Fischkoff S A, Hoessly M C, Rossi R M
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104-4283.
Leukemia. 1990 Apr;4(4):302-6.
HL-60 human leukemia cells undergo terminal differentiation when cultured with butyric acid. To produce cells resistant to the maturation-inducing effects of butyric acid, two strategies were followed. First, HL-60 cells were mutagenized and cultured in soft agar with inducing concentrations of butyric acid. Four clones were isolated resistant to a wide variety of differentiation inducers, including butyric acid. Second, HL-60 cells were cultured in gradually increasing concentrations of butyric acid until a normal growth rate was achieved in medium containing greater than 1 mM butyric acid. These cells retained their ability to be induced to mature to neutrophils with dimethylsulfoxide, retinoic acid, and actinomycin D; and to monocyte/macrophage like cells with tetradecanoylphorbol acetate and 1,25-(OH)2 vitamin D3. However, they no longer underwent terminal differentiation when butyric acid was added in increasing concentration, even when cytotoxic concentrations were used. The mutagenized clones cells appeared permanently butyrate resistant, but the selected clones reverted to the wild-type state when grown in the absence of butyric acid. The selected cells continued to express myeloperoxidase; the mutagenized lines did not. Thus, by using two different protocols, HL-60 cells resistant to the cytotoxic and maturation-inducing effects of butyric acid can be produced that have varied phenotypic characteristics.
HL-60人白血病细胞在与丁酸一起培养时会发生终末分化。为了产生对丁酸成熟诱导作用具有抗性的细胞，采用了两种策略。首先，对HL-60细胞进行诱变，并在含有诱导浓度丁酸的软琼脂中培养。分离出四个对多种分化诱导剂（包括丁酸）具有抗性的克隆。其次，将HL-60细胞在逐渐增加浓度的丁酸中培养，直到在含有大于1 mM丁酸的培养基中达到正常生长速率。这些细胞保留了用二甲基亚砜、视黄酸和放线菌素D诱导成熟为中性粒细胞的能力；以及用十四酰佛波醇乙酸酯和1,25-(OH)2维生素D3诱导成熟为单核细胞/巨噬细胞样细胞的能力。然而，当加入浓度不断增加的丁酸时，即使使用细胞毒性浓度，它们也不再经历终末分化。诱变克隆细胞似乎对丁酸具有永久抗性，但所选克隆在无丁酸的条件下生长时会恢复到野生型状态。所选细胞继续表达髓过氧化物酶；诱变株系则不表达。因此，通过使用两种不同的方案，可以产生对丁酸的细胞毒性和成熟诱导作用具有抗性且具有不同表型特征的HL-60细胞。
