Fig. 3

miR-193a-5p inhibits PC3 cell apoptosis by increasing HO-1 expression. a PC3 cells were transfected with the indicated RNA constructs and then treated with H₂O₂, HO-1, p47^phox and p22^phox expression was determined by Western blotting. b Immunohistochemical staining of HO-1 at different stages of PC progression. 1, Benign prostatic hyperplasia (BPH); 2, Gleason grade 2; 3, Gleason grade 4; 4, Gleason grade 5 PC. HO-1 expression increases with high Gleason score. Bars = 40 μm. c Fluorescence in situ hybridization (FISH) staining was performed on sections from BPH and PC tissues. Green, red and blue staining indicates miR-193a-5p, HO-1 and DAPI, respectively. Bar = 64 μm. d HO-1 mRNA and miR-193a-5p were measured by RT-qPCR, and Pearson correlation analysis shows a positive correlation between miR-193a-5p and HO-1. (P = 0.0157; R = 0.5325). e PC3 cells were treated with the different concentrations of Docetaxel (Doc), and the expression of miR-193a-5p was detected by qRT-PCR. *P < 0.05, **P < 0.01 vs. vehicle control. f Following transfection with the indicated RNA constructs, PC3 cells were treated with Doc (10 nM). HO-1 and cleaved caspase-3 were determined by Western blotting. g PC3 cells were treated with Doc (10 nM) alone or together with 20 μM Hemin (sigma, #51280) or Znpp (Protoporphyrin IX zinc, sigma, #282820) for 24 h, and cell apoptosis was assessed by flow cytometry. h PC3 cells were treated as in (G), TUNEL staining detected cell apoptosis. Right panel shows the number of TUNEL-positive cells of three independent experiments. Bar = 100 μm. *P < 0.05 vs. Doc alone; **P < 0.01 vs. vehicle control. i PC3 cells were treated as in (g), Western blotting detected HO-1 and cleaved caspase-3. Experiments were repeated three times