College of Life Science, Zhejiang Chinese Medical University, Hangzhou, 310053, Zhejiang Province, China.
Zhejiang University School of Medicine, Hangzhou, 310058, Zhejiang Province, China.
Mol Cell Biochem. 2018 Feb;439(1-2):171-187. doi: 10.1007/s11010-017-3146-2. Epub 2017 Aug 18.
This study aimed to systematically investigate the relationship between miRNA expression and the occurrence of ventricular septal defect (VSD), and characterize the miRNA target genes and pathways that can lead to VSD. The miRNAs that were differentially expressed in blood samples from VSD and normal infants were screened and validated by implementing miRNA microarrays and qRT-PCR. The target genes regulated by differentially expressed miRNAs were predicted using three target gene databases. The functions and signaling pathways of the target genes were enriched using the GO database and KEGG database, respectively. The transcription and protein expression of specific target genes in critical pathways were compared in the VSD and normal control groups using qRT-PCR and western blotting, respectively. Compared with the normal control group, the VSD group had 22 differentially expressed miRNAs; 19 were downregulated and three were upregulated. The 10,677 predicted target genes participated in many biological functions related to cardiac development and morphogenesis. Four target genes (mGLUR, Gq, PLC, and PKC) were involved in the PKC pathway and four (ECM, FAK, PI3 K, and PDK1) were involved in the PI3 K-Akt pathway. The transcription and protein expression of these eight target genes were significantly upregulated in the VSD group. The 22 miRNAs that were dysregulated in the VSD group were mainly downregulated, which may result in the dysregulation of several key genes and biological functions related to cardiac development. These effects could also be exerted via the upregulation of eight specific target genes, the subsequent over-activation of the PKC and PI3 K-Akt pathways, and the eventual abnormal cardiac development and VSD.
本研究旨在系统研究 miRNA 表达与室间隔缺损(VSD)发生之间的关系,并阐明导致 VSD 的 miRNA 靶基因和途径。通过实施 miRNA 微阵列和 qRT-PCR 筛选和验证了在 VSD 和正常婴儿血液样本中差异表达的 miRNAs。使用三个靶基因数据库预测差异表达 miRNA 调控的靶基因。使用 GO 数据库和 KEGG 数据库分别对靶基因的功能和信号通路进行富集。使用 qRT-PCR 和 Western blot 分别比较 VSD 组和正常对照组中关键途径中特定靶基因的转录和蛋白表达。与正常对照组相比,VSD 组有 22 个差异表达的 miRNAs;19 个下调,3 个上调。10677 个预测的靶基因参与了许多与心脏发育和形态发生相关的生物学功能。四个靶基因(mGLUR、Gq、PLC 和 PKC)参与了 PKC 途径,四个靶基因(ECM、FAK、PI3K 和 PDK1)参与了 PI3K-Akt 途径。这八个靶基因在 VSD 组中的转录和蛋白表达显著上调。VSD 组中失调的 22 个 miRNAs 主要下调,这可能导致与心脏发育相关的几个关键基因和生物学功能失调。这些影响也可以通过八个特定靶基因的上调、随后 PKC 和 PI3K-Akt 途径的过度激活以及最终的异常心脏发育和 VSD 来发挥作用。