Table 1.
Sequences of miR-182 mimics and inhibitors.
Figure 1.
Significantly different expressed miRNAs between PCa and BPH by miRNA microarray analysis.
In PCa tissues, five miRNAs were down-regulated and twenty-two miRNAs were up-regulated. miRNA-182 was up-regulated with fold change of 3.07 and further confirmed by real-time RT-PCR with fold change of 2.85.
Figure 2.
miR-182 expression in different prostate cells measured by real-time RT-PCR.
Real-time RT-PCR show that the relative expression level of PC-3 cells is the highest in four prostate cancer cell lines and RWPE-1 has the lowest expression level. Values represent means from three separate experiments and error bars represent the SD.
Figure 3.
PC-3 cells were observed using fluorescence microscope after transfection with miR-182 mimics or inhibitors.
Results show that the transfection rate is more than 80%. Representative images and randomly selected fields are shown.
Figure 4.
MiR-182 expression in PC-3 cells measured by real-time RT-PCR after transfection with miR-182 mimics or inhibitors.
Blank presents blank control. The results show that the expression level is highest in miR-182 mimics group and lowest in miR-182 inhibitor group. Values represent means from three separate experiments and error bars represent the SD.
Figure 5.
Viability of PC-3 cells transfected with miR-182 mimics or inhibitors was measured by MTT assays.
Blank presents blank control. UV-visible absorbance was measured at 490 nm. The relative absorbance was significantly different 3 days after transfection (P<0.05). Values represent means from three separate experiments.
Figure 6.
Analysis of cell cycle in PC3 cells after transfected with miR-182 mimics or inhibitors.
Blank presents blank control. Values represent means from three separate experiments and error bars represent the SD. (*P<0.05).
Figure 7.
Early apoptosis detection of PC-3 cells transfected with miR-182 mimics or inhibitors.
Blank presents blank control. The results show that the early apoptosis rate is lowest in miR-182 mimics group and highest in miR-182 inhibitor group (*P<0.05, **P<0.01). Representative images were shown and early apoptosis cells were indicated at Q4 (LR) area.
Figure 8.
Invasion assays were performed with PC-3 cells transfected with miR-182 mimics or inhibitors. Blank presents blank control. Representative images and randomly selected fields are shown (*P<0.05, **P<0.01).
Figure 9.
Schematic digram of NDRG1 3′-UTR region targeted by miR-182.
NDRG1 3′-UTR region has only one highly conserved miR-182 binding sites after bioinformatic analysis.
Figure 10.
Western Blotting results of NDRG1 protein expression in PC-3 cells after transfection with miR-182 mimics or inhibitors.
Blank presents blank control. Overexpression of miR-182 significantly decreased the protein expression levels of NDRG1 and downregulation of miR-182 dramatically increased the protein expression levels of NDRG1 (*P<0.05, **P<0.01). Values represent means from three separate experiments and error bars represent the SD.
Figure 11.
Relative luciferase activity of the pmirGLO/NDRG1-UTR in PC-3 cells.
Luciferase activity of the pmirGLO/NDRG1-UTR was dramatically inhibited in miR-182 mimics group and significantly increased in miR-182 inhibitors group, compared to the control groups (*P<0.05, **P<0.01).