Andreeff M, Tafuri A, Hegewisch-Becker S
Leukemia Cell Biology Laboratory, Memorial Sloan-Kettering Cancer Center, Cornell University Medical College New York, NY 10021.
Haematol Blood Transfus. 1990;33:747-62. doi: 10.1007/978-3-642-74643-7_137.
Prognostic models for acute myeloid and lymphoid leukemias are presented that demonstrate that cell kinetic quiescence in acute leukemia is associated with poor response to chemotherapy, short remission duration, and survival. Recruitment of cells into the cell cycle should therefore enhance cytotoxic effects of cell cycle - specific chemotherapeutic agents. We previously demonstrated recruitment of myeloid leukemic cells by cytokines. We have now investigated whether recruitment can be used to increase cell killing by cytosine arabinoside (Ara-C). Blast cells from 16 acute leukemias were stimulated with cytokines as follows: 13 acute myeloid leukemias (AML) and 3 chronic myeloid leukemia (CML) in blastic phase (1 lymphoid, 2 myeloid) were treated with recombinant human granulocyte colony stimulating factor (rhG-CSF), recombinant human granulocyte-macrophage colony stimulating factor (rhG-CSF, AMGEN, 500 U/ml each), and recombinant human interleukin-3 (rhIL-3, IMMUNEX, 20 ng/ml), alone and in combination. After 48 h, at the time of maximal DNA synthesis, Ara-C (10(-3) M) was added and cell counts, cytokinetics (DNA/RNA, DNA/bromodeoxyuridine and DNA/Ki67 flow cytometry), and cell viability/clonogenicity (fluorescein diacetate/propidium iodide exclusion flow cytometry) were investigated. In all 13 cases of AML recruitment was found; in 6 of these cases over a three fold increase in S phase (P = 0.008) and a significant (P = 0.004) depletion of G0 was demonstrated. In 9 of 13 patients with AML, the effect of Ara-C was investigated, and in 3 of 5 patients with over three fold increase in S phase, Ara-C toxicity was enhanced. None of the patients with less than a three fold increase in S phase and no demonstrable recruitment from G0 had increased Ara-C cytotoxicity. Ara-C cytoreduction was paralled by reduction in clonogenicity as demonstrated by fluorescein diacetate/propidium iodide (FDA/PI) flow cytometry. Four samples of acute lymphoblastic leukemia (ALL) were treated with low molecular weight B-cell growth factor (15 kDa) and recruitment of aneuploid cells from G0 to G1 was found in all patients (from 19.3% to 84.9%). These results indicate that recruitment of leukemic cells is inducible by cytokines and that the cytotoxicity of cell cycle-specific drugs such as Ara-C can be increased. This concept is presently being tested in vivo.
本文提出了急性髓系白血病和急性淋巴细胞白血病的预后模型，该模型表明急性白血病中的细胞动力学静止与化疗反应不佳、缓解期短和生存率低有关。因此，将细胞募集到细胞周期中应能增强细胞周期特异性化疗药物的细胞毒性作用。我们之前证明了细胞因子可募集髓系白血病细胞。我们现在研究了募集是否可用于增强阿糖胞苷（Ara-C）的细胞杀伤作用。用细胞因子刺激16例急性白血病的原始细胞，具体如下：13例急性髓系白血病（AML）和处于急变期的3例慢性髓系白血病（CML）（1例淋巴细胞性、2例髓细胞性）分别单独或联合使用重组人粒细胞集落刺激因子（rhG-CSF）、重组人粒细胞-巨噬细胞集落刺激因子（rhGM-CSF，安进公司，各500 U/ml）和重组人白细胞介素-3（rhIL-3，免疫ex公司，20 ng/ml）进行治疗。48小时后，在DNA合成达到最大值时加入Ara-C（10⁻³ M），并研究细胞计数、细胞动力学（DNA/RNA、DNA/溴脱氧尿苷和DNA/Ki67流式细胞术）以及细胞活力/克隆形成能力（荧光素二乙酸酯/碘化丙啶排除流式细胞术）。在所有13例AML病例中均发现了细胞募集；其中6例S期增加了三倍以上（P = 0.008），且G0期显著减少（P = 0.004）。在13例AML患者中的9例中研究了Ara-C的作用，在S期增加了三倍以上的5例患者中的3例中，Ara-C毒性增强。S期增加不到三倍且未发现从G0期有明显募集的患者中，没有一例Ara-C细胞毒性增加。通过荧光素二乙酸酯/碘化丙啶（FDA/PI）流式细胞术证明，Ara-C的细胞减少与克隆形成能力的降低平行。4例急性淋巴细胞白血病（ALL）样本用低分子量B细胞生长因子（15 kDa）进行治疗，所有患者均发现非整倍体细胞从G0期募集到G1期（从19.3%至84.9%）。这些结果表明，细胞因子可诱导白血病细胞的募集，并且细胞周期特异性药物如Ara-C的细胞毒性可以增加。这一概念目前正在体内进行测试。
