Fig 1.
Differential expression of circRNAs between human acute Stanford type A aortic dissection (AAAD) tissues and normal aortic tissues.
(A) The flowchart of this study. (B) Hematoxylin and eosin (HE)–stained sections showing the AAAD. (C) Hierarchical cluster analysis of all target circRNAs. 506 circRNAs were significantly differentially expressed (P<0.05, false discovery rate, FDR<0.05, fold change>2) between human AAAD tissues and normal aortic tissues. Among these, 320 circRNAs were upregulated and 186 circRNAs were downregulated in AAAD tissues. (D) Differentially expressed circRNAs were displayed by volcano plots. The blue and red parts indicated >2 fold decreased and -increased expression of the dysregulated circRNAs in AAAD tissues, respectively (P< 0.05). (E) Classification of differentially expressed circRNAs.
Table 1.
Clinical characteristics of patients who were enrolled in this study.
Fig 2.
qRT-PCR analysis of differentially expressed circRNAs, miRs and mRNAs.
The relative expression levels of the selected significantly differentially expressed circRNAs (A-J), miRs (K-O) and mRNAs (P-T) in 30 matched human AAAD and normal aortic tissues. The value of 2-ΔΔCt was used to show the expression level of RNAs. Data are expressed as mean±SD. ****P < 0.0001.
Fig 3.
GO, KEGG pathway enrichment analysis and the gene set enrichment analysis (GSEA) of the target genes of differentially expressed circRNAs.
(A) Biological process of GO analysis; (B) Cellular component of GO analysis; (C) Molecular function of GO analysis; (D) KEGG pathways analysis; (E) Scatterplot of enriched KEGG pathways. Y-axis represents pathway name and X-axis represents rich factor. Size and color of each bubble represents the number of differentially expressed genes enriched in the pathway and -log10(q-value), respectively. (F-K) GSEA enrichment plot. The GSEA enrichment plot for cell cycle (F), cell death (G), cellular component disassembly (H), response to DNA damage stimulus (I), regulation of canonical Wnt signaling pathway (J) and regulation of response to stress (K).
Fig 4.
Weighted gene correlation network analysis (WGCNA) of the target genes of differentially expressed circRNAs.
(A) The hierarchical cluster dendrogram of the target genes of differentially expressed circRNAs and the color assignments for each module. (B) The module-trait (sample) correlations and corresponding P-values. (C) The module-group correlations and corresponding P-values. (D) Gene co-expression network of the genes enriched in grey module.
Fig 5.
Prediction of circMARK3-miR-1273g-3p-Fgr interaction.
(A) miR-1273g-3p targeting Fgr. (B) miR-1273g-3p targeting circMARK3. (C) Prediction of circMARK3-miRNA-mRNA interaction. The base-pairing and the minimum free energy (mfe) were predicted using the RNA hybrid program. (D-F) The relative expression levels of Fgr in the tissue of AAAD patients measured by qRT-PCR (D) and western blotting (E-F). (G-H) The relative expressions of miR-1273g-3p and circMARK3 in the tissue of AAAD patients. (I-L) HASMCs were transfected with circMARK3 or control vector. The relative expression of circMARK3 and hsa-miR-1273g-3p in HASMCs were measured by qRT-PCR assays (I-L). The expression of Fgr in the HASMCs were measured by western blotting assays (K-L). The value of 2-ΔΔCt was used to show the expression level of RNAs. Data are expressed as mean±SD. **P < 0.01, ***P < 0.001 and ****P < 0.0001.
Fig 6.
The diagnostic value analysis of serum circMARK3 for AAAD diagnosis.
(A) circMARK3 was up-regulated in the serum of AAAD group. The value of 2-ΔΔCt was used to show the expression level of RNAs. Data are expressed as mean±SD. ****P < 0.0001. (B) Receiver operating characteristic (ROC) curve showed the diagnostic value of serum circMARK3 for AAAD diagnosis (AUC = 0.9344, 95% CI: 0.8677–1.001, P< 0.0001).