Figure 6
Inhibition of miR-301a reduces tumor growth in mouse xenografts through SMAD4. (A) 2 × 105 transformed BEAS-2B (BEAS-2B-As) with Anti-miR-control or Anti-miR-301a were injected subcutaneously into nude mice. Representative images of tumor from nude mice are shown. (B) Tumor volume of each group was measured respectively. (C) Detecting the expression of SMAD4 in tumors of each group. Scale bars = 200 μm. (D) 2 × 105 transformed BEAS-2B (BEAS-2B-As) with Anti-miR-301a plus either shRNA control or shRNA-Smad4 were injected subcutaneously into nude mice. Representative images of tumor from nude mice are shown. (E) Tumor volume of each group from (D) was measured respectively. (F) Western blot analyses of SMAD4, phospho-Stat3 (pStat3) and total Stat3 of tumors from the indicated groups. Gel pictures were acquired by using Tanon5200 and the exposure time was 30 sec for SMAD4, 60 sec for pStat3, 30 sec for Stat3 and 10 sec for β-actin. (G) BEAS-2B cells were transfect with the indicated shRNA control or shRNA-Smad4 in the presence with LNA-anti-miR-301a(Anti-miR-301a). After 48 h post transfection, cells were treated with arsenic (5 and 10 μM) for 24 h and western blot analyses of phospho-Stat3 (pStat3) and Stat3 expression. Gel pictures were acquired by using Tanon5200 and the exposure time was 60 sec for p-Stat3, 30 sec for Stat3 and 10 sec for β-actin. Values represented the mean ± SD (n = 5 mice per group). *P < 0.05 and **P < 0.01 indicate a significant difference between the indicated groups.
