Figure 8.

SMAD4 has a crucial role in miR-224–induced phenotypes. (a) Cell proliferation assay for SMAD4 and miR-224 overexpression in U2OS and MG-63 OS cells. The cell growth rates were measured by the MTT assay (n=4, mean±S.D.). (b) SMAD4 reduced the migration ability induced by miR-224 overexpression. Cell migration assay of miR-224-overexpressing cells transfected with SMAD4 or empty vector (n=4, mean±S.D., *P<0.01). (c) pre-miR-224 and MG-63 NC cells were treated with or without TGF-β for 8 h. Nuclear proteins were extracted and subjected to western blot analysis. (d and e) pre-miR-224 and MG-63 NC cells were treated with or without TGF-β for 8 h. The growth inhibitory effect and migration ability were measured by (d) cell proliferation assay and (e) migration assay. The values represent the means±S.D., *P<0.01. (f) Cell proliferation assay for SMAD4 and miR-224 knockdown in U2OS and MG-63 OS cells. The cell growth rates were measured by the MTT assay (n=4, mean±S.D.). (g) MiR-224 knockdown had no effect on the migration inhibition ability induced by SMAD4 knockdown. Cell migration assay of SMAD4 knockdown cells transfected with miR-224 sponge or control vector (n=4, mean±S.D., *P<0.01). (h) Nude mice were injected with 1 × 10⁷ MG-63 stable cells. Tumors were dissected and photographed after 5 weeks