Effect of E2F1 overexpression on apoptosis of cerebellar granule neurons

Objective To investigate the effect of E2F1 overexpression on the apoptosis of cerebellar granule neurons(CGNs).Methods The plasmid pE2F1 was constructed and expressed in 293 cells.The transcriptional activity was assessed by 6×E2F-luciferase reporter gene.The CGNs were cultured in vitro and transfected with pE2F1 and pEGFP using calcium-phosphorylation methods.The CGNs were stained with Hoechst 33258 for apoptosis analysis at 12,24,48 h after transfection.Forty-eight hours after transfection with pE2F1,CGNs were subjected to 25 or 5 mmol/L KCl treatment for 12 h and then stained with Hoechst 33258 for apoptosis analysis.Results The pE2F1 was successfully constructed and showed a high level of transcriptional activity.Compared with CGNs transfected with pcDNA at different time points,the apoptotic rate of CGNs transfected with pE2F1 had no significant difference(P0.05).The apoptotic rate of CGNs transfected with pcDNA or pE2F1 in the condition of 25 mmol/L KCl was(12±4)%,(11±3)% respectively,and they had no significant difference.But in the condition of 5 mmol/L KCl,the apoptotic rate of CGNs transfected with pcDNA[(42±3)%] was significantly higher than that of CGNs transfected with pE2F1 [(30±2)%](P0.05).Conclusion Overexpression of E2F1 had no influence on CGNs,but inhibits apoptosis of CGNs induced by 5 mmol/L KCl treatment.