Aim: The relationship between latent adenovirus infection and apoptosis of airway epithelial cell have not been well documented.We want to illustrating the roles of adenovirus E1A protein on the apoptotic alveolar epithelial in response to TNF-alpha.
Methods: The expression vector for expressing adenovirus E1A protein was transfected into CCL149 and A549 cell respectively. Cell stably expressing E1A protein were selected by G418 resistance.All G418-resistant clones were indentified by RT-PCR and immunocytochemistry. The rate of apoptosis were measured by Hoeschest 33 258 and flow cytometry respectively. The apoptotic rate in response to 30 microg/L TNF-alpha was compared between E1A-positive clones and control clones both in A549 and CCL149.
Results: The rates of apoptosis were (2.63+/-0.8)%, (25.38+/-0.9)% respectively in E1A-positive CCL149 cell and (0.62+/- 0.3)%, (6.08+/-0.2)% respectively in E1A-negative CCL149 cell. The rates of apoptosis were (2.63+/-0.8)%, (25.38+/-0.9)% respectively in E1A-positive A549 cell and (0.62+/- 0.3)%, (6.08+/-0.2)% respectively in E1A-negative A549 cell. The rate of apoptosis were increased in E1A-positive cells compared with control with or without TNF-alpha stimulation.
Conclusion: E1A sensitizes cell to TNF-alpha induced apoptosis of A549 cell and CCL149 cell.