Disulfiram Inhibits TGF-?-induced Epithelial-mesenchymal Transition In Breast Cancer

?Purpose? To investigate the effect of DSF on TGF-?-induced EMT in breast cancer cell lines MCF-7 and MDA-MB-231,and explore the possible mechanism of the phenomenon.?Method? 1.MTT assay was performed to measure the effect of disulfiram on cytotoxicity of MCF-7 and MDA-MB-231 cell lines,and the DSF working concentration was selected according to the MTT result.2.The experimental groups were divided into control group,TGF-? group,and different concentrations of DSF combined with TGF-? group.3.To observe the inhibitory effect of DSF on TGF-?-induced EMT: The morphology of the cells was observed by light microscopy,and the expression of EMT related markers(i.e.E-cadherin,N-cadherin,Vimentin)were detected by immunofluorescence and Western blot.4.The effect of DSF on the migration and invasion ability of breast cancer cells were detected by cell scratch assay,transwell chamber migration assay and Matrigel invasion assay.5.The expression of NF-?B,ERK,p-ERK and Snail were detected by western blot to demonstrate the inhibitory effect of DSF on ERK / NF-?B / Snail pathway.U0126,the inhibtior of ERK,was used on breast cancer cells to demonstrate whether DSF could inhibite EMT by inhibiting the ERK / NF-?B / Snail pathway.?Result? 1.DSF inhibited the proliferation of MCF-7 and MDA-MB-231 cells,and the inhibitory rate increased with the increase of concentration of DSF.Moreover,the inhibition rate was below 25% in the experimental concentration,indicating that the effect of DSF on the following function was not caused by cytotoxicity.2.TGF-? induced EMT in breast cancer cells,and the cell morphology became from the typical polarity,cobble-shaped epithelial cells to non-polar,spindle-like stromal cells.However,with the increase of the concentration of DSF,the cell morphologgy gradually changed to epithelial morphological type in the DSF combination with TGF-? group.TGF-? increased the expression of Vimentin /N-cadherin,but inhibited the expression of E-cadherin,and this effect could be suppressed by DSF in a dose-dependent manner.3.In the invasion and migration experiment,TGF-? promoted cell invasion and migration ability,but this effect was inhibited by DSF.4.The expression of NF-?B / P65 in TGF-? group was up-regulated and the expression of ERK and Snail was enhanced,while DSF significantly inhibited NF-?B activity and down-regulated ERK / NF-?B / Snail pathway.5.U0126,the inhibtior of MEK1/2,inhibited ERK / NF-?B / Snail pathway,then it inhibited EMT in breast cancer cells.?Conclusion? 1.DSF inhibited TGF-?-induced EMT in breast cancer cells.2.DSF inhibited the invasion and migration abillity which promoted by TGF-? in breast cancer cells.3.DSF inhibited NF-?B activity and down-regulated ERK / NF-?B / Snail pathway.4.Blocking ERK / NF-?B / Snail pathway could inhibit TGF-?-induced EMT breast cancer cells.5.In breast cancer cells,DSF inhibited TGF-?-induced EMT by inhibiting ERK / NF-?B / Snail pathway.