Action of granulocyte-macrophage colony-stimulating factors: studies using a human leukemia cell line.
Action of granulocyte-macrophage colony-stimulating factors: studies using a human leukemia cell line.
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粒细胞-巨噬细胞集落刺激因子的作用：使用人类白血病细胞系的研究。
DOI：
10.1073/pnas.77.9.5346
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发表时间：
1980
影响因子：
11.1
通讯作者：
Koeffler,HP
中科院分区：
文献类型：
--
作者：
Lusis,AJ;Koeffler,HP
粒细胞-巨噬细胞集落刺激因子（CSF）先前已被证明可以刺激软琼脂培养物中KG-1人髓性白血病细胞系的集落形成。我们已经使用KG-1细胞作为模型系统来研究CSF与髓系细胞的相互作用。我们现在报道KG-1细胞在液体培养中暴露于人CSF导致RNA合成的快速（3小时内）爆发，并且在约10小时的滞后之后，刺激DNA和蛋白质合成。RNA和蛋白质合成最大刺激约2倍，DNA合成刺激约2.5倍。刺激是特异性的;各种生长因子、激素和小鼠CSF对KG-1大分子合成没有影响。用CSF治疗没有明显改变KG-1细胞（主要是成髓细胞）的形态学外观，也没有定性地影响新合成的蛋白质可通过一维和二维电泳分离的模式。在琼脂培养中对CSF无反应的几种髓性白血病细胞系，包括KG-1的去分化变体，在暴露于CSF时显示出很少或没有大分子合成的刺激。我们利用CSF依赖性刺激KG-1的大分子合成，开发了一种快速、灵敏的人CSF微量测定法。该测定法，涉及胸苷掺入的细胞，应该是有用的人CSF的表征和纯化。
Granulocyte-macrophage colony-stimulating factors (CSFs) have previously been shown to stimulate colony formation in soft agar culture by a human myelogenous leukemia cell line known as KG-1. We have used KG-1 cells as a model system to investigate the interaction of CSF with myeloid cells. We now report that exposure of KG-1 cells to human CSFs in liquid culture results in a rapid (within 3 hr) burst of RNA synthesis and, after a lag of about 10 hr, a stimulation of DNA and protein synthesis. RNA and protein synthesis were maximally stimulated about 2-fold and DNA synthesis was stimulated about 2.5-fold. The stimulation was specific; various growth factors, hormones, and mouse CSFs had no effect on KG-1 macromolecular synthesis. Treatment with CSF did not discernibly alter the morphological appearance of the KG-1 cells (primarily myeloblasts) nor did it qualitatively affect the pattern of newly synthesized proteins separable by one- and two-dimensional electrophoresis. Several myeloid leukemia cell lines that were not responsive to CSF in agar culture, including a dedifferentiated variant of KG-1, showed little or no stimulation of macromolecular synthesis upon exposure to CSF. We have used the CSF-dependent stimulation of macromolecular synthesis of KG-1 to develop a rapid, sensitive microassay for human CSFs. The assay, involving thymidine incorporation by the cells, should be useful for characterization and purification of human CSFs.