ABSTRACT
· AIM: To investigate the effect of β1-integrin overexpression on the apoptosis of rabbit corneal epithelial cells and the related mechanism. · METHODS: The plasmid expressing β1-integrin-GFP fusion protein was constructed by polymerase chain reaction (PCR), and this plasmid (β1 group) or the empty vector (mock group) was transfected into rabbit corneal epithelial cells, respectively. The expression of β1-integrin-GFP fusion gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The adhesion of transfected cells to extracellular matrix (ECM) proteins was determined by adhesion assay. The apoptosis of rabbit corneal epithelial cells was assayed by Hoechst 33342 staining and DNA ladder. The phosphorylation of mitogen-activated protein (MAP) kinase was examined by Western blot. · RESULTS: Rabbit corneal epithelial cells overexpressing β1 -integrin-GFP fusion gene were successfully established. Compared with mock group, β1-integrin transfection significantly promoted the adhesive of rabbit corneal epithelial cells to ECM proteins such as laminin, fibronectin, collagen Ⅰ and collagen Ⅳ. Β1-integrin overexpression inhibited apoptosis and induced MAP kinase phosphorylation in rabbit corneal epithelial cells (P<0.05).· CONCLUSION: These data suggest that overexpression of β1-integrin confers resistance to apoptosis in rabbit corneal epithelial cells, and MAP kinase pathway may play an important role in this process.
ABSTRACT
·AIM:To investigate the effect of β1-integrin overexpression on the adhesion and migration of rabbit corneal epithelial (RCE) cells.·METHODS:Eukaryotic expression vector encoding β1-integrin-GFP fusion DNA was transfected into RCE cells,and the β1-integrin-GFP fusion gene was examined by RT-PCR and Western blot.The adhesion to Matrigel and the migration of the transfected cells were determined by adhesion and mobility assays.The phosphorylation of focal adhesion kinase (FAK) was examined by Western blot.·RESULTS:The overexpression of β1-integrin-GFP fusion gene by RCE cells was successfully established.β1-integrin transfection significantly promoted the adhesion of RCE cells to Matrigel ( P < 0.05 ).β1-integrin overexpression also promoted the migration ability of RCE cells and induced FAK phosphorylation in them (P < 0.05).·CONCLUSION:These data suggest that overexpression of β1-integrin promotes the adhesion and migration of RCE cells and that the FAK pathway may play an important role in this process.