Study On The Mechanism Of Disulfiram Inhibits Epithelial-Mesenchymal Transition In NSCLC Cells

OBJECTIVE: Epithelial-mesenchymal transition is an important cause of invasion and metastasis and chemotherapy tolerance in non-small cell lung cancer.Inhibition of epithelial-mesenchymal transition has become an important strategy for the treatment of non-small cell lung cancer and drug resistance.Although there are many studies on epithelial-mesenchymal transition,there is still a lack of effective interventions.Recent studies have shown that ALDH1 is an important marker in the process of epithelial-mesenchymal transition,and its blocking drug disulfiram has also been shown to inhibit tumors.This study aimed to explore the inhibitory effect of disulfiram on TGF-β1-induced EMT in non-small cell lung cancer,in order to provide more strategies for clinical treatment of NSCLC.METHODS: 1.TGF-β induced A549 cells to construct EMT model.2.MTT assay to detect the effect of disulfiram on TGF-β1-induced NSCLC proliferation in epithelialmesenchymal transition.3.Inhibition of migration,adhesion and invasion of A549cells by EMT was determined by cell scratch,matrix adhesion,endothelial cell adhesion,and transwell chamber assay.The effect of DSF on the expression of Ecadherin and Vementin in EMT of A549 cells was analyzed by western blot,and the effect of DSF on vimentin and E-cadherin mRNA was detected by q PCR.5.The expression levels of mRNAs of EMT-related stem cell marker genes ALDH-1,SOX2 and OCT4 were detected by q PCR.RESULTS: 1.TGF-β1 can effectively induce EMT in A549 cells,which is transformed from epithelial to mesenchymal morphology,and DSF can reverse the morphological changes induced by TGF-β1.2.DSF inhibited the proliferation of A549 cells with EMT in a concentration-dependent manner.3.TGF-β1-induced A549 after EMT,its metastatic biological behavior was enhanced,scratch test,matrix adhesion,endothelial cell adhesion,transwell chamber experiments,the results showed that DSF induced TGF-β1-induced A549 after EMT The enhancement of migration,adhesion,and invasive ability has obvious inhibitory effects and is dose dependent.TGF-β1 induced the expression of the interstitial phenotype protein Vementin in A549 cells and decreased the expression of epithelial phenotype protein E-cadherin.DSF significantly decreased the expression of Vementin and promoted the expression of E-cadherin.5.DSF can significantly reduce the mRNA expression levels of stem cell marker genes ALDH-1,SOX2 and OCT4 induced by TGF-β1 in E549 cells.CONCLUSION: DSF can inhibit the proliferation of tumor cells after EMT,which can reverse the EMT process of A549 cells induced by TGF-β1,thereby inhibiting cell invasion and migration.Under the action of DSF,the related proteins in A549cells can change,and the tumor cell dry related genes ALDH-1,SOX2 and OCT4 can be down-regulated by DSF.