Novel application of the published kinase inhibitor set to identify therapeutic targets and pathways in triple negative breast cancer subtypes
Fig 5
The pyrazolopyridazine GSK198271A promotes MET.
(A) Western blot analyses of MDA-MB-231 cells after treatment with GSK198271 confirms CDH1 protein expression in increased. Full length western blots are found in S3 Fig. Cells were pre-treated for 72 hours at 1μM; data was normalized to Rho. (B) Treatment of PDX tumors ex vivo with GSK198271 resulted in increased the epithelial marker CDH1 mRNA expression and reduced expression of CDH2 and MMP2, mesenchymal markers. Tumor pieces were treated for 72 hours at 1μM. (C) GSK198271A and not GSK350559A suppresses migration in another TNBC cell line, BT549. (D) Neither GSK198271A nor GSK350559A significantly increase CDH1 expression in BT549 cells. (E) GSK198271A and GSK350559A affected cellular proliferation at drug concentrations greater than 1μM. MDA-MB-231 cells were treated for 72 hours, then crystal violet stained and imaged. Absorbance was measured at 570 nm to obtain dose-response curves. Data was normalized to DMSO and performed in triplicate. (F,G) Representative images are enlarged for 1μM concentrations. Images were taken at 200x magnification using the Nikon color microscope. For qRT-PCR experiments, N = 3, error bars represent SEM and significantly different * p < 0.05, *** p < 0.001.