Fig. 4

MiR-142-3p suppressed the cell proliferation and promoted cell apoptosis by targeting SMAD5 in hepatocellular carcinoma. a Schematic diagrams of the putative interactions between miR-142-3p and SMAD5. b The expression of SMAD5 in hepatocellular carcinoma tissues and adjacent non-tumor tissues was assessed by Q-PCR. c Q-PCR was used to analyze the mRNA level of SMAD5 in 5 hepatocellular carcinoma cell lines. d HepG2 cells were transfected by miR-142-3p mimic control mimic (NC), and the level of miR-142-3p was analyzed by Q-PCR. e The HepG2 and SMMC-7721 cells were transfected by miR-142-3p mimic control mimic (NC); the mRNA was analyzed by Q-PCR. f HepG2 and SMMC-7721 cells were transfected by miR-142-3p mimic control mimic (NC). The protein level of SMAD5 was analyzed by Western blot. g The luciferase reporter plasmid containing wild-type or mutant SMAD5 was co-transfected with miR-142-3p mimics or control mimic into HEK-293 T cells. Luciferase activity was analyzed and normalized to Renilla activity. h SMAD5 in HepG2 and SMMC-7721 cells was ectopically expressed using a plasmid containing SMAD5. The plasmid vector pcDNA3.0 was used as the control. The miR-142-3p mimic control mimic (NC) was also used as indicated in the figure. Cell viability was assessed by CCK-8 assay after the indicated transfection in HepG2 and SMMC-7721 cells. i The cell proliferation of HepG2 and SMMC-7721 cells after the indicated transfection was assessed by EdU assay. Representative images of the cells are shown. Scale bar, 100 μm. j Cell apoptosis of HepG2 and SMMC-7721 cells after the indicated transfection was assessed by flow cytometry. Data represent three independent experiments (mean and SEM of triplicate samples). *P < 0.05, **P < 0.01. ***P < 0.001. SEM, standard error of the mean