Circulating MiR-125b as a Marker Predicting Chemoresistance in Breast Cancer
Figure 6
E2F3 was a direct target of miR-125b in breast cancer cells.
(A) MCF-7 cells were transfected with 100 nM pre-miR-125b or pre-scramble control (Ctrl). (B) MDA-MB-231 cells were transfected with 100 nM anti-miR-125b or anti-scramble control (Ctrl). Twenty-four hours after transfection, nuclear lysates were prepared for Western blotting with an antibody against E2F3, and TBP was used as a loading control. (C) MCF-7 cells were co-transfected with luciferase reporter plasmids with intact or mutant 3′UTR of E2F3, pre-miR-125 or pre-scramble control (Ctrl). Luciferase activity was measured at 48 h post-transfection using a dual luciferase reporter assay. The results were expressed as relative luciferase activity (firefly/renilla luciferase). Data represented mean ± SD of triplicate experiments. *p = 0.002.