Tian Jiangtian, Fu Yahong, Li Qi, Xu Ying, Xi Xiangwen, Zheng Yuqi, Yu Li, Wang Zhuozhong, Yu Bo, Tian Jinwei
Key Laboratory of Myocardial Ischemia, Chinese Ministry of Education, Harbin, China.
Department of Cardiology, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Front Genet. 2020 May 29;11:530. doi: 10.3389/fgene.2020.00530. eCollection 2020.
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.
动脉粥样硬化由多种因素介导,是心脑血管疾病重要的病理基础。血管平滑肌细胞(VSMC)异常增殖和迁移在动脉粥样硬化病变形成中起关键作用。环状RNA(circRNA)已在不同物种中被广泛检测到,且与多种疾病密切相关。然而,circRNA在VSMC中的表达谱和分子调控机制仍不清楚。我们使用高通量RNA测序以及生物信息学工具,系统分析了来自不同VSMC表型样本中的circRNA表达谱。通过聚合酶链反应(PCR)、桑格测序和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,共检测到对应于6273个基因(宿主基因)的22191条circRNA,在PDGF-BB处理组和对照组之间鉴定出了112条差异表达的circRNA,其中59条上调,53条下调。我们选择了9条circRNA评估其特定的头对尾剪接,并选择两组之间10条差异表达的circRNA进行qRT-PCR验证。基因本体论和京都基因与基因组百科全书分析富集分析表明,circRNA的亲本基因主要参与心肌原纤维组装和DNA模板转录的正调控,表明它们可能参与心血管疾病。最后,我们基于失调的circRNA和VSMC相关的微小RNA构建了一个circRNA-微小RNA网络。我们的研究首次展示了circRNA在PDGF-BB诱导的VSMC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。