Figure 4

miR-494-3p represses PTEN expression and activates PI3K in human hepatocellular carcinoma cells. (A) Sequences showing the putative miR-494-3p binding sites on PTEN 3′UTR and the mutated miR-494-3p binding sites generated by site-directed mutagenesis. (B) In SMMC7721 and (C) HCCLM3, dual luciferase reporter assay showing significant suppression of luciferase activity only when miR-494-3p was partnered with wild-type PTEN 3′UTR. (D) Western blot analysis of PTEN protein expression after transfection in SMMC7721 and HCCLM3 cells. (E,F) The migratory properties of the cells were analyzed using the Transwell migration assay with Transwell filter chambers. Results are plotted as the average number of migrated cells from 6 random microscopic fields and the invasive properties of the cells were analyzed with the invasion assay using BioCoat Matrigel invasion chambers. Results are plotted as the average number of invasive cells from 6 random microscopic fields. (*p < 0.05, **p < 0.01, and ***p < 0.001).