Figure 3
E 2 -responsive miRNAs downregulated endogenous bcl-2, cyclin D1 and survivin expression. (A) MCF-7 cells were transfected with 40 nM miR-16, miR-143 and miR-203 mimics for 48 h, Western blot was performed to detect endogenous bcl-2, cyclin D1 and survivin expression. (B) The densitometry of each gene vs. β-actin was indicated and statistically analyzed. * denotes P < 0.05 compared with negative control miRNA. (C) RNA was extracted from MCF-7 transfected with miRNA mimics and RT-QPCR was performed to confirm the overexpression of these miRNAs. (D, E) MCF-7 cells were transfected with 40 nM miR-16, miR-143 and miR-203 inhibitors. After 48 h, Western blot was performed to examine endogenous survivin expression. Meanwhile, RT-QPCR was performed to examine the miRNAs expression. (F) MCF-7 cells were co-transfected with 40 nM miR-16, miR-143 and miR-203 mimics together with 100 ng bcl-2, cyclin D1 and survivin 3'UTR luciferase constructs and 10 ng pRL-SV40. After 24 h, luciferase activity assay was determined. * denotes P < 0.05 compared with negative control.