Fig. 4

CDK6 is the direct target and down regulated by miR-500a-3p. A Potential binding sites predicted by Targetscan and miRDB between miR-500a-3p and CDK6 mRNA. B CDK6 mRNA levels after transfected with miR-500a-3p in HCT116 and SW480 cells by qPCR. The data were presented as the means ± SD (n = 3). C CDK6 protein levels after transfected with miR-500a-3p in HCT-116, SW480 and HT29 cells by western blot. D Schematic representation of luciferase reporter construct containing CDK6 3′ UTR with either wild type (WT) or mutant (MT) miR-500a-3p target site. Binding sites were marked in red. E, F The luciferase activity was detected in SW480 cells co-transfected with CDK6-WT or CDK6-MT and miR-NC or miR-500a-3p. The data were presented as the means ± SD (n = 3). G The specimens in the cohort which contained 134 continuous CRC patients were stained with CDK6. Scale bar, 100 μm. H Association between miR-500a-3p mRNA level and CDK6 IHC score were conducted in the cohort (n = 134) (Spearman’s correlation analysis). **P < 0.01, ****P < 0.0001