Ahmed N, Williams J F, Weidemann M J
Department of Biochemistry, Faculty of Science, Australian National University, Canberra ACT.
Biochem Int. 1991 Feb;23(3):591-602.
The release of the reactive oxygen species that accompanies the oxidative burst was studied in HL60 cells differentiated with either dimethylsulphoxide, butyrate or phorbol myristate acetate in order to establish the extent to which differentiated cells are phenotypically similar to human neutrophils, monocytes and macrophages. When phorbol myristate acetate was used as a stimulus, the rates of superoxide production by dimethylsulphoxide and butyrate differentiated HL60 cells was not significantly different from those observed in neutrophils and monocytes isolated from normal peripheral blood. Similar results were obtained when luminol-dependent chemiluminescence was measured in the presence of horseradish peroxidase using phorbol myristate acetate as the stimulus. However, in the absence of horseradish peroxidase, the luminol-dependent chemiluminescence in the dimethylsulphoxide and butyrate-differentiated HL60 cells was significantly lower than that of the control cells isolated from human blood, reflecting the absence of myeloperoxidase in the differentiated cells. In contrast, HL60 cells differentiated by phorbol myristate acetate failed to show any increased generation of superoxide or luminol-dependent chemiluminescence upon stimulation. Impaired release of lysosomal enzymes by the chemically differentiated cells suggests impairments in the extent of differentiation resulting in cells with defective azurophilic degranulation processes. It is concluded that HL60 cells differentiated by the above agents are somewhat controversial models of promyelocyte differentiation into typical neutrophilic, monocytic and macrophage-like cells.
为了确定分化细胞在表型上与人类中性粒细胞、单核细胞和巨噬细胞的相似程度，研究了用二甲基亚砜、丁酸盐或佛波酯肉豆蔻酸酯诱导分化的HL60细胞中伴随氧化爆发的活性氧释放情况。当使用佛波酯肉豆蔻酸酯作为刺激物时，二甲基亚砜和丁酸盐分化的HL60细胞产生超氧化物的速率与从正常外周血中分离出的中性粒细胞和单核细胞中观察到的速率没有显著差异。当以佛波酯肉豆蔻酸酯作为刺激物，在辣根过氧化物酶存在的情况下测量鲁米诺依赖性化学发光时，也得到了类似的结果。然而，在没有辣根过氧化物酶的情况下，二甲基亚砜和丁酸盐分化的HL60细胞中鲁米诺依赖性化学发光显著低于从人血中分离出的对照细胞，这反映了分化细胞中缺乏髓过氧化物酶。相比之下，经佛波酯肉豆蔻酸酯分化的HL60细胞在受到刺激后未能显示出超氧化物生成或鲁米诺依赖性化学发光的任何增加。化学分化的细胞中溶酶体酶释放受损表明分化程度存在缺陷，导致细胞的嗜天青颗粒脱粒过程有缺陷。得出的结论是，用上述试剂分化的HL60细胞是原粒细胞分化为典型嗜中性、单核细胞和巨噬细胞样细胞的有点有争议的模型。
