PTD-OD-HA融合蛋白对bcr/abl阳性细胞凋亡的影响  

The influence of PTD-OD-HA fusion proteins on apoptosis of bcr/abl positive cell lines

作　　者：黄峥兰[1] 季茂胜[1] 袁颖[2] 黄世峰[1] 刘钉宾[1] 曾建明[1] 温健萍 冯文莉[1] 

机构地区：[1]重庆医科大学临床血液学教研室,临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附属第一医院普外科,重庆400016 [3]Canadian Blood Services, Hamilton ON L8S 1 H8 ,Canada

出　　处：《肿瘤》2010年第4期267-271,共5页Tumor

基　　金：国家自然科学基金资助项目(编号:30670901)

摘　　要：目的:研究含有蛋白转导域(protein transduction domain,PTD)、寡聚化结构域(oligomerization domain,OD)和HA标签的PTD-OD-HA融合蛋白对几种bcr/abl阳性细胞株细胞凋亡的影响。方法:将前期制备的PTD-OD-HA蛋白作用于几种bcr/abl阳性细胞后,应用FCM法、DNA梯度电泳(DNAladder)和透射电子显微镜检测细胞凋亡,RT-PCR和Western印迹法检测凋亡相关基因bax、bcl-2的mRNA和蛋白水平变化。结果:FCM法检测发现,PTD-OD-HA能促进bcr/abl阳性细胞发生凋亡;DNA ladder实验检测发现,PTD-OD-HA作用后BaF3-P210和K562细胞DNA电泳有梯状条带;透射电子显微镜观察发现,PTD-OD-HA作用后BaF3-P210细胞发生了凋亡;RT-PCR及Western印迹法检测显示,促凋亡基因bax的mRNA及蛋白水平增高,抗凋亡基因bcl-2的mRNA及蛋白水平降低。结论:PTD-OD-HA蛋白可以特异性诱发bcr/abl阳性细胞的凋亡。Objective：To study the influence of protein transduction domain（PTD）-oligomerization domain（OD）-HA fusion proteins on apoptosis of bcr/abl-positive cell lines.Methods：bcr/abl-positive cells were treated with PTD-OD-HA protein.The apoptoses of the cells were detected by flow cytometry（FCM）,DNA ladder and transmission electron microscopy（TEM）,and the levels of apoptosis-related genes bax and bcl-2 were detected by RT-PCR and Western blotting.Results：FCM examination demonstrated that PTD-OD-HA protein induced the apoptosis of bcr/abl-positive cells;DNA ladder showed that the classic DNA ladders appeared in BaF3-P210 and K562 cells after 48 h treatment with PTD-OD-HA proteins;the apoptoses of BaF3-P210 cells were observed by TEM;the levels of bax in mRNA and protein increased in BaF3-P210 and K562 cells,and bcl-2 decreased.Conclusion：PTD-OD-HA proteins specifically induced the apoptosis of bcr/abl positive cells.

关 键 词：白血病 髓样 慢性 重组融合蛋白质类 细胞凋亡 融合蛋白类 bcr-abl 

分 类 号：R730.54[医药卫生—肿瘤]