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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Bazedoxifene as a novel GP130 inhibitor for Colon Cancer therapy

Fig. 4

Bazedoxifene inhibits induction of STAT3 phosphorylation and cell proliferation by IL-11. a: DLD-1, HCT-116, and HCT-15 cells were starved in serum-free medium for 24 h and pre-treated with bazedoxifene (5~20 μM) for 2 h. Then, 50 ng/ml (DLD-1 cells) or 25 ng/ml IL-11 (HCT-116 and HCT-15 cells), 50 ng/ml OSM (DLD-1 cells) and 50 ng/ml IFN-γ (DLD-1 cells) were added for stimulation. The p-STAT3Y705, p-STAT1Y701, STAT3, STAT1 and GAPDH were assessed by western blot analysis. b: DLD-1 cells were starved in serum-free medium for 24 h and pre-treated with bazedoxifene (10 μM) for 2 h followed by IL-11 stimulation (50 ng/ml). STAT3 nuclear translocation was detected by immunofluorescence. c: 3000 cells per well of DLD-1, HCT-116, and HCT-15 were seeded in 96-well plates and starved in serum-free medium for 24 h. The next day, cells were pretreated with 5 μM bazedoxifene for 2 h alone or followed with 50 ng/ml IL-11 stimulation. After 24-h treatment, cell viability was determined by MTT assay. d: Induction of cell proliferation and p-STAT3 is inhibited by bazedoxifene in colon cancer cells. 8000 cells per well of DLD-1, HCT-116 and HCT-15 cells were seeded in 96-well plates and then starved in serum-free medium for 24 h. Pretreatment with 5~15 μM bazedoxifene for 4 h was followed by 10 ng/ml IL-11 stimulation for 24 h. BrdU assay was performed as described previously. e: IL-11Rα is knocked down in DLD-1, HCT-116, and HCT-15 cells. Cells were transfected with 10 nM of negative control siRNA or human IL-11Rα siRNA. After 48 h, cells were harvested and lysed for western blot or processed for MTT cell viability assay. Cells were then treated with bazedoxifene for another 72 h. IL-11R was assessed by western blot analysis with GAPDH as a control. Cell viability was determined by MTT assay. (*, p < 0.05; **, p < 0.01; ***, p < 0.001)

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