FoxM1 overexpression promotes epithelial-mesenchymal transition and metastasis of hepatocellular carcinoma

Figure 4 Forkhead box protein M1 is required for epithelial-mesenchymal transition. A and B: HepG2 and HUH-7 cells were transfected with 1 ug/mL of pcDNA3.1 or pcDNA3.1-forkhead box protein M1 (FoxM1) for 48 h. The protein/mRNA levels of FoxM1, E-cadherin and vimentin were measured by Western blot and quantitative real-time polymerase chain reaction (qRT-PCR). A densitometric analysis was performed, and the protein/mRNA levels of FoxM1, E-cadherin and vimentin in pcDNA3.1 cells were set to 100% (^aP < 0.05 vs HepG2 pcDNA3.1; ^cP < 0.05 vs HUH-7 pcDNA3.1). Glyceraldehyd-3-phospate dehydrogenase (GAPDH) mRNA was used for normalization; C: HepG2 and HUH-7 cells were transfected with pcDNA3.1 or pcDNA3.1-FoxM1 for 48 h, and the typical morphology changes were measured using a light microscope; D: HepG2 and HUH-7 cells were transfected with pcDNA3.1 or pcDNA3.1-FoxM1 for 48 h, and the invasion changes was measured by Transwell assay (^aP < 0.05 vs pcDNA3.1); E: The HepG2 and Huh7 cells were transfected with 100 nmol/L of each siRNA for 48 h. The protein/mRNA levels of FoxM1 were measured by Western blot and qRT-PCR. A densitometric analysis was performed, and the protein/mRNA levels of FoxM1 in control-siRNA cells were set to 100% (^aP < 0.05 vs control-siRNA; ^cP < 0.05 vs FoxM1-siRNA-1). GAPDH mRNA was used for normalization. Mixed siRNA was used in subsequent experiments; F and G: HepG2 and HUH-7 cells were transfected with FoxM1-siRNA or control-siRNA in the presence of HGF for 48 h, and the protein/mRNA levels of FoxM1, E-cadherin and vimentin were measured by Western blot and qRT-PCR. A densitometric analysis was performed, and the protein/mRNA levels of FoxM1, E-cadherin and vimentin in control-siRNA cells were set to 100% (^aP < 0.05 vs HepG2 control-siRNA; ^cP < 0.05 vs HUH-7 control-siRNA). GAPDH mRNA was used for normalization; H: HepG2 and HUH-7 cells were transfected with FoxM1-siRNA or control-siRNA in the presence of hepatocyte growth factor (HGF) for 48 h, and the invasion changes was measured by Transwell assay (^aP < 0.05 vs control-siRNA).