Effect Of Tetrandrine On Proliferation,Migration,Invasion And EMT Of Human Pancreatic Cancer Cells Via Smad2/3/MAPK Pathway

Objective: This study was conducted to investigate the effect of tetrandrine on proliferation,migration and invasion of human pancreatic cancer cells CFPAC-1 and PANC-1 by in vitro experiments,and to explore the molecular mechanism of its action.Methods: Firstly,human pancreatic cancer cells CFPAC-1 and PANC-1 were cultured in vitro.CCK-8 assay and cell clone formation assay were used to detect the effects of different concentrations of tetrandrine on cell viability and proliferation of human pancreatic cancer cells.Wound healing assay and Transwell migration and invasion assays were used to detect the effects of different concentrations of tetrandrine on the migration and invasion of cells.Western blot assay was used to detect the effect of tetrandrine on the expression levels of EMT(epithelial mesenchymal transition)related proteins E-cadherin,Ncadherin and Vimentin and their upstream transcription factors Twist and Snail1 in CFPAC-1 and PANC-1 cells.The expression levels of key proteins Smad2,pSmad2,Smad3 and p-Smad3 in TGF-?/Smad pathway and the expressions of ERK,p-ERK,P38 and p-P38 proteins in MAPK pathway were all detected by western blot assay.Results: 1.Different concentrations of tetrandrine(0?M,5.0?M,10.0?M,20.0?M,40.0?M,60.0?M)inhibited the proliferation of human pancreatic cancer cells,and this effect was time-dependent and dose-dependent(p < 0.05).2.Through cell scratch test and Transwell experiment,it was found that tetrandrine can inhibit the migration and invasion of human pancreatic cancer cells.After interfering with CFPAC-1 cells and PANC-1 cells with tetrandrine(0?M,5?M,20?M)for 24 h,cells were used.The migration distance,and the number of cells passing through were significantly less than the control group,and were concentration-dependent(p < 0.05).3.Western blot results showed that human pancreatic cancer cells CFPAC-1 and PANC-1 were treated with tetrandrine(0?M,5?M,20?M)for 24 h,and the EMT-related proteins and upstream key transcription factors of the cells changed.The expression level of E-cadherin was increased in cells,and the expression levels of N-cadherin,Vimentin,Snail1 and Twist were decreased.At the same time,the key proteins in the TGF-?/Smad and MAPK signaling pathways of EMT regulation also changed.The expression levels of total Smad2,Smad3,ERK and P38 in the cells did not change significantly,but their expression levels of p-Smad2,p-Smad3,p-ERK,and p-P38 were significantly decreased(p <0.05).Conclusion: Tetrandrine can inhibit the proliferation,migration,invasion and EMT of human pancreatic cancer cells CFPAC-1 and PANC-1.And,tetrandrine inhibits EMT of pancreatic cancer cells by affecting TGF-?/Smad and MAPK signaling pathways.Therefore,the inhibitory effect of tetrandrine on human pancreatic cancer cells can provide new ideas for the treatment of pancreatic cancer.