Fig. 6.

MiR-193a directly targets HOTAIR and negatively modulates its expression in PCa. a-b HOTAIR expression was measured by qRT-PCR in PC3 and DU145 cells with overexpression of miR-193a by transfection of miR-193a mimics or infection with LV-miR-193a. GAPDH were used as internal controls. c-d Expression of miR-193a and HOTAIR were measured by qRT-PCR in miR-193a-overexpressing PC3 xenograft tumor tissues. U6 snRNA and GAPDH were used as internal controls for miR-193a and HOTAIR, respectively. e Schematic showing HOTAIR contained a complementary site to the seed region of miR-193a (551-577 bp) predicted by miRcode and DIANA Tools. f The luciferase activity of modified psiCHECK-2 luciferase reporter vector containing wild type or mutations of the binding sites of HOTAIR cDNA. MiR-193a mimics or miR-NC was co-transfected into PC3 and DU145 cells for 48 h. HOTAIR mut as negative control. Rluc activity in the cells was measured and normalized to Fluc activity. g-i ISH and IHC staining on 31 PCa clinical tissues with miR-193a/HOTAIR probes and an anti-EZH2 antibody. Spearman correlation analysis showing inverse correlation between HOTAIR and miR-193a as well as between EZH2 and miR-193a. The original magnification: ×200. Each bar represents the mean ± SD of three independent experiments. *P < 0.05