Effects of BTg-3 on proliferation,invasion,migration and Akt/GSK-3β/β-catenin signaling pathway of liver cancer cells
Objective The aim of this study was to investigate the effect of B cell translocation gene 3(BTg-3)on prolifera-tion,invasion,migration and protein kinase B/glycogen synthase kinase-3β/β-catenin(Akt/GSK-3β/β-catenin)signaling pathway of liver cancer cells.Methods Western blot was used to measure the expression of BTg-3 protein in liver cancer BEL-7402 cells,SMMC-7721 cells,HepG2 cells and normal liver cancer LO2 cells.HepG2 cells were divided into the blank control group(N group),empty vector group(V group),BTg-3 overexpression group(BTg-3 group)and BTg-3 overexpression+LICL group(BTg-3+LICL group).HepG2 cells in the N group were not transfected and HepG2 cells of the V group were transfected with empty vector pcDNA 3.1.HepG2 cells in the BTg-3 group were transfected overexpressing BTg-3 vector pcDNA3.1-BTg-3,while HepG2 cells in the BTg-3 overexpression+LiCl group were transfected with the BTg-3 vector pcDNA3.1-BTg-3 and treated with 10 nmol/l GSK-3β inhibitor-lithium chloride(LiCl).MTT assay was used to determine the proliferative ability of HepG2 cells in each group.The cell colony formation assay was used to determine the survival ability of HepG2 cells in each group.Transwell assay was used to determine the invasive and migratory ability of HepG2 cells in each group.Western blot was used to determine the expression of Akt,phosphoryl-ated Akt(p-Akt),p-GSK-3β,β-catenin,Cyclin D1,N-cadherin and epithelial cadherin(E-cadherin)in HepG2 cells.Results The expression of BTg-3 protein in human liver cancer BEL-7402 cells,SMMC-7721 cells and HepG2 cells were lower than that in normal liver LO2 cells(P<0.001).Compared with the N group and V group,the A values and clone formation rates were decreased in the BTg-3 group and BTg-3+LICL group(P<0.05),as well as a decrease in the number of invasive cells and migratory cells(P<0.05).The expression of p-Akt,p-GSK-3β,β-catenin,Cyclin D1 and N-cadherin proteins decreased(P<0.05),and the expres-sion of BTg-3 and E-cadherin proteins increased(P<0.05)in the BTg-3 group and BTg-3+LICL group.Compared with the BTg-3 group,the A values and clone formation rate in the BTg-3+LICL group increased(P<0.05),as well as an increase in the number of invasive cells and migratory cells(P<0.05).The expression of p-Akt,p-GSK-3β,β-catenin,Cyclin D1 and N-cadherin proteins increased(P<0.05),while the expression of BTg-3 and E-cadherin proteins in HepG2 cells decreased in the BTg-3+LICL group(P<0.05).There were no significant differences in the indexes of HepG2 cells between the N group and V group(P>0.05).Conclusion BTg-3 is lowly expressed in liver cancer cells,and overexpression BTg-3 may inhibit proliferation,invasion and migration of liver cancer cells through the Akt/GSK-3β/β-catenin signaling pathway.
B cell translocation gene 3Liver cancerEpithelial-mesenchymal transition
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