to detect the expression of circRNAs. Meanwhile, the potential diagnostic value of the selected circRNAs for AIS was assessed by generating receiver operating characteristic (ROC) curve with area under curve (AUC). The bioinformatic analysis of the host genes of differentially expressed (DE) circRNAs was performed by gene ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, KOBAS for pathway analysis and regulatory network analysis. miRNA-circRNA and miRNA-mRNA interactions were predicted by using TargetScan, miRanda and starBase. CircRNA-miRNA-mRNA interaction networks were created with Cytoscape. Our result showed that there were 2270 DE circRNAs between AIS patients and healthy controls. Among them, 659 were found upregulated and 1611 were downregulated. Bioinformatic analysis showed that the DE circRNAs were related to the following biological processes: endocytosis, energy metabolism, apoptosis, FoxO signaling pathway, platelet activation, neurotrophin signaling pathway and VEGF signaling pathway, which may be associated with the pathological of AIS. Three randomly selected circRNAs were successfully validated by qRT-PCR. The results show that hsa_circ_0005548 was significantly upregulated, while hsa_circ_0000607 and hsa_circ_0002465 were significantly downregulated in AIS. Furthermore, the AUC values for hsa_circ_005548, hsa_circ_0000607 and hsa_circ_0002465 were 0.51, 0.75 and 0.69, respectively, suggesting that hsa_circ_0000607 and hsa_circ_0002465 could be potential biomarkers for AIS. In addition, Bcl2 was predicted to be a direct target of miR-337-3p, and hsa_circRNA_0000607 was predicted to act as a sponge for miR-337-3p. Thus, hsa_circ_0000607 may be involved in AIS by regulating the miR-337-3p/Bcl2 axis. Collectively, our findings indicate that numerous dysregulated circRNAs may play pivotal functional roles in AIS and hsa_circ_0000607 may play a crucial role in the pathogenesis and progression of AIS by regulating the miR-337-3p/Bcl2 axis.
最近的研究发现,环状 RNA(circRNA)不仅在不同组织和器官的正常生长和发育中起着至关重要的作用,而且在各种疾病的发病机制和进展中也起着至关重要的作用。然而,circRNA 在华南汉族人群急性缺血性脑卒中(AIS)中的表达模式和功能尚不清楚。本研究对 3 名 AIS 患者和 3 名健康对照者的 RNA 测序(RNA-seq)数据进行了分析。利用 CIRI2 和 Find_circ 软件检测和鉴定 circRNA。采用定量逆转录-聚合酶链反应(qRT-PCR)分析检测 circRNA 的表达。同时,通过生成曲线下面积(AUC)的接收器操作特性(ROC)曲线来评估所选 circRNA 对 AIS 的潜在诊断价值。通过基因本体(GO)富集、京都基因与基因组百科全书(KEGG)通路分析、KOBAS 通路分析和调控网络分析,对差异表达(DE)circRNA 的宿主基因进行了生物信息学分析。利用 TargetScan、miRanda 和 starBase 预测了 miRNA-circRNA 和 miRNA-mRNA 相互作用。利用 Cytoscape 构建了 circRNA-miRNA-mRNA 相互作用网络。我们的结果表明,AIS 患者与健康对照组之间存在 2270 个 DE circRNA。其中 659 个上调,1611 个下调。生物信息学分析表明,DE circRNA 与以下生物学过程有关:内吞作用、能量代谢、细胞凋亡、FoxO 信号通路、血小板激活、神经营养因子信号通路和 VEGF 信号通路,这可能与 AIS 的发病机制有关。通过 qRT-PCR 成功验证了 3 个随机选择的 circRNA。结果表明,hsa_circ_0005548 明显上调,而 hsa_circ_0000607 和 hsa_circ_0002465 在 AIS 中明显下调。此外,hsa_circ_005548、hsa_circ_0000607 和 hsa_circ_0002465 的 AUC 值分别为 0.51、0.75 和 0.69,表明 hsa_circ_0000607 和 hsa_circ_0002465 可能是 AIS 的潜在生物标志物。此外,预测 Bcl2 是 miR-337-3p 的直接靶标,hsa_circRNA_0000607 被预测为 miR-337-3p 的海绵。因此,hsa_circ_0000607 可能通过调节 miR-337-3p/Bcl2 轴参与 AIS。总之,我们的研究结果表明,大量失调的 circRNA 可能在 AIS 中发挥重要的功能作用,hsa_circ_0000607 可能通过调节 miR-337-3p/Bcl2 轴在 AIS 的发病机制和进展中发挥关键作用。