Fig. 6. miR-513a-5p promotes the progression of ESCC via targeting p53 and RBM25 mediates p53 regulation of cCNTNAP3 biogenesis.

a The mRNA level of p53 in Eca-109 and 293T cells after knockdown or overexpression of cCNTNAP3 was determined by qRT-PCR. b Upper panel: miR-513a-5p can bind to the 3′-UTR of p53 mRNA; lower panel: Schematic graph illustrated the mutation of potential binding site between miR-513a-5p and the 3′-UTR regions of p53. c The direct binding between p53 3′-UTR and miR-513a-5p was analyzed by dual-luciferase reporter assay. d, e The expression levels of p53 protein and its downstream p21, cyclin D1, CDK-4, cyclin E1, CDK-2, p-Rb, E2F1, and Caspase-3 in Eca-109 and 293T cells with knockdown or overexpression of cCNTNAP3 were detected by western blot. f, g p53 levels were detected by western blot after cCNTNAP3 knockdown or overexpression and cotransfection with miR-513a-5p inhibitor or mimic. h qRT-PCR detected cCNTNAP3 expression level in Eca-109 cells transfected with p53-specific siRNA (si-p53) or p53-overexpression plasmid (OE-p53). i Expression level of RBM25 was determined by qRT-PCR in Eca-109 cells after knockdown or overexpression of p53. j Expression of cCNTNAP3 was determined by qRT-PCR in Eca-109 cells transfected with RBM25-specific siRNA (si-RBM25) or RBM25-overexpression plasmid (OE-RBM25). k qRT-PCR detected the expression of cCNTNAP3 in Eca-109 cells transfected with OE-RBM25 alone or together with si-p53. l cCNTNAP3 and RBM25 mRNA levels were measured by qRT-PCR in ESCC tissues. All the results were shown as mean ± SD (n = 3), which were three separate experiments performed in triplicate. *p < 0.05, **p < 0.01, ***p < 0.001 (Student’s t test).