目的:探讨WT1、RbAp46及IGFBP-rP1基因与NB4细胞诱导分化的关系。方法:利用实时定量RT—PCR方法检测NB4细胞诱导分化不同时间点WT1、RbAp46和IGFBP-rP1基因的表达水平,并用流式细胞仪检测NB4细胞表面抗原CD11b的变化。结果:随着NB4细胞向粒系终末分化,WT1、jGFBP-rP1基因的表达水平迅速下降。0.5μmol/L全反式维甲酸(all transretinoic acid,ATRA)作用0、4、8、12、24与48h后WT1N分别为1.91、1.21、0.60、0.44、0.18与0.04;IGFBP-rP1N分别为1.10、0.80、0.54、0.28、0.17与0.15。RbAp46基因的平均表达水平则下降缓慢,分别为2.65、1.86、1.40、1.27、1.48与0.49。NB4细胞处理组WT1基因的改变分别与RbAp46和IGFBP-rP1基因的变化存在相关性(r=0.829,P=0.021;r=1,P〈0.001),三者均与CD11b的变化呈负相关(r=-1.0,P〈0.001;r=-0.829,P=0.021与r=-1.0,P〈0.001)。结论:WT1、RbAp46和IGFBP-rP1基因的高表达可能参与了阻滞NB4细胞分化的过程。
Objective:To investigate the relationship between the expression level of WT1, RbAp46, and IGFBP-rP1 genes and the differentiation of NB4 leukemic cells. Methods: Real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method was used for detecting the expressions of WT1, RbAp46, and IGFBP-rP 1 genes in NB4 leukemic cells at different time points after treatment with 0.5 μmol/L ATRA(all transretinoic acid). The expression of CDllb was simultaneously detected using flow cytometry. Results:WT1 and IGFBP-rP 1 gene expressions rapidly decreased during the differentiation of NB4 cells to the end stage. The average expression values of WT1N were 1.91, 1.21, 0.60, 0.44, 0.18, and 0.04 at 0, 4, 8, 12, 24, and 48 h after exposure to ATRA, respectively. The average expression values of IGFBP-rPIN were 1.10, 0.80, 0.54, 0.28, 0.17, and 0. 15, respectively, at same time points. The expression of RbAp46N decreased slowly. The average values were 2.65, 1.86, 1.40, 1.27, 1.48, and 0.49, respectively. The change of WT1 gene expression in treatment group correlated to the change of RbAp46 and IGFBP-rP1 gene expressions (r= 0. 829, P=0. 021 ; r= 1, P〈0. 001). The changes of the expressions of three genes were negatively related to that of CDllb expression (r=-1.0, P〈0.001; r=-0.829, P= 0. 021 ; r= -1.0, P〈0. 001). Conclusion:The high expressions of WT1, RbAp46, and IGFBP-rP1 genes were probably involved in the blockage of differentiation of NB4 cells.