SARI基因促进急性髓性白血病细胞凋亡机制的探讨  

Mechanism of SARI gene promoting apoptosis in acute myeloid leukemia cells

作　　者：薛龑[1] 陈梅环 唐永金 吴玮 王小花 徐建萍[1] 林东红[1] 

机构地区：[1]福建医科大学医学技术与工程学院,福州350004 [2]福建省妇幼保健院,福州350001

出　　处：《中国免疫学杂志》2017年第11期1621-1625,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(81371879)资助

摘　　要：目的:通过慢病毒载体过表达SARI(Suppressor of AP-1,regulated by IFN)基因,探讨SARI基因促进急性髓性白血病(Acute myeloid leukemia,AML)细胞凋亡的分子机制。方法:构建SARI基因过表达的慢病毒载体,感染AML细胞HL-60和NB4,应用实时荧光定量PCR和Western blot对感染后细胞进行过表达的鉴定。两株AML细胞均设空白对照组(CON组)、空载体对照组(NC组)及SARI组,应用Western blot检测SARI过表达后两株AML细胞组凋亡相关蛋白Bcl-2、Bcl-xl、Bax和凋亡途径相关蛋白CytoC、Caspase8、Caspase9、Caspase3、Actived-Caspase3、PARP的表达变化。结果:SARI组细胞的SARI mRNA和蛋白表达水平均显著高于CON组和NC组(P<0.05),表明SARI过表达成功。SARI过表达的HL-60和NB4细胞中抗凋亡蛋白Bcl-2、Bcl-xl表达均下调,促凋亡蛋白Bax表达上调。CytoC表达增加,Caspase8、Caspase9、Caspase3剪切激活,Actived-Caspase3上调,PARP下调,PARP剪切体上调。结论:SARI基因促进AML细胞凋亡可能是通过激活外源性凋亡途径及与外源性凋亡途径交叉的内源性凋亡途径。Objective：To explore the apoptotic mechanisms of acute myeloid leukemia（AML） cells by a lentivirus mediatedSARI（suppressor of AP-1,regulated by IFN） overexpression system.Methods：The lentiviral vectors of overexpression SARI were transfected into AML cell HL-60 and NB4.Real-time quantitative PCR and Western blot were employed in the following evaluation tests.Both the cells were all divided into blank control group（CON group）,empty vector control group（NC group） and SARI overexpression group（SARI group）.Western blot was used to detect the expressions of apoptosis-related proteins Bcl-2,Bcl-xl,Bax and apoptotic pathway related proteins CytoC,Caspase8,Caspase9,Caspase3,Actived-Caspase3,PARP.Results：The SARI groups exhibited a high level of SARI mRNA and SARI protein when compared with CON group and NC group（P0.05）.In both the SARI groups,the expression of anti-apoptotic proteins Bcl-2,Bcl-xl were down-regulated and the pro-apoptotic protein Bax was up-regulated.CytoC increased,Caspase8,Caspase9 and Caspase3 decreased,Actived-Caspase3 up-regulated.PARP down-regulated,and PARP spliced variant up-regulated.Conclusion：SARI might promote the apoptosis of AML cells through activing the exogenous and endogenous apoptotic pathways which crossed to exogenous.

关 键 词：SARI AML 细胞凋亡 凋亡途径 分子机制 

分 类 号：R733.72[医药卫生—肿瘤]