Figure 2.

MiR-326 was targeted by circHIPK3 in MCF7 cells. (a) Cytoplasmic and nuclear RNA fractions were isolated from MCF7 cells. Relative expressions of circHIPK3 in the cell cytoplasm or nuclei were examined by RT-qPCR. GAPDH was used as the cytoplasmic control, and U6 was used as a nuclear control. (b) The binding sites for miR-326 in circHIPK3 were predicted by circInteractome. (c) Luciferase reporter assay showed that miR-326 inhibitor promoted the luciferase activity of circHIPK3 in MCF7 cells. (d) Relative expression of miR-326 was detected after transfection with sicircHIPK3 in MCF7 cells by RT-qPCR. U6 was used as an internal control. (e) Relative miR-326 expression was detected in the BCa tissues and adjacent non-cancerous tissues (ANT) was determined by RT-qPCR. (f and g) The correlation between circHIPK3 and miR-326 in BCa tissues and ANT was determined by Pearson’s correlation analysis. ***P < 0.001 vs. Cytoplasm; ^#P < 0.05 vs. IC; ^{^^}P < 0.01 vs. siNC; ^&&& P < 0.001 vs. ANT. Data are shown as mean±SD, n = 3. Mut, mutant; WT, wild-type; siCircHIPK3, small interfering circular RNA HIPK3; siNC, small interfering negative control; IC, inhibitor control. RT-qPCR, reverse transcription-quantitative polymerase chain reaction