Differential Expression and Bioinformatics Analysis of CircRNA in PDGF-BB-Induced Vascular Smooth Muscle Cells
Differential Expression and Bioinformatics Analysis of CircRNA in PDGF-BB-Induced Vascular Smooth Muscle Cells
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DOI：
10.3389/fgene.2020.00530
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发表时间：
2020-05-29
影响因子：
3.7
通讯作者：
Tian, Jinwei
中科院分区：
文献类型：
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作者：
Tian, Jiangtian;Fu, Yahong;Tian, Jinwei
动脉粥样硬化是多种因素介导的，是心脑血管疾病的重要病理基础。异常血管平滑肌细胞（VSMCs）的增殖和迁移在动脉粥样硬化病变的形成中起重要作用。环状rna （circRNA）广泛存在于不同物种中，与多种疾病密切相关。然而，环状rna在VSMCs中的表达谱和分子调控机制尚不清楚。我们使用高通量RNA-seq和生物信息学工具系统地分析了不同VSMC表型样品中的circRNA表达谱。采用聚合酶链反应（PCR）、Sanger测序和qRT-PCR对circRNA进行验证。在血小板衍生生长因子（PDGF-BB）处理组、空白对照组或两组中共检测到22191个circrna，对应6273个基因（宿主基因），在PDGF-BB处理组和对照组之间共鉴定出112个差异表达的circrna，其中59个表达上调，53个表达下调。我们选择了9个circrna来评估特异性头尾剪接，并在两组之间选择了10个差异表达的circrna进行qRT-PCR验证。基因本体和京都基因与基因组百科分析富集分析显示，circRNAs亲本基因主要参与心肌原纤维组装和dna模板转录的正向调控，提示其可能参与心血管疾病。最后，我们基于失调的circrna和vsmc相关的microrna构建了一个circRNA-miRNA网络。我们的研究首次发现了环状rna在pdgf - bb诱导的VSMCs中的差异表达，可能为血管疾病的预防和治疗提供新的思路和靶点。
Atherosclerosis is mediated by various factors and plays an important pathological foundation for cardiovascular and cerebrovascular diseases. Abnormal vascular smooth muscle cells (VSMCs) proliferation and migration have an essential role in atherosclerotic lesion formation. Circular RNAs (circRNA) have been widely detected in different species and are closely related to various diseases. However, the expression profiles and molecular regulatory mechanisms of circRNAs in VSMCs are still unknown. We used high-throughput RNA-seq as well as bioinformatics tools to systematically analyze circRNA expression profiles in samples from different VSMC phenotypes. Polymerase chain reaction (PCR), Sanger sequencing, and qRT-PCR were performed for circRNA validation. A total of 22191 circRNAs corresponding to 6273 genes (host genes) in the platelet-derived growth factor (PDGF-BB) treated group, the blank control group or both groups, were detected, and 112 differentially expressed circRNAs were identified between the PDGF-BB treated and control groups, of which 59 were upregulated, and 53 were downregulated. We selected 9 circRNAs for evaluation of specific head-to-tail splicing, and 10 differentially expressed circRNAs between the two groups for qRT-PCR validation. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses enrichment analyses revealed that the parental genes of the circRNAs mainly participated in cardiac myofibril assembly and positive regulation of DNA-templated transcription, indicating that they might be involved in cardiovascular diseases. Finally, we constructed a circRNA-miRNA network based on the dysregulated circRNAs and VSMC-related microRNAs. Our study is the first to show the differential expression of circRNAs in PDGF-BB-induced VSMCs and may provide new ideas and targets for the prevention and therapy of vascular diseases.