Research Paper Volume 11, Issue 23 pp 11202—11223

ciRs-6 upregulates March1 to suppress bladder cancer growth by sponging miR-653

Figure 5. ciRs-6 sponges miR-653 to elevate March1 levels. (A) Target genes that binds to miR-653 were predicted by TargetScan, RNAhybrid, and miRDB; (B) qPCR was used to detect the expression of the 17 predicted targets following ciRs-6 overexpression; (C) miR-653 significantly reduced March1 expression in bladder cancer cells; (D, E) dual luciferase reporter assays showed binding between March1 and miR-653. (F, G) qPCR and Western blot experiments were performed to detect March1 levels among ciRs-6+miR-nc, vector+miR-nc, ciRs-6+miR-653, and vector+miR-653 cells. (H) IHC was performed to detect March1 levels in tissues that were previously collected from a subcutaneous tumor model. Pictures were photographed at 200× under a light microscope. The results are displayed as the mean±SEM, *p<0.05, **p<0.01.