Fig. 2
From: Metformin reverses mesenchymal phenotype of primary breast cancer cells through STAT3/NF-κB pathways
Metformin reduces the expression of Vimentin and SNAIL, decreases cell proliferation but not migration in mesenchymal breast cancer cells. a Primary breast cancer cell with mesenchymal phenotype (MBCDF-D5, MBCD3, MBCDF-B3 and MBCD23) were treated with 10 mM MTF for 0, 1 and 2 days and the effect of MTF on the mesenchymal markers Vimentin and SNAIL was analyzed by immunoblotting. Actin was used as loading control. b For cell proliferation, primary breast cancer cells with mesenchymal phenotype (MBCDF-D5, MBCD3, MBCDF-B3 and MBCD23) were seeded at 2500 cell/cm2 (5000 cells/well) in a 24 well-plate and incubated in the absence (control) or presence of 0, 5,10, and 25 mM MTF. Cell proliferation was evaluated by MTT at days 0, and 6. Data represents the mean ± SEM of three independent experiments performed in triplicate incubations. *P < 0.05. c Migration assays were performed using Boyden chambers. Thirty thousand cells with mesenchymal phenotype (MBCDF-D5, MBCD3, MBCDF-B3 and MBCD23) were seeded in the upper chamber in presence of MTF 0, 10 and 25 mM, the same concentrations of MTF were added in the in-bottom chamber, and then incubated for 6 h at 37 °C. After this time the cells that did not migrate were removed from the upper chamber. Cells that migrated were fixed and stained with Cristal Violet. Five fields were counted under the microscope at 20X. Migration assays were performed three independent times in triplicate