摘要
目的 研究外源性促凋亡基因 bax过表达对培养人的视网膜色素上皮 (retinal pigment ep-ithelium,RPE)细胞凋亡的影响。 方法 通过脂质体 L ipofectin介导法用含人全长 bax基因片段的真核表达载体 PMDNA3- hbax质粒转化体外培养的 RPE细胞 ,用流式细胞仪检测细胞周期的变化 ,提取实验组细胞DNA进行 DNA梯度电泳。 结果 Zn2 + 可诱导 PMDNA3- hbax转化组 RPE细胞出现明显的凋亡峰 ,凋亡峰位于单倍体和二倍体之间 ,凋亡率为 36 % ,DNA电泳可见 DNA梯状条带 ,PMDNA3空载体组及未转染组 RPE细胞未见亚二倍峰和 DNA梯状条带出现。 结论 外源性促凋亡基因 bax过表达可诱导 RPE细胞凋亡。
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.
出处
《中华眼底病杂志》
CAS
CSCD
2001年第2期132-134,共3页
Chinese Journal of Ocular Fundus Diseases
基金
国家自然科学基金!资助项目 (3970 0 1 54)
