Research Paper

Figure 6. LINC01224 regulates the expression of YES1, a target gene of miR-193a-5p. (A) Prediction of potential genes competing with LINC01224 for adsorption of miR-193a-5p. We first use the online websites (miRDB, mirDIP and miRWalk) to predict the target genes of miR-193a-5p, and then intersect with the co-expressed genes of LINC01224 to obtain the potential target genes of LINC01224. (B) The effect of miR-193a-5p mimics on the expression of potential target genes was detected by qRT-PCR. (C) Western blot analysis of YES1 expression in AGS cells by overexpressing miR-193a-5p. (D) Dual-luciferase reporter assay showed that miR-193a-5p mimics significantly decreased the luciferase activity containing wild-type 3'-UTR segment of YES1 plasmid rather than mutant-3'-UTR segment of YES1 plasmid. (E) Dual-luciferase reporter assay showed that overexpression of wt-LINC01224 could partially eliminate the inhibitory effects of miR-193a-5p mimics on the luciferase activity of plasmid containing wild-type 3'-UTR segment of YES1. (F) Western blot showed that miR-193a-5p mimics decreased YES1 expression, while overexpression of LINC01224 partially reversed the inhibition of YES1 expression by miR-193a-5p mimics. (G) Western blot showed that wild-type LINC01224 enhanced YES1 expression, whereas the mutated LINC01224 containing a mutation in the binding site of miR-193a-5p could not increase YES1 expression. (H) Immunohistochemical staining showed that YES1 was highly expressed in gastric cancer compared with paracancerous tissues. Scale bar = 50 μm. (I) Immunohistochemical staining showed that YES1 expression was significantly decreased in xenografts with LINC01224 knockdown compared with control tissues. Scale bar = 50 μm.