Figure 4

PTEN is a functional target of miR-492 in HCCs . (A) HCC tissues and the adjacent non-cancerous tissues were subject to Western blot and real-time PCR analyses to assess PTEN mRNA and protein levels. Data are the mean ± SD (n = 4) of one representative experiment. Similar results were obtained in at least three independent experiments. (B) HEK-293 cells were transfected with either miR-492 mimics (20 nM) or the mutant 3′-UTR of miR-492 (20 nM) along with the matched control using 80 ng of indicated vectors and 40 ng of pRL-TK. After 24 h, firefly luciferase activity was measured and normalized by Renilla luciferase activity. Data are the mean ± SD (n = 4) of one representative experiment. Similar results were obtained in at least three independent experiments. The proliferation of HepG2 cells transfected with miR-492 plasmid (0.5 nM) and different concentrations of PTEN plasmid (5 nM, 10 nM, 20 nM) was determined by MTT. *indicates the significant difference between the control and PTEN transfected cells. (C) HCT116 or HepG2 cells were infected with the indicated lentivirus for 24 h, and p-AKT, total AKT and PTEN levels were detected by immunoblot analysis. Data shown is one representative experiment out of three. (D) miR-492 transfection enhanced the proliferation of both HCT116 and HepG2 which was inhibited by the PI3K and ATK inhibitors.