Figure 4

Cyclin D1 and CDK6 are miR-211 targets. A. Schematic representation showing the five predicted miR-211 targets (TCF4, Cyclin D1, Cyclin D2, Cyclin D3 and CDK6) screened using three algorithms (miRNA.org, miRWalk and Targetscan). B. OVCAR3 cells were co-transfected with TCF4, CDK6, Cyclin D1, Cyclin D2 or Cyclin D3 3′UTR pGL plasmid with miR-211. Forty-eight hours later, luciferase assay was performed. *p < 0.05 compared to miR-Ctrl transfected cells. Data are presented as mean ± SEM of three independent experiments. C-D. Sequence of miR-211 with the putative binding sites of Cyclin D1 (C) and CDK6 (D) 3′ UTR. E. Western blot of Cyclin D1 and CDK6 levels in OVCAR3 and SKOV3 cells transfected with miR-211 or miR-Ctrl. F. Western blot of Cyclin D1 and CDK6 levels in OVCAR3 and SKOV3 cells transfected with LV-miR-211 or LV-miR-Ctrl. G-H. The mRNA expression levels of Cyclin D1 (G) or CDK6 (H) correlated inversely with miR-211. I-J. Cyclin D1 (I) and CDK6 (J) expression in ovarian tissues (GDS3592/Cyclin D1, n = 24, p = 0.017; GDS3592/CDK6, n = 24, p = 0.016).