NPM1基因表达对急性髓系白血病细胞系的影响及其机制探讨  

Effects of NPM1 gene expression on acute myeloid leukemia cell lines and its mechanism

作　　者：邱少伟 万玉玲 王敏[1] 王建祥 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院,天津300020

出　　处：《中华血液学杂志》2017年第11期940-944,共5页Chinese Journal of Hematology

基　　金：天津市应用基础与前沿技术研究计划（16JCQNJC12200）;天津市血液病临床医学研究中心建设（15ZXLCSY00010）;中国医学科学院医学与健康科技创新工程（2016-12M-1-001、2016-12M3-004）

摘　　要：目的探讨NPM1基因表达对急性髓系白血病（AML）细胞系的影响及其机制。方法选取AML细胞系U937和HL-60细胞，转染NPM1质粒至细胞系构建稳定克隆，采用Westernblot法鉴定高表达NPM1蛋白的单克隆细胞。MTT法检测细胞增殖活性，流式细胞术检测细胞周期分布和细胞凋亡率，显微镜下计数检测集落形成能力，Westernblot法检测细胞周期相关信号通路蛋白表达．实时荧光定量PCR（RQ-PCR）法检测初诊AML患者骨髓单个核细胞NPM1基因表达水平。结果@U937和HL．60细胞中NPM1高表达组相对细胞增殖率与对照组相比，差异无统计学意义（4．681．28对3．89：50．81，3．34±0．37对2．6820．29，P值均〉0．05）。@U93V和HL-60细胞中NPM1高表达组S期细胞比例均明显高于对照组[（50．22±3．42）％对（39．78±3．80）％，（59．01±3．27）％对（43．94±2．08）％，P值均〈O．05]。③U937细胞NPM1高表达组和对照组相比具有更强的抗凋亡能力[（48．67±3．22）％和（68．77±10．21）％，P〈0．05]和集落形成能力（772．7：k24．0和652．3±16．5，P〈0．05），而HL．60细胞相应的两组细胞上述能力均相似。④NPM1高表达组细胞中CDK4、CyclinD1、CyclinD2及CyclinE表达明显高于对照组，而CyclinD3表达明显低于对照组。⑤细胞遗传学预后良好组AML患者NPM1定量水平低于预后中等组。结论NPM1蛋白能够促进更多的细胞进入s期，增强抗凋亡和细胞集落形成能力。NPMI定量水平可能预示细胞遗传学的危险度。Objective To investigate the impact and mechanism of NPM1 gene expression on acute myeloid leukemia （AML） cell lines. Methods Human AML cell line U937 and HL-60 cells were transfected with NPM1 plasmid to establish stable clones, and the high NPM1 protein expression （NPM1hi） clones were screened by Western blot. The cell proliferation was assayed by methylthiazolyl tetrazolium bromide （MTT）, cell cycle and cell apoptosis by flow cytometric, cell colony formation by microscope count, the molecular pathways related to cell cycle by Western blot. The expression of NPM1 gene in primary AML bone marrow mononuclear cells （BMMC） was investigated by RQ-PCR. Results （1）The proliferation of NPM1hi U937 and HL-60 cells was similar with that of control cells （4.68± 1.28 vs 3.89± 0.81, 3.34±0.37 vs 2.68±0.29, P 〉 0.05）. （2）The percentage of S phase in NPMYi U937 and HL-60 cells was higher than that in control cells [ （50.22：E3.42）% vs （39.78±3.80）%, （59.01±3.27）% vs （43.94±2.08）%, P 〈 0.05]. （3）The anti-apoptosis capacity and colony formation abilities of NPM1hi U937 cells increased than that of control cells [ （68.8±10.2）% vs （48.7±3.22）%, and （772.7±24.0） vs （652.3±16.5）, P〈 0.05], but the above abilities of NPMP HL60 cells were similar with that of control cells.（4） The expressions of CDK4, Cyclin D1, Cyclin D2 and Cyclin E in NPM1hi leukemia cells were higher than thatof control cells, but the expression of Cyclin D3 was lower. （5）The NPM1 expression levels in AML patients with favorable cytogenetic prognosis were lower than that of patients with intermediate prognosis. Conclusions NPM1 protein could promote more cells to enter S phase, enhance the ability of anti- apoptosis and colony formation in AML cell lines. The quantitative level of NPM1 may predict the cytogenetic risk of AML patients.

关 键 词：基因 NPM1 白血病 髓样 急性 

分 类 号：R733.71[医药卫生—肿瘤]