Figure 5.

miR-148a directly targets PTEN expression in osteosarcoma. (A) Sequence alignment of miR-148a and its predictive target sites in the 3′-UTR region of PTEN. (B) MG63 cells were co-transfected with PTEN 3′-UTR firefly luciferase reporter plasmid, or with a reporter construct whereby the miR-148a target sites had been deleted, together with Renilla luciferase plasmids, control RNA or miR-148a mimics, as indicated. Following 48 h, luciferase activity was measured and normalized. (C) MG63 and U2OS cells were transfected with miR-148a mimics or miR-148a inhibitor, as indicated. Protein expression of PTEN and β-actin (which served as an internal control) were detected by western blotting. Data are presented as the mean ± standard deviation (n=4) of one representative experiment. Similar results were obtained in three independent experiments. **P<0.01. miR, microRNA; Ctrl, control; UTR, untranslated region; PTEN, phosphatase and tensin homolog.