Effect Of Plumbagin On Epithelial-mesenchymal Transition In Hepatocellular Carcinoma Cells And Its Mechanism

Objective: The object of this study was SMMC-7721 hepatocellular carcinoma(HCC)cell lines,To investigate the effect of plumbagin on transforming growth factor-?1(TGF-?1)-induced epithelial-mesenchymal transition in hepatoma cells SMMC-7721,and to further explore the possible mechanism of its anti-hepatocellular effect.Methods: SMMC-7721 cells were routinely cultured,and SMMC-7721 cells were treated with 10 ng / m L TGF-?1 for 48 hours to prepare human liver cancer SMMC-7721 cells EMT model,Cells were divided into 5 groups:control group,10ng/m L TGF-?1group,low dose group(2?mol/L),medium dose group(4?mol/L)and high dose group(8?mol/L).After 24 hours of intervention with plumbagin,the changes of cell morphology and expression of epithelial-mesenchymal transition markers(E-cadherin,N-cadherin)in the5 treatment groups were detected by immunofluorescence;Cell morphology was observed by Immunofluorescence experiment.The expression of E-cadherin,N-cadherin,Vimentin and Snail were measured at m RNA and protein levelsby RT-PCR and Western blotting,respectively.Western blot was used to detect the expression of PI3 K,AKT,m TOR,p-PI3 K,p-AKT,p-m TOR protein and phosphorylated protein in each group of cells.Results: Plumbagin inhibited TGF-?1-induced cell morphological changes from irregular polygonal to long fusiform.Western blot results showed that plumbagin up-regulated E-cadherin protein expression and down-regulated N-cadherin,Vimentin,Snail protein expression;RT-PCR results showed that plumbagin up-regulated E-cadherin m RNA expression,down-regulated N-cadherin m RNA,Vimentin m RNA,Snail m RNA expression;Western blot showed that after 24 hours of plumbagin treatment,the high concentration plumbagin group can significantly reduce the expression levels of PI3 K,AKT,m TOR,p-PI3 K,p-AKT,p-m TOR protein and phosphorylated protein.After adding PI-103 inhibitor in the treatment,expression levels of PI3 K,AKT,m TOR,p-PI3 K,p-AKT,p-m TOR protein and phosphorylated protein were down-regulated.Conclusions:1.Plumbagin effectively inhibits the epithelial-mesenchymal transition of liver cancer SMMC-7721 cells.2.Plumbagin can significantly reduce the expression levels of PI3 K,AKT,m TOR,p-PI3 K,p-AKT and p-m TOR proteins.The mechanism may be related to the inhibition of PI3 K / Akt / m T0 R signaling pathway.