Research Paper Volume 12, Issue 6 pp 5399—5410

The therapeutic value of the SphK1-targeting microRNA-3677 in human osteosarcoma cells

Figure 2. Ectopic overexpression inhibits OS cell progression in vitro. Sable OS-1 cells with the pre-miR-3677-expressing lentivirus (“lv-pre-miR-3677”, s-L1/s-L2, two lines) or with non-sense control miRNA (“lvmiC”), as well as the parental control OS-1 cells (“Ctr”), were cultured, with cell growth curve shown in (A); Cell colony formation (B), proliferation (EdU incorporation, C) and migration (“Transwell” assay, D) were tested by mentioned assays, with cell apoptosis examined by TUNEL staining (E) and Annexin V FACS (F) assays. U2OS cells and MG63 cells as well as primary human OS cells (OS-2 and OS-3) were infected with lv-pre-miR-3677 or lvmiC for indicated time periods, cell proliferation and apoptosis were tested by EdU incorporation (G) and TUNEL staining (H), respectively. For in vitro cell functional assays, the exact same number of viable cells with different genetic modifications were initially plated into each well/dish (at 0h/Day-0, same for all figures). Data were presented as mean ± SD (n=5), and results were normalized. ***P< 0.001 vs. “lvmiC”/“miC” cells. Experiments in this figure were repeated five times with similar results obtained. Bar=100 μm (CE).