细胞分化剂Ⅱ诱导HL-60细胞的分化及其机制

Effect and mechanism of cell differentiation agent Ⅱ on the differentiation of HL-60 cells

目的探讨细胞分化剂II（CDA-Ⅱ）对人类急性早幼粒细胞白血病细胞株HL-60分化的影响及其机制。方法CDA-Ⅱ作用HL-60后，采用瑞特染色法观察细胞的形态变化；流式细胞术检测CD11b阳性细胞比例；用表达谱芯片检测差异表达基因。结果瑞特染色显示CDA-Ⅱ可诱导HL-60细胞向髓系成熟阶段分化，且呈时间依赖性。HL-60细胞经CDA-Ⅱ处理后CD11b阳性表达率呈时间、浓度依赖性增高。CDA-Ⅱ可引起HL-60细胞基因表达变化，其中表达变化相差2倍以上的基因有113条上调基因及140条下调基因。上调基因主要参与矿物质吸收、补体与凝血系统等6条通路；下调基因主要参与嘧啶代谢、RNA聚合酶、嘌呤代谢等9条通路。结论CDA-Ⅱ可诱导HL-60细胞基因表达变化及细胞分化，CDA-Ⅱ具有用于诱导分化治疗急性早幼粒细胞白血病的可行性。

Objective To investigate the effect and mechanism of cell differentiation agent Ⅱ （CDA- Ⅱ ） on the differentiation of human acute mycloid leukemia （APL） HL-60 cells. Methods The cell morphology and differentiation was detected by Wright-Giemsa staining, the expression of cell surface differentiation antigen CD11b of HL-60 was detected by flow cytometry, the differentially expressed genes were screened by gene expression profile chip （NimbleGen）. Results The result of Wright-Giemsa staining showed that CDA-Ⅱ induced HL-60 differentiation towards mature stages in a time-dependent manner. After treated with CDA-Ⅱ, the percentage of CD11b-positive HL-60 cells significantly increased in a time- and dose-dependent manner. The result of gene expression profiling indicated differentially expressed genes including 113 up-regulation genes and 140 down-regulation genes. The up-regulation expression genes involved in six pathways including mineral absorption, complement and coagulation cascades, down-regulation expression genes involved in nine pathways including pyrimidine metabolism, RNA polymerase, purine metabolism and so on. Conclusion CDA-Ⅱ can induce HL-60 differentiation and make gene differentially expressed. The data provide the feasibility of CDA- Ⅱ in differentiation induction therapy for APL.