Figures & data
Figure 1. Lnc-DGCR5 is up-regulated in ASMCs treated by PDGF-BB. (a) The expression level of lnc-DGCR5 increased in ASMCs treated with PGE2, ET-1, TGF-β1, and PDGF-BB. (b) The expression level of lnc-DGCR5 increased in PDGF-BB-treated ASMCs in a time-dependent manner. *p < 0.05 vs vehicle or 0 groups, **p < 0.01 vs vehicle or 0 groups.
Figure 2. Lnc-DCR5 promotes the proliferation and migration of ASMCs. (a) Inhibiting the expression of lnc-DGCR5 was obtained through the siRNAs. (b) CCK-8 assay indicated that the cell viability of ASMCs treated by PDGF-BB is weakened when the expression of lnc-DGCR5 was inhibited. (c) Colony formation assay showed the significantly decreased proliferation ability of ASMCs when the expression of lnc-DGCR5 was inhibited. (d) Migrated ability of ASMCs, as observed using the Transwell assay, showed the migration of ASMCs after transfection. (e) Western blotting confirmed the effect of lnc-DGCR5 on PCNA, MMP-3, and MMP-9 expression in PDGF-BB-treated ASMCs. *p < 0.05, **p < 0.01.
Figure 3. miR-204-5p acts as the target of lnc-DGCR5 in the ASMCs. (a) Bioinformatic analysis for the binding sites between miR-210-5p and lnc-DGCR5. (b) Luciferase assay was used to detect the direct interaction between miR-204-5p and lnc-DGCR5. (c) RNA pull-down assay confirmed that miR-204-5p can directly target lnc-DGCR5. (d) The expression level of miR-204-5p in the ASMCs was downregulated when lnc-DGCR5 was overexpressed, and its expression level was increased when lnc-DGCR5 was inhibited. (e) The expression level of miR-204-5p was downregulated in ASMCs treated by PDGF-BB, and knockdown of lnc-DGCR5 reversed the downregulation. **p < 0.01.
Figure 4. MiR-204-5p inhibition reversed the effect of lnc-DGCR5 knockdown. (a) The inhibitor can effectively inhibit the expression of miR-204-5p. (b) CCK-8 analysis showed that miR-204-5p inhibitor can rescue the cell viability in PDGF-BB-treated AMSC when lnc-DGCR5 was inhibited. (c) The results of the colony formation assay showed the miR-204-5p inhibitor can rescue proliferation ability in PDGF-BB-treated AMSC when lnc-DGCR5 was inhibited. (d) The results of the Transwell assay showed that the miR-204-5p inhibitor can rescue the migration ability of PDGF-BB-treated AMSCs when lnc-DGCR5 was inhibited. (e) Western blotting results showed that miR-204-5p inhibitor can rescue the expression level of PCNA, MMP-9, and MMP-3 in PDGF-BB-treated AMSCs when lnc-DGCR5 was inhibited. *p < 0.05, **p < 0.01.
Figure 5. miR-204-5p targets SRSF7 directly. (a) The enrichment analysis of miR-204-5p targets-related biological pathway. (b) The binding site of miR-204-5p in SRSF7 3’-UTR. (c) The luciferase assay showed that miR-204-5p could directly targets SRSF7. (d) RNA pull-down assay confirmed that miR-204-5p can directly target SRSF7. (e) The expression level of SRSF7 in the ASMCs was downregulated when miR-204-5p was overexpressed, and its expression level increased when miR-204-5p was inhibited. (f) knockdown of lnc-DGCR5 decreased SRSF7 expression, and inhibition of miR-204-5p reversed the decrease. (g) The expression level of SRSF7 was upregulated in ASMCs treated by PDGF-BB. (h) Protein level of SRSF7 increased in ASMCs treated with PDGF-BB. **p < 0.01.
Figure 6. Overexpression of SRSF7 can rescue the proliferation and migration of ASMC treated by PDGF-BB, which is attenuated by overexpression of miR-204-5p. (a) qRT-PCR confirmed the overexpression efficiency of SRSF7. (b) CCK-8 analysis showed that overexpression of SRSF7 can rescue the cell viability in PDGF-BB-treated AMSC when miR-204-5p was overexpressed. (c) The results of the colony formation assay showed that overexpression of SRSF7 can rescue the proliferation ability in PDGF-BB-treated AMSC when miR-204-5p was overexpressed. (d) The results of Transwell assay showed that overexpression of SRSF7 can rescue migration ability in PDGF-BB-treated AMSC when miR-204-5p was overexpressed. (e) Western blotting showed that overexpression of SRSF7 can rescue the expression level of PCNA, MMP-9, and MMP-3 in PDGF-BB-treated AMSC when miR-204-5p was overexpressed. *p < 0.05, **p < 0.01.
Figure S1. Knockdown of Lnc-DGCR5 has no significant difference on the proliferation and migration of ASMCs. (a) The cell viability of AMSCs. (b) Colony formation of AMSCs. (c) The invasion of AMSCs. (d) Protein expression of PCNA, MMP-3, and MMP-9 in AMSCs.
