NB4细胞株分化中NDRG1表达的研究

Study on the expression of NDRG1 in induced differentiation of NB4 cell line

目的:应用多种分化诱导剂处理急性早幼粒细胞白血病(APL)细胞系NB4细胞株,探讨NDRG1表达在白血病细胞诱导分化中的作用。方法:采用1×10^(-7)mol/L全反式维甲酸(ATRA)、1%二甲基亚砜(DMSO)和20 nmol/L佛波酯(TPA)分别处理NB4细胞48h,并通过细胞染色、流式细胞检测、蛋白印迹(Western blot)和实时PCR分别检测分化细胞的核型、CD11b、CD11c、CD14表达,观察ndrgl mRNA与蛋白的变化。结果:3种分化诱导剂分别处理细胞株后,细胞核缩小且呈分叶状改变,细胞内NDRGl蛋白表达均呈时间依赖性上调;经ATRA和DMSO处理的细胞,其ndrgl mRNA发生上调,而经TPA处理的细胞却未发生ndrgl mRNA表达上调。结论:细胞信号调控分子NDRG1与细胞分化密切相关。

Objective To treat acute promyelocytic leukemia （APL） cell line NB4 by several differentiation inducers and to investigate the role played by N-myc downstream-regulated gene 1 （NDRG1） expression in the differentiation induction of APL cells. Methods All-trans retinoic acid （ATRA） 10^-7 mol/L, 1% dimethyl sulfoxide （DMSO） and 12-O-tetradecanoylphorbol-13-acetate （TPA） 20 nmol/L were used to treat NB4 cells for 48 hours, respectively. Cell morphological features were examined after Wright＇s staining of cells; cell surface antigen CD11b, CD11c and CD14 were analyzed by flow cytometry; NDRG1 protein expression was measured by Western blot and NDRG1 mRNA was assayed by real-time PCR. Results After treated with these three differentiation inducers, NB4 cells showed a decreased karyoplasmic ratio. The expression of NDRG1 protein was upregulated with a dependence on time. The expression of NDRG1 mRNA was upregulated when treated with ATRA and DMSO, but not when treated with TPA. Conclusions Cell signaling modulating factor NDRG1 has close correlation with cell differentiation.