Fig. 7

Radioresistance, proliferation, and EMT regulated by miR-151a-3p. Twenty-four hours after miR-151a-3p was transfected into HepG2, expression of p53 and p21, Puma, and Bcl-2 was confirmed by western blotting (A). Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. miR-NC by Student’s t-test. Cell cycle arrest assay using FACS was performed with PI staining to confirm that cell cycle arrest induced by radiation exposure was modulated by miR-151a-3p. Cell cycle was analyzed by dividing it into G1, S, and G2/M phases (B). Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. Non-radiated by Student’s t-test. To confirm that cell apoptosis induced by RT is modulated by miR-151a-3p, apoptosis was analyzed by Annexin V (FITC) and PI staining using FACS (C). Data are shown as mean ± SEM. *P < 0.05 vs. Non-radiated by Student’s t-test. Regulation of the representative epithelial-mesenchymal transition (EMT) markers E-cadherin, Twist1, Vimentin, and N-cadherin by miR-151a-3p was confirmed by western blot (D). Data are shown as mean ± SEM. *P < 0.05, **P < 0.01 vs. miR-NC by Student’s t-test. The cell proliferation according to the concentration of the miR-151a-3p was analyzed using CCK-8 assay (E). Data are shown as mean ± SEM. *P < 0.05 vs. Control by Student’s t-test. Cell migration and invasion ability according to the concentration of miR-151a-3p were analyzed using transwell assay. Five locations of each well were randomly photographed, and the migration and invasion were measured and compared with that of the control (F). Cell culture dishes were performed in at least three separate experiments. Scale bar, 100 µm. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01 vs. control by Student’s t-test