Overexpression of Bax and induction of apoptosis in human retinal pigment epithelial cell lines

Purpose To investigate bax expression and induction of apoptosis in normal cultured retinal pigment epithelium (RPE) cells . [WT5”HZ]Methods [WT5”BZ]Cultured human RPE cells were transfected by P MDNA3 hbax ,which incoded the whole bax gene and may be induced by Zn 2+ under the MTII promoter, through lepofectin mediated protocol.The tested RPE cells were divided into three groups of A, P MDNA3 hbax transfected ;B,P MDNA3 (nude vector) transfected and C,normal RPE cells.After transfection, DNA gel electrophoreses were performed ,the tested RPE cell cycles were analyzed with flow cytometry (FCM). Results The gel electrophoretogram showed DNA ladder phenomenon,FCM confirmed the apoptosis of RPE cells P MDNA3 bax transfected , consisting of significant apoptotic peak sited before the G 1 phase and the apoptotic rate was 36%. Conclusion The foreign bax gene can be effectively conducted into the RPE cell through lepofectin mediated protocol and induced expression . The foreign bax overexpression may induce the cultured human RPE cell susceptibility to apoptosis.