基础医学与临床 ›› 2012, Vol. 32 ›› Issue (3): 251-256.

• 研究论文 • 上一篇    下一篇

NPM1基因突变表达对THP-1白血病细胞增殖和凋亡的作用探讨

覃凤娴1,邵会媛陈先春1,谭诗张慧娟张伶1   

  • 收稿日期:2011-04-08 修回日期:2011-10-08 出版日期:2012-03-05 发布日期:2012-02-27
  • 通讯作者: 覃凤娴 E-mail:ypp19850325@126.com
  • 基金资助:
    国家重点基础研究发展计划;重庆市科委自然科学基金

The role of nucleophosmin (NPM1) gene mutations in the proliferation and apoptosis of THP-1 leukemic cells

  • Received:2011-04-08 Revised:2011-10-08 Online:2012-03-05 Published:2012-02-27
  • Contact: Feng-Xian QIN E-mail:ypp19850325@126.com

摘要: 摘要:目的 探讨核仁磷酸蛋白(NPM1)基因突变对白血病细胞增殖潜能和凋亡发生的影响。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 A型突变蛋白的白血病细胞株(THP-1 mA),同时设立未处理组(THP-1)和空载体转染组(THP-1 C1)作为对照。通过MTT试验观察细胞体外增殖能力,流式细胞术分析细胞周期分布及凋亡发生率的改变;采用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 与未处理组和空载体转染组细胞相比较,携NPM1突变体的THP-1 mA细胞体外增殖能力明显增强,S期细胞比例明显增高,G1期细胞比例显著减低;而NPM1突变体转染后THP-1细胞的凋亡率没有显著变化,同时细胞凋亡相关蛋白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,这为进一步阐明NPM1突变与AML的关系提供了新的科学依据。

关键词: 关键词:NPM1突变, 白血病, THP-1细胞系, 增殖, 凋亡

Abstract: Abstract: 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. Addition, the percentage of cells in the S phase was increased significantly and that in the G1 remarkably decreased. While there was no significantly change on cellular apoptosis, and no significantly difference was observed on the expression of Bax and Bcl-2, no matter on the mRNA level or protein level. The ratio of Bax/Bcl-2 also had no significantly change. Conclusion NPM1 mutations may promote cells proliferation potential in vitro, but had no significantly influence on cellular apoptosis, which may provide a new scientific evidence for the relationship of NPM1 mutation and AML.

Key words: Key words: NPM1 mutation, leukemia, THP-1 cell line, proliferation, apoptosis

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