摘要
核仁磷酸蛋白(nucleophosmin,NPM1)突变是近年发现的在急性髓系白血病中发挥重要作用的基因改变,为探讨NPM1突变对K562白血病细胞体外增殖和侵袭能力的影响,将载体pEGFPC1-NPM1-mA转染K562细胞系,构建稳定表达NPM1突变蛋白的白血病细胞株(K562-mA)。利用细胞生长曲线观察细胞体外增殖能力;流式细胞仪检测细胞周期进程改变;细胞粘附、Transwell实验分别用以观察细胞体外粘附、迁移及侵袭能力。结果发现,NPM1突变转染后K562细胞体外增殖能力明显减弱;同时G1期细胞比例明显增高,S期细胞比例显著减低。与未处理组和空载体转染组细胞相比,K562-mA细胞体外迁移能力有所增加,但细胞粘附及侵袭能力却明显减弱。提示NPM1突变基因的表达能够抑制白血病细胞体外增殖和侵袭能力,为进一步深入探讨NPM1突变在白血病发生发展中的调控机制奠定了良好的基础。
Nucleophosmin（NPM1）mutations,described recently,play an important role in acute myeloid leukemia.To investigate the role of NPM1 mutations in K562 leukemic cell proliferation and invasion in vitro,the pEGFPC1-NPM1-mA plasmid vector with NPM1 mutation A（NPM1-mA）was transfected into K562 cells,and the K562-mA cells with stably expressed NPM1-mA protein were established.Cell growth curve was used to determine the proliferation potential in vitro.FCM was used to detect the cell cycle.Cell adhesion assay and Transwell were used to detect the adhesion,migration and invasion capability.The results showed that the proliferation potential of K562-mA cells was decreased.Compared with control groups,the percentage of cells in the G1 phase was increased remarkably and that in the S phase was decreased significantly.In addition,the migration ability of K562-mA cells was enhanced,but the adhesion and invasion capability of cells were attenuated.NPM1 mutations may inhibit K562 cells proliferation and invasion in vitro,which provide a better foundation for studying the molecule mechanisms of NPM1 mutations in leukemiagenesis.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第11期6-10,共5页
China Biotechnology
基金
国家自然科学基金(30872418)
重庆市科委自然科学基金(CSTC
2010BB5363)资助项目
