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.
出处
《基础医学与临床》
CSCD
北大核心
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)
