Differences In Gene Expression Related To Epithelial-Mesenchymal Transition In Patients With Liver Cancer And The Mechanism Of Qizhu Recipe

Objectives1.To reveal the role of epithelial-mesenchymal transition(EMT)in the development and progression of primary liver cancer by comparing the expression differences of genes related to EMT between patients and healthy controls and,at the same time,between different BCLC stages;2.To study in vitro whether Qizhu recipe has inhibitory effect on TGF-β1 induced EMT,so as to provide material basis for its clinical application.Methods1.PBMC in peripheral blood of liver cancer patients and healthy control group was extracted,gene expression was obtained by high-throughput sequencing,EMT-related gene data was screened,and differences between the two groups were analyzed;In the liver cancer group,differences in EMT-related gene expression were compared according to BCLC stages,and the correlation between clinical biochemistry and EMT genes was explored by Spareman;2.Using TGF-β1 to induce EMT in HepG2 cells in vitro,and the HepG2 cells were co-cultured with Qizhu recipe.Cell morphology was observed under the microscope,cell migration was observed in the scratch experiment,and the expression of EMT-related proteins was detected by western blot.Results1.The expressions of TGFB1,CDH1、CDH2,CTNNB1,VIM,SNAI1,TWIST2,ZEB1 and ZEB2 in the liver cancer group were all higher than those in the healthy control group(all P<0.05);2.Liver cancer patients were divided into groups 0~B and C/D according to BCLC stages.There were no statistically significant differences in CDH2(P>0.05),while the expressions of VIM,SNAI1 and ZEB1 in C/D group were lower than those in 0~B group(all P<0.05).3.In Spearman correlation analysis,the correlation coefficients of GGT,TGFB1,CDH2 and ZEB1 were-0.401,-0.455 and-0.450,respectively,showing a negative correlation with statistical significance(P<0.05).The correlation coefficient between ALB and SNAI1 was 0.365,with positive correlation and statistical significance(P<0.05).SNAI1,ZEB1 and ZEB2 were positively correlated with mesenchymal phenotypic genes CDH2,VIM and CTNNB1,with statistical significance(all P<0.01).The correlation coefficients of ZEB2 and VIM and CTNNB1 were 0.979 and 0.940,while the correlation coefficients of ZEB1 and CTNNB1 were 0.940.4.Qizhu recipe can inhibit the TGF-β1 induced HepG2 cell morphological changes of EMT and reduce cell migration.The scratch experiment shows that compared with the TGF-β1 group,difference was statistically significant(P<0.05).After ading TGF-β1 in HepG2 cells,WB shows that E-cadherin expression reduced,while N-cadherin,Vimentin,Snail and Fibronectin increased.Among them,Fibronectin and the control group have statistically significant difference(P<0.05).Then in the groups added Qizhu recipe,E-cadherin expression increased in concentration dependence.Comparing with the control group,the difference was statistically significant(P<0.05).N-cadherin,Vimentin,Snail and Fibronectin expression decreased,Comparing with TGF-β1 group,Vimentin,Fibronectin and Snail had statistical significance(P<0.05).Conclusions1.The EMT program was relatively activated in liver cancer patients.In patients with C/D stage,the mesenchymal phenotypic markers were relatively reduced,showing MET reversal.2.EMT-TFs plays an important role in the expression of mesenchymal phenotypic genes,among which the ZEB family may be regulated by Wnt/β-catenin pathway.3.Qizhu recipe can inhibit the EMT changes of HepG2 cells induced by TGFβ1,and reduce migration.