circRNA_0000253 Upregulates IDD by Competitively Adsorbing miRNA-141-5p
(A) qPCR showed the miRNA-141-5p expression in the NC, circRNA_0000253 siRNA, and circRNA_0000253 siRNA + antagomiRNA-141-5p groups; ∗p < 0.05, ∗∗p < 0.01. (B) Dual-luciferase assay showed the binding effect between circRNA_0000253 and miRNA-141-5p; ∗∗p < 0.01. (C–E) The expression of circRNA_0000253 (C), miRNA-141-5p (D), and SIRT1 (E) was detected in NP samples from patients with or without IDD by RNA FISH. circRNA_0000253, miRNA-141-5p, and SIRT1 probes were labeled with Alexa 488. Nuclei were stained with DAPI. Scale bars, 100 μm. (F) WB shows the protein expression of caspase-3, caspase-7, caspase-9, MMP-3, MMP-13, ADAMTS4, ADAMTS 5, collagen II, aggrecan, and SIRT1 in the NC, circRNA_0000253 siRNA, and circRNA_0000253 siRNA + antagomiRNA-141-5p groups. (G) Flow cytometry showed the apoptosis of NPCs in the NC, circRNA_0000253 siRNA, and circRNA_0000253 siRNA + antagomiRNA-141-5p groups. (H) CCK-8 assay showed the proliferation of NPCs in the NC, circRNA_0000253 siRNA, and circRNA_0000253 siRNA+ antagomiRNA-141-5p groups.