Figure 4: miR-760 directly targets IL6 by binding to its 3’UTR. (a) The putative binding site of miR-760 in the 3’UTR of IL6. The mutated site is also shown in the diagram. (b) Control or miR-760 mimics were cotransfected with psiCHECK reporters containing the wild-type or mutated 3’UTR of IL6. Luciferase activity was measured as indicated. (c) qPCR analysis of IL6 in LNCaP cells that were transfected with control inhibitor or miR-760 inhibitor or transfected with conrol mimics or miR-760 mimics. siNC or siIL6 LNCaP and 22rv1 cells were subjected to (d) western blot analysis of IL6 and (e) CCK analysis of cell proliferation. (f) qPCR analysis of IL6 levels in prostate cancer and adjacent normal tissues (n = 10). (g) qPCR analysis of IL6 levels in prostate cancer(n = 18) and nonpaired normal (n = 10) tissues. *P < 0.05, **P < 0.01, ***P < 0.001. NS: no significance. IL6: interleukin 6; CCK: Cell Counting Kit; NC:negative control; qPCR: quantitative polymerase chain reaction; WT: wide type; MU: mutant.