Regulatory properties and cellular redistribution of zinc during macrophage differentiation of human leukemia cells
Regulatory properties and cellular redistribution of zinc during macrophage differentiation of human leukemia cells
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DOI：
10.1016/j.jsb.2005.09.012
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发表时间：
2006-07-01
影响因子：
3
通讯作者：
Huberman, Eliezer
中科院分区：
文献类型：
--
作者：
Glesne, David;Vogt, Stefan;Huberman, Eliezer
许多蛋白质需要微量金属的结合，如钙、铁、铜或锌，这可能会调节它们的结构、功能或活性。为了确定巨噬细胞分化过程中是否存在金属蛋白分布或金属浓度的整体变化，我们用phorbol 12-肉豆酸酯13-乙酸酯（PMA）诱导人髓系HL-60白血病细胞，并使用硬x射线荧光显微分析定量绘制其金属含量。我们发现在HL-60分化诱导的早期，细胞核中锌含量有短暂的增加。这一发现在HL-60细胞和U-937白血病细胞中得到了证实。通过检测pkc - β缺失的HL-60变体HL-525和通过稳定过表达恢复pkc - β的HL-525细胞的锌含量，确定了蛋白激酶c - β (pkc - β)在这一过程中的作用。Cu和Zn的化学螯合可以抑制HL-60细胞的巨噬细胞分化，表明在这一过程中需要这些金属。最后，我们证明了HL-60细胞在低锌环境下的生长消除了它们对pma诱导的分化的敏感性，并且这种能力可以通过添加外源锌来部分恢复。(c) 2006年Elsevier Inc.出版
Many proteins require the binding of trace metals such as Ca, Fe, Cu, or Zn, which may modulate their structure, function, or activity. To determine if there were any overall changes in metalloprotein distribution or metal concentration during the process of macrophage differentiation we induced human myeloid HL-60 leukemia cells with phorbol 12-myristate 13-acetate (PMA) and quantitatively mapped their metal content using hard X-ray fluorescence micro-analysis. We found a transient increase in the zinc content of HL-60 cell nuclei during the early stages of differentiation induction. This finding was confirmed by spectrofluorometry in HL-60 cells and extended to U-937 leukemia cells. A role for protein kinase C-beta (PKC-beta) in this process was established by examining zinc content in an HL-60 variant, HL-525, which is PKC-beta deficient, and in HL-525 cells in which PKC-beta was restored by stable overexpression. Chemical chelation of both Cu and Zn served to inhibit macrophage differentiation in HL-60 cells, indicating a requirement for these metals during this process. Finally, we demonstrate that growth of HL-60 cells in a low-zinc environment removes their susceptibility to PMA-induced differentiation, and that this capacity can be partially restored by the addition of exogenous zinc. (c) 2006 Published by Elsevier Inc.