过表达突变的NPM1基因对THP-1白血病细胞增殖和凋亡的影响  

Effects of overexpression nucleophosmin(NPM1) mutations on the proliferation and apoptosis of THP-1 leukemic cells

作　　者：覃凤娴[1] 邵会媛[1] 陈先春[1] 谭诗[1] 张慧娟[1] 肖玲[2] 张伶[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室重庆市重点实验室,重庆400016 [2]重庆医科大学附属儿童医院检验科,重庆400014

出　　处：《基础医学与临床》2012年第3期251-256,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30872418);重庆市科委自然科学基金(CSTC2010BB5363)

摘　　要：目的探讨核仁磷酸蛋白基因(NPM1)突变对白血病细胞增殖和凋亡的影响。方法将携带NPM1 A型突变(NPM1 mA)的重组质粒载体pEGFPC1-NPM1 mA转染白血病THP-1细胞系,构建稳定表达NPM1 mA蛋白的细胞株(THP-1 mA),同时设立空载体转染组(THP-1 C1)和未处理组(THP-1)为对照。MTT实验观察细胞增殖;流式细胞术分析细胞周期和凋亡;RT-PCR和Western blot分别检测细胞凋亡相关蛋白(BAX和BCL-2)mRNA及蛋白表达水平。结果与对照组相比较,实验组THP-1 mA细胞体外增殖能力明显增强(P<0.05),S期细胞比例明显增高(P<0.05),G1期细胞比例显著减低(P<0.05);而3组细胞凋亡率无显著差异,BAX,BCL-2的mRNA和蛋白表达以及BAX/BCL-2比值亦未见明显改变。结论 NPM1突变基因能够促进白血病细胞的体外增殖能力,而对白血病细胞凋亡无显著影响。Objective To investigate the effect of Nucleophosmin（NPM1） mutations on the proliferation and apoptosis of leukemia cells.Methods The pEGFPC1-NPM1 mA plasmid vector was transfected into THP-1 cells to establish the stably expressed NPM1 mutation A protein leukemia cells（THP-1 mA）.The cells transfected with pEGFPC1 plasmid（THP-1 C1） and the untreated cells（THP-1） were set as control.Cells proliferation potential was assessed by MTT assay;flow cytometry was used to detect the cell cycle and cellular apoptosis.The mRNA and protein expression of BAX and BCL-2 were analyzed by RT-PCR and Western blot,respectively.Results Compared with the controls,growth ability of THP-1 mA cells was significantly improved（P0.05）.Additionally,the percentage of cells in the S phase increased significantly and that in the G_1 remarkably decreased（P0.05）.While there was no significantly change on cellular apoptosis and the expression of BAX and BCL-2 on the mRNAlevel or protein level.The ratio of BAX/BCL-2 also had no significantly change.Conclusions NPM1 mutations may promote cells proliferation potential in vitro,but had no significantly influence on cellular apoptosis.

关 键 词：NPM1突变 白血病 THP-1细胞系 增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]