Figure 5.
PFKFB3 regulated the expression of EMT-related molecules in GC cells. Real-time RT-PCR and western blot analyses revealed PFKFB3 overexpression significantly promoted the expression of N-cadherin, Vimentin, Snail, and Twist, inhibited E-cadherin at both mRNA and protein levels in SGC-7901 cells, whereas the expression of Zeb1 did not show obvious changes (a and b). All data are shown as mean ± SD, ***P < 0.001 (a). SGC-7901 cells with stable PFKFB3 overexpression were pretreated with 1-mM helenalin for 60 minutes. Western blot analysis showed that helenalin pretreatment could suppressed PFKFB3-induced the expression of EMT-related molecules in SGC-7901 cells (b). RT-PCR and western blot analyses revealed PFKFB3 knockdown expression significantly inhibited the expression of N-cadherin, Vimentin, Snail, and Twist, promoted E-cadherin at both mRNA and protein levels in SGC-7901 cells, whereas the expression of Zeb1 did not show obvious changes (c and d). All data are shown as mean ± SD, **P < 0.01; ***P < 0.001 (c). AGS cells with PFKFB3 shRNA were incubated with 100-ng/mL NF-κB p65 for 24 hours, or AGS cells with stable PFKFB3 knockdown expression were transfected with NF-κB p65–overexpressing lentivirus. Western blot analysis showed that NF-κB p65 treatment could rescue PFKFB3 knockdown expression-mediated the expression of EMT-related molecules in AGS cells (d). EMT, epithelial-mesenchymal transition; RT-PCR, reverse transcription polymerase chain reaction.