Fig. 4

CircBNC2 regulates ACSL6 expression by sponging in PCa cells. A Venn diagram showing 387 potential target genes which predicted by four databases (TargetScan, miRPathDB, miRWalk, and mirDIP). B Volcano plot showing the profile of differentially expressed genes based on TCGA of PCa. C Correlation analysis revealed negative correlation between the levels of ACSL6 and miR-4298 in 78 PCa patient samples. D The schematic illustration for construction of luciferase reporter plasmids, wild-type (WT) and mutant (Mut)-ACSL6, as indicated (left panel). The luciferase activity was measured in DU145 and PC3 cells which co-transfected with of the luciferase reporter plasmid WT/Mut-ACSL6 and miR-4298 mimic/NC mimic, respectively (right panel). E - H The expression level of ACSL6 was analyzed in DU145 and PC3 cells by RT-qPCR and western blotting after transfection with NC mimic/miR-4298 mimic (E and F) or NC inhibitor/miR-4298 inhibitor (G and H), respectively. I - J The CCK-8 assay (I) and transwell assay (J) showing that miR-4298 mimic or NC mimic affected the proliferative and invasive ability of OE-ACSL6 DU145 and PC3 cells. K and L The expression level of ACSL6 was validated by RT-qPCR (K) and western blotting (L) in OE-circBCN2 DU145 and PC3 cells. M In OE-circBNC2 PC3 and DU145 cells, western blotting analyzed the ACSL6 expression upon miR-4298 mimic or NC mimic treatment. N and O CCK-8 assay (N) and transwell assay (O) showing that sh-ACSL6 knockdown reverse the inhibition of proliferation and invasion in PC3 and DU145 cells caused by circBNC2 overexpression. P - R The representative images of subcutaneous xenograft tumors (n = 6/group) (P), tumor growth curves (Q), and tumor weight (R) at the endpoint from control (OE-Ctrl) and circBNC2 overexpression (OE-circBNC2) groups co-treated with knocking down of control (sh-Ctrl) or knocking down of ACSL6 (sh-ACSL6). S Correlation analysis revealed positive correlation between the levels of ACSL6 and circBNC2 in the 78 PCa patient samples. Scale bar = 50 μm. ns, no significance; *P < 0.05; **P < 0.01; ***P < 0.001