Kataoka T, Morishita Y, Ogura M, Morishima Y, Towatari M, Kato Y, Inoue H, Saito H
First Department of Internal Medicine, Nagoya University School of Medicine, Japan.
Cancer Res. 1990 Dec 1;50(23):7703-9.
A novel human myeloid leukemia cell line, NKM-1, was established from a patient with acute myeloid leukemia (FAB classification M2). The cells were positive for myeloperoxidase staining and cluster of differentiation 15 cell surface antigen. Radiolabeled recombinant human granulocyte (G) colony-stimulating factor (CSF) was used, and 60 specific binding sites/cell with a Kd 100 pmol/liter were demonstrated on the cell surface. 125I-G-CSF binding was not inhibited by interleukin-3, granulocyte-macrophage CSF, or macrophage (M) CSF. NKM-1 cells also expressed M-CSF receptors detected by c-fms mRNA expression. In concordance with the receptor expression, NKM-1 cells proliferated in response to exogenous G-CSF or M-CSF in a dose-dependent manner (0.1-100 ng/ml), while interleukin-3 or granulocyte-macrophage CSF had no effect. Colony-forming capacity of NKM-1 cells in semisolid agar was also enhanced with the addition of 10 ng/ml of G-CSF or M-CSF but decreased at higher concentrations. During CSF stimulation, no remarkable changes were observed morphologically and phenotypically. The stimulatory effect of G-CSF and M-CSF on the cell growth was additive. Neither G-CSF-binding capacity nor c-fms mRNA expression was altered by pretreatment with M-CSF or G-CSF, respectively. This cell line may provide a useful in vitro model for the study of CSF roles in myeloid leukemia cell proliferation.
一种新的人类髓系白血病细胞系NKM-1,是从一名急性髓系白血病(FAB分类M2)患者中建立的。这些细胞髓过氧化物酶染色和分化簇15细胞表面抗原呈阳性。使用放射性标记的重组人粒细胞(G)集落刺激因子(CSF),在细胞表面检测到60个特异性结合位点/细胞,解离常数(Kd)为100 pmol/升。125I-G-CSF结合不受白细胞介素-3、粒细胞-巨噬细胞集落刺激因子或巨噬细胞(M)集落刺激因子的抑制。NKM-1细胞还通过c-fms mRNA表达检测到表达M-集落刺激因子受体。与受体表达一致,NKM-1细胞对外源G-集落刺激因子或M-集落刺激因子以剂量依赖方式(0.1-100 ng/ml)增殖,而白细胞介素-3或粒细胞-巨噬细胞集落刺激因子则无作用。在半固体琼脂中添加10 ng/ml的G-集落刺激因子或M-集落刺激因子可增强NKM-1细胞的集落形成能力,但在较高浓度下则降低。在集落刺激因子刺激期间,未观察到形态和表型上的明显变化。G-集落刺激因子和M-集落刺激因子对细胞生长的刺激作用是相加的。分别用M-集落刺激因子或G-集落刺激因子预处理,G-集落刺激因子结合能力和c-fms mRNA表达均未改变。该细胞系可能为研究集落刺激因子在髓系白血病细胞增殖中的作用提供一个有用的体外模型。