Figure 6.
CASC9 sponges miR-758-3p to promote EMT in BC by upregulating TGF-β2. (A) After knocking down the expression of CASC9, the expression changes of EMT-related genes were detected by the Human EMT RT2 Profiler PCR Array. (B) Target genes of CASC9 predicted by miRDB, TargetScan and LncBase. (C) TGF-β2 is a direct target of miR-758-3p. Upper panel, schematic diagrams of the mutual interactions between miR-758-3p and TGF-β2 3′UTR. Lower panel, luciferase reporter assay was performed to examine the effect of CASC9 on antagonizing miR-758-3p-mediated suppression of TGF-β2 expression. (D) CASC9 was transfected into BC cells (J82 and 5637), and TGF-β2 mRNA levels were evaluated by RT-qPCR. (E) Morphological changes of 5637 and J82 after knocking down CASC9 levels. (F and G) After transfection with CASC9, the protein levels of TGF-β2 and EMT markers were evaluated by western blotting. Unpaired Student's t-test was used and data are presented as the mean ± SEM. *P<0.05. CASC9, cancer susceptibility candidate 9; miR-758-3p, microRNA-758-3p; EMT, epithelial-mesenchymal transition; BC, bladder cancer; TGF, transforming growth factor; RT-qPCR, reverse transcription-quantitative PCR; SEM, standard error of mean; si, small interfering; NC, negative control.