The influence of HNF4G on the proliferation and migration of gallbladder cancer cells and the underlying mechanism / 肿瘤

ABSTRACT

Objective:To investigate the effect of hepatocyte nuclear factor 4 gamma(HNF4G)on the proliferation,migration,cell cycle and apoptosis of gallbladder cancer cells and the underlying molecular mechanisms. Methods:The expression levels of HNF4G mRNA and protein in gallbladder cancer cell lines(GBC-SD,NOZ and ZJU-0430)were examined by real-time fluorescence quantitative PCR and Western blotting,respectively.Stable HNF4G-silencing or overexpressing GBC-SD,NOZ and ZJU-0430 cell lines were established by lentiviral infection.Then,the effect of HNF4G silencing or overexpression on the proliferation,migration,cell cycle and apoptosis of gallbladder cancer cell lines were evaluated by CCK-8 assay,colony formation assay,Transwell assay and FCM assay.The effect of changes in HNF4G gene expression level on the expression levels of cell cycle associated proteins(Cyclin A2 and Cyclin B1),apoptotic proteins(Bax and Bcl-2),epithelial-mescenchymal transition associated proteins(E-cadherin,N-cadherin,vimentin,Snail and Slug)as well as ErbB2 and phosphorylated AKT in the ErbB2/PI3K/AKT signaling pathway was examined by Western blotting. Results:Real-time fluorescence quantitative PCR and Western blotting results demonstrated that the expression level of HNF4G mRNA and protein in NOZ cells was significantly lower than that in GBC-SD and ZJU-0430 cells(P＜0.001).The successful establishment of HNF4G-sliencing and HNF4G-overexpressing gallbladder cancer cell lines was verified by Western blotting analysis.Over expression of HNF4G could enhance the proliferation and migration of GBC-SD,NOZ and ZJU-0430 cells(P＜0.001),promote the transition of S-phase to G2/M-phase,and inhibit the apoptosis of these gallbladder cancer cell lines.Western blotting analysis showed that overexpression of HNF4G could increase the expression of Cyclin A2,Cyclin B1,Bcl-2,N-cadherin,Vimentin,Snail,Slug,ErbB2 and phosphorylated AKT,and decrease the expression of Bax and E-cadherin(P＜0.001).In contrast,HNF4G silencing could induce the opposite changes in the biological behaviors and protein expression in GBC-SD cells. Conclusion:HNF4G affects the proliferation and migration of gallbladder cancer cells by participating in the regulation of the epithelial-mescenchymal transition and ErbB2/PI3K/AKT signaling pathway.The influence of HNF4G on gallbladder cancer cells suggests that HNF4G may be a potential gene target for gallbladder cancer treatment.