Eribulin rapidly inhibits TGF-β-induced Snail expression and can induce Slug expression in a Smad4-dependent manner

摘要

TGF-β causes rapid induction of Snail, which is differentially inhibited by microtubule targeting agents. Western blot analysis of lysates from BT-549 and HCC1937 cells that were serum-starved for 18 h then stimulated with TGF-β at the indicated times and concentrations. Immunoblots were probed for Snail and GAPDH. mRNA expression analysis of in BT-549 and HCC1937 cells. Cells were serum-starved for 18 h then stimulated with 2 ng/mL of TGF-β for indicated times. Data are an average of two independent experiments (±SEM). Western blot analysis of Snail and GAPH from whole-cell lysates from BT-549 and HCC1937 cells that were serum-starved for 18 h, then pre-treated with microtubule targeting agents for 2 h followed by a 3 h stimulation with TGF-β. mRNA analysis of transcript in cells that were serum-starved for 18 h, pre-treated with microtubule targeting agents for 2 h, then stimulated with TGF-β for either 1 h (BT-549 cells,  = 3) or 2 h (HCC1937 cells,  = 4). Data are the average of independent experiments (± SEM). A one-way ANOVA with Dunnett’s post-hoc test was used to determine statistical significance as compared to TGF-β-stimulated vehicle control cells (****  p p p e, . Smad2/3 proteins were transiently knocked-down by siRNA in BT-549 and HCC1937 cells after which cells were serum-starved for 18 h, pre-treated with microtubule targeting agents for 2 h, and then stimulated with TGF-β for 3 h. Whole-cell lysates were subject to immunoblotting and probed for Smad2/3, Snail, and GAPDH. , Western blot analysis of whole-cell lysates from cells serum-starved for 18 h, pre-treated with microtubule targeting agents for 2 h, then stimulated with TGF-β for either 45 min (BT-549) or 1 h (HCC1937). Immunoblots were probed for total Smad2/3, their phosphorylated forms, and GAPDH