稳定表达反义ATMPI-3K的细胞系U937-ASPI-3K细胞凋亡敏感性  

The sensitivity to apoptosis is enhanced in U937-ASPI-3K cells which stably express antisense ATM/PI-3K

作　　者：周剑锋[1] 刘文励[1] 戴琪琳 孙岚[1] 汤屹[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院,武汉430030

出　　处：《中华血液学杂志》2000年第12期641-643,共3页Chinese Journal of Hematology

基　　金：国家自然科学基金!资助项目 (3980 0 14 9)

摘　　要：目的　分析稳定表达反义ATM基因细胞系U937 ASPI 3K的细胞凋亡敏感性特征。方法　用膜联蛋白Ⅴ 碘化丙锭凋亡检测试剂盒和流式细胞仪分析凋亡率。用Westernblot分析细胞色素C和抗凋亡蛋白Bcl 2表达水平。结果　U937 ASPI 3K细胞凋亡敏感性显著增强 ,1.5Gy1 3 7Cs照射后 8,48h细胞凋亡率分别为 2 8.0 %和 (5 3 .0± 5 .4) % ,对照组U937 pZEOSV2 (+ )分别为 4.2 %和 (11.0±3 3) %。受照后 4～ 8h ,U937 ASPI 3K细胞内出现逐渐增强的细胞色素C蛋白条带 ,对照组未检测到相应的蛋白条带。受照后 4～ 8h ,U937 ASPI 3K细胞Bcl 2蛋白水平逐渐降低 ,对照组Bcl 2蛋白水平无明显变化。结论　稳定表达反义ATM基因的细胞系U937 ASPI 3K继发性获得凋亡敏感性增强表型 ,说明ATM基因的选择灭活在肿瘤治疗中有重要意义。Objective To analyse the characteristics of apoptosis sensitivity in U937 ASPI 3K cells which is stably expressing anti sense ATM/PI 3K. Method Annexin Ⅴ PI apoptosis detection kid and flow cytometry were used to determine apoptosis rate, Western blot to the expression level of cytochrome C protein and Bcl 2 protein . Results U937 ASPI 3K cells had a lowered threshold for triggering apoptosis in response to low dose irradiation. Cell apoptosis rate was 28% and (53.0±5.4)%, respectively, at 8 and 48 hours after 1.5?Gy 137 Cs irradiation, while in U937 pZEOSV2(+) control group it was 4.2% and (11.0±3.3)%, respectively. Cytosolic cytochrome C protein band was gradually intensified in U937 ASPI 3K in 4～8h after irradiation, but was not detected in control group. Bcl 2 protein gradually decreased in U937 ASPI 3K in 4～8?h after irradiation, but didn't change obviously in control group. Conclusion Cell line U937 ASPI 3K acquired a phenotype with a lowered threshold for triggering apoptosis. It demonstrated that selective inactivation of ATM gene might be of considerable value in tumor treatment.

关 键 词：反义寡核苷酸 U937-ASPI-3K 细胞凋亡 治疗 肿瘤 

分 类 号：R346[医药卫生—基础医学]