BACKGROUND: Epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) contributes to posterior capsule opacification (PCO). C-terminal binding protein 2 (CtBP2) has been reported to be essential in EMT and embryonic development. However, the function of CtBP2 in EMT of LECs is unknown. The goal of this study was to investigate the role of CtBP2 through Notch signaling in transforming growth factor β2 (TGFβ2)-induced EMT in LECs. METHODS: The human LEC line SRA01/04 was cultured in the presence of TGFβ2 for different periods of time or with γ-Secretase Inhibitor IX (DAPT), a specific inhibitor of Notch receptor cleavage, for 24 h, utilizing plasmid-based method. The levels of protein expression of CtBP2, EMT markers, and Notch signaling molecules were measured by Western bolts. RESULTS: Treatment of SRA01/04 cells with TGFβ2 induced typical molecular changes of EMT and increased the expression of CtBP2 in a time-dependent manner. Similarly, the expressions of Jagged1 and Notch1 were increased after TGFβ2 treatment. Knockdown of CtBP2 by specific siRNA inhibited TGFβ2-induced changes of Connexin 43 (CX43), α-smooth muscle actin (α-SMA), Notch1, and Notch intracellular domain (NICD). In addition, treatment of LECs with ectopic expression of CtBP2 changed the expressions of CX43, α-SMA, Notch1, and NICD, but blockade of Notch pathway with DAPT inhibited CtBP2-induced changes of α-SMA and CX43. CONCLUSION: Our data suggest that CtBP2 plays a critical role in TGFβ2-induced EMT via the Jagged/Notch signaling pathway in human LECs and may contribute to the development of PCO.
BACKGROUND: Epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) contributes to posterior capsule opacification (PCO). C-terminal binding protein 2 (CtBP2) has been reported to be essential in EMT and embryonic development. However, the function of CtBP2 in EMT of LECs is unknown. The goal of this study was to investigate the role of CtBP2 through Notch signaling in transforming growth factor β2 (TGFβ2)-induced EMT in LECs. METHODS: The humanLEC line SRA01/04 was cultured in the presence of TGFβ2 for different periods of time or with γ-Secretase Inhibitor IX (DAPT), a specific inhibitor of Notch receptor cleavage, for 24 h, utilizing plasmid-based method. The levels of protein expression of CtBP2, EMT markers, and Notch signaling molecules were measured by Western bolts. RESULTS: Treatment of SRA01/04 cells with TGFβ2 induced typical molecular changes of EMT and increased the expression of CtBP2 in a time-dependent manner. Similarly, the expressions of Jagged1 and Notch1 were increased after TGFβ2 treatment. Knockdown of CtBP2 by specific siRNA inhibited TGFβ2-induced changes of Connexin 43 (CX43), α-smooth muscle actin (α-SMA), Notch1, and Notch intracellular domain (NICD). In addition, treatment of LECs with ectopic expression of CtBP2 changed the expressions of CX43, α-SMA, Notch1, and NICD, but blockade of Notch pathway with DAPT inhibited CtBP2-induced changes of α-SMA and CX43. CONCLUSION: Our data suggest that CtBP2 plays a critical role in TGFβ2-induced EMT via the Jagged/Notch signaling pathway in human LECs and may contribute to the development of PCO.