Induction of HL-60 leukemia cell differentiation by the novel antifolate 5,10-dideazatetrahydrofolic acid.
Induction of HL-60 leukemia cell differentiation by the novel antifolate 5,10-dideazatetrahydrofolic acid.
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发表时间：
1989-09
期刊：
影响因子：
11.2
通讯作者：
J. Sokoloski;G. Beardsley;A. Sartorelli
中科院分区：
文献类型：
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作者：
J. Sokoloski;G. Beardsley;A. Sartorelli
新的四氢叶酸，5，10-二脱氮四氢叶酸（DDATHF），被设计为在二氢叶酸还原酶以外的位点抑制叶酸代谢。DDATHF已被证明能抑制甘氨酰胺核糖核苷酸转化酶，这是一种需要叶酸的酶，催化从头嘌呤核苷酸生物合成途径中两个一碳转移反应中的第一个。HL-60早幼粒细胞白血病细胞与5 × 10（-8）至10（-5）M DDATHF孵育48小时后，生长受到明显抑制，第4天细胞复制完全停止。DDATHF处理的HL-60细胞的细胞周期分析表明，在早期S期的第3天，随后在细胞周期的G1和G2 + M期的细胞积累的初始块。生长抑制伴随着浓度依赖性增加的成熟髓系细胞的百分比，表达硝基四唑蓝阳性，和非特异性酯酶活性的小幅增加。次黄嘌呤和5（4）-氨基-4（5）-咪唑甲酰胺完全阻止DDATHF诱导分化和抑制生长，表明细胞内嘌呤核苷酸库的消耗在这种抑制剂的生物学效应中具有重要作用。DDATHF在2小时内引起细胞内GTP和ATP水平的显著降低，在24小时观察到最大降低，这是该试剂抑制细胞增殖之前的时间间隔，这一发现证实了这种可能性。嘧啶核苷三磷酸水平显着增加，在这些条件下。这些结果表明，嘌呤核苷酸对DDATHF抑制HL-60白血病细胞生长和诱导分化的重要性。
The novel tetrahydrofolate, 5,10-dideazatetrahydrofolic acid (DDATHF), was designed as an inhibitor of folate metabolism at a site other than dihydrofolate reductase. DDATHF has been shown to inhibit glycinamide ribonucleotide transformylase, a folate-requiring enzyme that catalyzes the first of two one-carbon transfer reactions in the de novo purine nucleotide biosynthetic pathway. Incubation of HL-60 promyelocytic leukemia cells with 5 x 10(-8) to 10(-5) M DDATHF resulted in a marked inhibition of growth after 48 h, with a complete cessation of cellular replication by day 4. Cell cycle analyses of DDATHF-treated HL-60 cells demonstrated an initial block in early S phase by day 3 followed by an accumulation of cells in the G1 and G2 + M phases of the cell cycle. Inhibition of growth was accompanied by a concentration-dependent increase in the percentage of mature myeloid cells that expressed nitroblue tetrazolium positivity, and a small increase in nonspecific esterase activity. Induction of differentiation and inhibition of growth by DDATHF were completely prevented by hypoxanthine and 5(4)-amino-4(5)-imidazole carboxamide, suggesting that depletion of intracellular purine nucleotide pools has an important role in the biological effects of this inhibitor. This possibility was confirmed by the finding that DDATHF caused a pronounced reduction in intracellular GTP and ATP levels within 2 h, with maximum decreases being observed by 24 h, a time interval which preceded the inhibition of cellular proliferation by this agent. Pyrimidine nucleoside triphosphate levels were markedly increased under these conditions. The findings indicate the importance of purine nucleotides to both the inhibition of growth and the induction of differentiation of HL-60 leukemia cells by DDATHF.