MicroRNA-195 Inhibits the Proliferation of Human Glioma Cells by Directly Targeting Cyclin D1 and Cyclin E1
Figure 4
miR-195 downregulates cyclin D1 and cyclin E1 by directly targeting their 3
′-UTRs. A, Predicted miR-195 target sequence in the 3′-UTR of cyclin D1 and cyclin E1 (cyclin D1 3′-UTR and cyclin E1 3′-UTR) and illustration of the three altered nucleotides in miR-195-mut. B, Western blotting analysis of expression of phosphorylated Rb (p-Rb), total Rb, PCNA, cyclin D1, cyclin E1, CDK2 and p21 in indicated cells. α-Tubulin served as the loading control. C, Western blotting analysis of GFP expression in indicated cells. D, Luciferase assay of indicated cells transfected with the pGL3-cyclin D1-3′UTR reporter (left) or the pGL3-cyclin E1-3′UTR reporter (right) with increasing amounts (10, 50 nM) of miR-195 mimic, or miR-195 inhibitor, or miR-195 mutant. Each bar represents the mean ± SD of three independent experiments. * P<0.05.