3-MA对三氧化二砷诱导Jurkat细胞凋亡作用的影响  

Effect of 3-MA on arsenic trioxide induced apoptosis in Jurkat cells

作　　者：王艳婕[1] 牛志国[1] 郭继强[1] 王辉[1] 牛新清[1] 

机构地区：[1]新乡医学院免疫学研究中心,新乡453003

出　　处：《中国免疫学杂志》2014年第11期1472-1476,共5页Chinese Journal of Immunology

基　　金：河南省卫生厅资助计划(No.2011020098)

摘　　要：目的:探讨自噬特异性抑制剂3-甲基腺嘌呤(3-MA)对三氧化二砷(As2O3)诱导急性T细胞白血病系Jurkat细胞系Jurkat细胞凋亡的影响及机制。方法:XTT法检测三氧化二砷(As2O3)对急性T细胞白血病细胞系Jurkat细胞的增殖抑制作用。电镜下观察不同浓度As2O3作用Jurkat细胞24 h后细胞形态。免疫印迹和流式细胞术检测微管相关蛋白1轻链3B(LC-3B)蛋白的表达变化。AnnexinV-FITC/PI双染法流式细胞术检测3-MA对AS2O3诱导急性T细胞白血病系Jurkat细胞凋亡的影响。结果:As2O3可以抑制急性T淋巴细胞白血病细胞株Jurkat细胞的生长,这种作用呈剂量和时间依赖性。2.5、5、10μmol/L AS2O3作用Jurkat细胞24 h后在电镜下可观察到自噬、凋亡、坏死的不同形态,并且自噬体数目不断增多。5μmol/L As2O3处理Jurkat细胞0、24、48 h后,LC-3B的平均荧光强度相对倍数分别(3.1±0.2)倍、(4.6±0.31)倍、(34.2±4.5)倍,组间相比,差异有统计学意义(P<0.05),与生长抑制率一样呈现时间依赖性增强;免疫印迹也显示As2O3处理24 h和48 h后,LC-3B的蛋白表达逐渐增强;相对于As2O3组(33.4±9.1)%的生长抑制率,3-甲基腺嘌呤(3-MA)联合As2O3处理细胞后,48 h的生长抑制率为(60.6±8.3)%,差异明显,而LC-3B的表达明显降低。联合应用自噬抑制剂3-MA后Jurkat细胞的凋亡率(44.96±3.60)%,与单用As2O3组(2.94±0.26)%相比明显增加,差异有统计学意义。结论:自噬抑制剂3-MA可以增加As2O3引起的急性T细胞白血病细胞系Jurkat细胞的凋亡细胞百分数,其作用机制与诱导凋亡及抑制自噬密切相关。Objective： To discuss the effect and mechanism of autophagy inhibitor 3-MA on arsenic trioxide inducing apoptosis of acute T-cell leukemia cell line Jurkat cells. Methods： Proliferation inhibition of Jurkat cells treated with arsenic trioxide was detected by XTT. Morphological characteristics of Jurkat cells treated with different concentrations arsenic trioxide were observed by electron microscope. Microtubule-associated protein 1 light chain 3B（ LC-3B） protein expression was detected by Western blot and flow cytometry. Apoptosis rates of Jurkat cells treated with 3-MA combining arsenic trioxide were detected by flow cytometry using AnnexinV-FITC /PI double staining. Results： Arsenic trioxide inhibited the growth of Jurkat cells in a dose and time dependence. We observed different morphological characteristics of autophagy,apoptosis and necrosis accompanying more autophagosomes in Jurkat cells which were treated with arsenic trioxide 2. 5,5,10 μmol /L after 24 h. LC3 B mean fluorescence intensity（ MFI） relative multiples were（ 3. 1 ± 0. 2）fold,（ 4. 6 ± 0. 31） fold,（ 34. 2 ± 4. 5） fold with 5 μmol /L arsenic trioxide treated Jurkat cells 0,24,48 h,and the P values between each of the two groups were less than 0. 05,which increased depending time consistently with the growth inhibition rates. LC-3B protein expression gradually increased treated Jurkat cells with arsenic trioxide after 24 h,48 h. The growth inhibition rate（ 60. 6 ± 8. 3） % was significantly different treated with arsenic trioxide combining 3-methyl adenine（ 3-MA） while it was（ 33. 4 ± 9. 1） % treated with arsenic trioxide alone,however,LC-3B protein expression gradually decreased. Jurkat cell apoptosis rate（ 44. 96 ± 3. 60） % was significantly increased treated with arsenic trioxide combining autophagy inhibitor（ 3-MA） while it was（ 2. 94 ± 0. 26） % treated with arsenic trioxide alone,and this difference was statistically significant. Conclusion： 3-MA increased apoptosis rates of Jurkat c

关 键 词：三氧化二砷 自噬 凋亡 急性T细胞白血病 

分 类 号：R733.71[医药卫生—肿瘤]