Figure 2.
TGF-β1 promotes cellular proliferation of meningioma cells in a dose-dependent manner. (a) IOMM-Lee and CH157-MN cells were grown to 90% confluence and then stimulated with TGF-β1 at various doses for 24 h. Cells were trypsinized and 0.4 × 105 cells were allowed to invade Matrigel-coated transwell inserts for 24 h at 37 °C. Then, the cells were fixed and stained. Invaded cells were photographed under a light microscope and the number of invaded cells was plotted against the dose of TGF-β1 (Columns: mean, bar: ±S.D.; n=3). (b) Cell lysates from IOMM-Lee and CH157-MN cells that were treated with the indicated doses of TGF-β1 for 48 h were subjected to western blot analysis for the detection of uPA, MMP-2, and MMP-9 using specific antibodies. GAPDH served as a loading control. Each blot is indicative of three independent experiments. (c) The band intensities from each blot were quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA) and relative expression was plotted. Columns represent mean±S.D. of three independent experiments. *P<0.05, significant difference from control without treatment. (d) Cell proliferation of meningioma cells that were treated with the indicated doses of TGF-β1for 72 h was measured using BrdU (colorimetric) kits. Data shown are mean±S.D. from three different experiments. *P<0.05, significant difference from control without treatment. (e) Cell lysates from IOMM-Lee and CH157-MN cells that were treated with specified doses of TGF-β1 for 48 h were subjected to western blotting for the detection of Bcl-XL, Cdc42, Cyclin D, P53 and P21 using specific antibodies. GAPDH served as a loading control. Each blot is indicative of three independent experiments. (f) The band intensities from each blot were quantified using ImageJ software, and the relative expression was plotted. Columns represent mean±S.D. of three independent experiments. *P<0.05, significant difference from control without treatment