Figure 4.

CASC9 functions as a sponge for miR-758-3p. (A) The relative expression levels of miR-758-3p in BC cell lines (TCCSUP, UM-UC-3, T24, J82 and 5637) compared with normal human bladder epithelial cells (SV-HUC-1). (B) CASC9 functions as a sponge for miR-758-3p. Upper panel, schematic diagrams of the mutual interactions between miR-758-3p and CASC9. Bottom panel, 293T cells were transfected with miRNA mimics in combination with luciferase reporters harboring wild-type or mutated miRNA binding sites on CASC9. The effects of miR-758-3p on luciferase activity were determined by luciferase reporter assays. The activities of firefly luciferase were normalized to Renilla luciferase. *P<0.05. (C) Transwell assay of 5637 and J82 cells transfected with mimics NC, mimics miR-758-3p, inhibitor NC or inhibitor miR-758-3p as indicated reported as the means ± SD from 3 independently repeated experiments. (D) Wound healing assays of 5637 and J82 cells transfected with NC mimics, miR-758-3p mimics, NC inhibitor or miR-758-3p inhibitor as indicated. Unpaired Student's t-test was used and data are presented as the mean ± SEM. Scale bars, 100 µm. *P<0.05, **P<0.01 and ***P<0.001. CASC9, cancer susceptibility candidate 9; miR-758-3p, microRNA-758-3p; BC, bladder cancer; NC, negative control; SEM, standard error of mean.