Cook N, Dexter T M, Lord B I, Cragoe E J, Whetton A D
Department of Biochemistry and Applied Molecular Biology, UMIST, UK.
EMBO J. 1989 Oct;8(10):2967-74. doi: 10.1002/j.1460-2075.1989.tb08446.x.
We have prepared a population of bone marrow cells that is highly enriched in neutrophil/macrophage progenitor cells (GM-CFC). Four distinct haemopoietic growth factors can stimulate the formation of mature cells from this population, although the proportions of neutrophils and/or macrophages produced varied depending on the growth factor employed: interleukin 3 (IL-3) and granulocyte/macrophage colony-stimulating factor (GM-CSF) stimulated the formation of colonies containing both neutrophils and macrophages; macrophage colony-stimulating factor (M-CSF) produced predominantly macrophage colonies; and granulocyte colony-stimulating factor (G-CSF) promoted neutrophil colony formation. Combinations of these four growth factors did not lead to any additive or synergistic effect on the number of colonies produced in clonal soft agar assays, indicating the presence of a common set of cells responsive to all four haemopoietic growth factors. These enriched progenitor cells therefore represent an ideal population to study myeloid growth-factor-stimulated survival, proliferation and development. Using this population we have examined the molecular signalling mechanisms associated with progenitor cell proliferation. We have shown that modulation of cyclic AMP levels has no apparent role in GM-CFC proliferation, whereas phorbol esters and/or Ca2+ ionophore can stimulate DNA synthesis, indicating a possible role for protein kinase C activation and increased cytosolic Ca2+ levels in the proliferation of these cells. The lack of ability of all four myeloid growth factors to mobilize intracellular Ca2+ infers that these effects are not achieved via inositol lipid hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)
我们制备了一群高度富集中性粒细胞/巨噬细胞祖细胞(GM-CFC)的骨髓细胞。四种不同的造血生长因子可刺激这群细胞形成成熟细胞,不过根据所使用的生长因子不同,产生的中性粒细胞和/或巨噬细胞比例有所变化:白细胞介素3(IL-3)和粒细胞/巨噬细胞集落刺激因子(GM-CSF)刺激形成同时含有中性粒细胞和巨噬细胞的集落;巨噬细胞集落刺激因子(M-CSF)主要产生巨噬细胞集落;而粒细胞集落刺激因子(G-CSF)促进中性粒细胞集落形成。在克隆软琼脂试验中,这四种生长因子的组合对产生的集落数量没有产生任何相加或协同效应,表明存在一组对所有四种造血生长因子都有反应的共同细胞。因此,这些富集的祖细胞是研究髓系生长因子刺激的存活、增殖和发育的理想细胞群。利用这群细胞,我们研究了与祖细胞增殖相关的分子信号机制。我们发现,环磷酸腺苷水平的调节在GM-CFC增殖中没有明显作用,而佛波酯和/或钙离子载体可刺激DNA合成,表明蛋白激酶C激活和胞质钙离子水平升高在这些细胞的增殖中可能发挥作用。所有四种髓系生长因子都无法动员细胞内钙离子,这表明这些效应不是通过肌醇脂质水解实现的。(摘要截短于250词)