Effects of Shangxian granules on migration and invasion of Hepa1-6 hepatocellular carcinoma cells and Wnt/β-catenin signaling pathway
Objective It is to observe the effects of serum containing Shanxian granules on migration and invasiveness of hepatocellular carcinoma Hepa1-6 cell line,and explore its possible molecular mechanisms based on epithelial-mesen-chymal transition(EMT)and Wnt/β-catenin signaling pathways.Methods The SD rats were used to prepare Shanxian granule low,medium and high dose drug-containing serum and blank serum.Hepa1-6 hepatocellular carcinoma cells were cultured in vitro,and divided into blank serum group,KYA1797K group and Shanxian granules low,medium and high dose drug-containing serum groups.Hepa1-6 hepatocellular carcinoma cells were taken from each group after 12 h,24 h and 36 h of intervention,and their migration ability was detected by scratch assay,invasion ability by Transwell assay,and the protein expressions of E-cadherin,N-cadherin,vimentin,β-cadherin and phosphorylated glycogen synthase kinase 3β(p-GSK3β)in the cells were detected by Western blotting.Results Compared with the blank serum group,the cell migra-tion distances after 12 hours of intervention in the medium and high-dose medicated serum groups of Shanxian granule and KYA1797K group,and the cell migration distance after 24 and 36 hours of intervention in every Shanxian granule group and KYA1797K group was also significantly reduced(all P<0.05).After12 h,24 h,and36 h of intervention,the numbers of invading cells in each group of Shanxian granule and KYA1797K group were significantly reduced(all P<0.05),while the relative expression of E-cadherin protein in the cells was significantly increased(all P<0.05),additionally,the relative expressions of N-cadherin,Vimentin,β-catenin,and p-GSK3β proteins in the cells were significantly decreased(all P<0.05).The cell migration distance at each time point in the low,medium,and high-dose medicated serum groups of Shanxian granule showed a dose-dependent reduction,the number of invading cells displayed a dose-dependent decrease,the relative expressions of E-cadherin,N-cadherin,and Vimentin proteins in the cells exhibited a dose-dependent increase or decrease,the differences between the groups were all statistically significant(all P<0.05).Conclusion Shanxian gran-ule exhibited a dose-dependent inhibition on the migration and invasion of Hepa1-6 hepatocellular carcinoma cells.This effect may be attributed to its ability to upregulate E-cadherin expression and downregulate the expression of N-cadherin and Vimentin through the Wnt/β-catenin pathway,ultimately inhibiting the epithelial-mesenchymal transition(EMT)process in Hepa1-6 hepatocellular carcinoma cells.
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