Circular RNA 0047905 acts as a sponge for microRNA4516 and microRNA1227-5p, initiating gastric cancer progression

Figures & data

Figure 1. Heat map of the miRNA profiles in GC tissues versus paired normal gastric mucosa tissues.

Red indicates high relative expression, and green indicates low relative expression. The expression of miRNA was hierarchically clustered on the y-axis, and the tissue samples were hierarchically clustered on the x-axis (T vs N; Fold-change ≥ 2 or fold-change ≤ 0.5, FDR <0.05). Expression levels are presented in red and green, indicating upregulated and downregulated RNAs, respectively. Numbers marked with T and N are from five paired gastric cancer tissues and paired adjacent normal tissues, respectively.

Figure 2. circRNAs-miRNAs-mRNAs network.

A: The network consists of 591 circRNAs, 223 mRNAs and 29 miRNAs. In the circRNA-miRNA-mRNA-Net, the circle represents miRNAs, the rhombus represents mRNA and the inverted triangle represents circRNAs. Their relationship is represented by one edge. B: ceRNA network of circRNA0047905.

Figure 3. The Kaplan-Meier survival analysis of SERPINB5 and MMP11 in GC patients.

A: The Kaplan-Meier survival analysis and log-rank test showing up-regulation of SERPINB5 correlates with low overall survival rates (HR = 1.92, *P < 0.05, n = 93). B: The Kaplan-Meier survival analysis and log-rank test show up-regulation of MMP11 correlates with low overall survival rates (HR = 1.61, *P < 0.05, n = 876).

Figure 4. Relative expression of MMP11 in human GC cell lines transfected with specific siRNA targeted circRNA0047905.

A: qRT-PCR on MMP11 expression in 31 paired GC tissues and adjacent non-cancer tissues. B: Relative expression of MMP11 mRNAs in AGS GC cell lines transfected with specific siRNAs targeted circRNA0047905, *P < 0.05; C: Expression of MMP11 protein in AGS GC cell lines transfected with specific siRNAs targeted circRNA0047905.

Figure 5. Overexpression of circRNA0047905 increased luciferase activity of SERPINB5-3ʹUTR-WT reporter by binding endogenous miR4516.

A: Schematic of circRNA0047905 wild-type (WT) and mutant (Mut) luciferase reporter vectors. B: The relative luciferase activities were analyzed in 293T cells co-transfected with miR4516 mimics or miR-NC and luciferase reporter vectors circRNA0047905-WT or circRNA0047905-Mut. C: Schematic of SERPINB5 wild-type (WT) and mutant (Mut) luciferase reporter vectors. D: The relative luciferase activities were analyzed in 293T cells co-transfected with miR4516 mimics or miR-NC and luciferase reporter vectors SERPINB5-3ʹUTR-WT or SERPINB5-3ʹUTR-Mut. E: The relative luciferase activities were analyzed in 293T cells co-transfected with miR4516 mimics or miR-NC, pcDNA3.1-circRNA0047905 overexpression vector or pcDNA3.1-control and luciferase reporter vectors SERPINB5-3ʹUTR-WT.

Figure 6. Overexpression of circRNA0047905 increased luciferase activity of MMP11-3ʹUTR-WT reporter by binding endogenous miR1227-5p.

A: Schematic of circRNA0047905 wild-type (WT) and mutant (Mut) luciferase reporter vectors. B: The relative luciferase activities were analyzed in 293T cells co-transfected with miR1227-5p mimics or miR-NC and luciferase reporter vectors circRNA0047905-WT or circRNA0047905-Mut. C: Schematic of MMP11 wild-type (WT) and mutant (Mut) luciferase reporter vectors. D: The relative luciferase activities were analyzed in 293T cells co-transfected with miR1227-5p mimics or miR-NC and luciferase reporter vectors MMP11-3ʹUTR-WT or MMP11-3ʹUTR-Mut. E: The relative luciferase activities were analyzed in 293T cells co-transfected with miR1227-5p mimics or miR-NC, pcDNA3.1-circRNA0047905 overexpression vector or pcDNA3.1-control and luciferase reporter vectors MMP11-3ʹUTR-WT.

Figure 7. Down-regulated circRNA0047905 expression in gastric cancer cells inhibits Akt/CREB signaling pathway activation.

Figure 8. RNA scope assay targeting circRNA0047905 evaluation in histological samples.

N: adjacent noncancerous mucosal tissue samples; T: gastric carcinoma samples (× 40). Positive signals of probe were indicated by red dots in GC tissues.

Figure 9. The schematic diagram shows the mechanism underlying circRNA0047905 as a ceRNA for miR4516 and miR1227-5p.