Comparison of protein, RNA, and DNA binding and cell-cycle-specific growth inhibitory effects of nickel compounds in cultured cells.
Comparison of protein, RNA, and DNA binding and cell-cycle-specific growth inhibitory effects of nickel compounds in cultured cells.
复制标题
培养细胞中镍化合物的蛋白质、RNA 和 DNA 结合以及细胞周期特异性生长抑制作用的比较。
DOI：
10.1016/0041-008x(82)90318-0
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发表时间：
1982
影响因子：
3.8
通讯作者：
M. Costa
中科院分区：
文献类型：
--
作者：
P. B. Harnett;S. Robison;D. Swartzendruber;M. Costa
晶态NiS颗粒是一种有效的形态转化诱导剂，能被细胞主动吞噬。水溶性镍化合物的效力较弱，可能是因为进入细胞的镍总量较少，其亚细胞分布在许多方面与NiS以内化颗粒形式进入后的分布不同。为了进一步研究这个问题，我们检测了从培养的中国仓鼠卵巢细胞中分离出的DNA、RNA和蛋白质与63NiS或63NiCl2处理的结合。用10μg/ml63NiS处理培养细胞3天后，镍与脱氧核糖核酸或蛋白质的结合范围为每毫克大分子结合1μ克镍。然而，用浓度为10μg/ml的63NiCl2处理细胞1-5天后，与脱氧核糖核酸、核糖核酸和蛋白质结合的镍大约减少了一到几个数量级。在63NiCl2处理的情况下，细胞蛋白中结合的63Ni大约是相应的RNA或DNA组分的100倍。然而，与同样用63NiCl2处理的细胞相比，用结晶NiS处理的细胞所获得的蛋白质组分含有更多的镍结合约15倍。与用类似的NiCl2处理的细胞相比，晶态NiS处理后RNA或DNA的结合镍多出300到2000倍。与63NiCl2与蛋白质的选择性结合相反，用晶体63NiS处理的培养细胞具有与RNA、DNA和蛋白质相关的同等水平的镍。由于63NiS晶化处理后63Ni与这些大分子的相互作用不是由于实际粒子的结合，63NiS粒子在细胞内的溶解可能对可结合的63Ni离子的分布起着重要的控制作用。与同样暴露在63NiCl2中的培养物相比，用63NiS处理细胞产生了更强的细胞内大分子结合，晶体NiS在减缓和阻止细胞增殖方面的作用更强。晶态NiS在比NiCl2类似地阻止细胞生长所需的浓度低得多的浓度下引起明显的细胞周期特异性阻断。流式细胞仪分析显示，两种化合物均选择性地将细胞周期阻滞在S期(DNA合成期)。这些结果提示了这两种化合物固有的共同毒性机制和位置，并与DNA复制的细胞生长阶段有关。
Crystalline NiS particles are potent inducers of morphological transformation and are actively phagocytosed by cells. Water-soluble nickel compounds are less potent, possibly because the total amount of nickel that enters cells is less, and its subcellular distribution differs in a number of ways from that following the entry of NiS in the form of an internalized particle. To further study this problem, we have examined the binding of63Ni to DNA, RNA, and protein isolated from cultured Chinese hamster ovary cells treated with either crystalline63NiS or63NiCl2. Treatment of cultured cells with63NiS at 10 μg/ml for 3 days resulted in binding of nickel to DNA, RNA, or protein in the range of 1 μg of nickel bound per milligram macromolecule. However, similar treatment of cells with63NiCl2at 10 μg/ml for 1 to 5 days resulted in approximately one to several orders of magnitude less nickel bound to DNA, RNA, and protein. In the case of63NiCl2treatment, cellular proteins contained about 100 times more63Ni bound than the respective RNA or DNA fractions. However, the protein fraction obtained from cells treated with crystalline NiS contained about 15 times more nickel bound than the same fraction isolated from cells that were similarly treated with63NiCl2. RNA or DNA had 300 to 2000 times more bound nickel following crystalline NiS treatment compared to cells treated similarly with NiCl2. In contrast to the selective binding of63NiCl2to protein, cultured cells treated with crystalline63NiS had equivalent levels of nickel associated with RNA, DNA, and protein. Since the interaction of63Ni with these macromolecules following crystalline63NiS treatment was not due to the binding of the actual particles, the dissolution of intracellular63NiS particles probably plays an important role in governing the distribution of63Ni ions available for binding. The greater intracellular macromolecular binding of63Ni resulting from treatment of cells with63NiS compared to cultures similarly exposed to63NiCl2paralleled the more potent effects of crystalline NiS in slowing and arresting cell proliferation. Crystalline NiS caused pronounced cell cycle specific blockage at a considerably lower concentration than was required for NiCl2to similarly arrest cell growth. Flow cytometry analysis showed that both compounds selectively blocked cell cycle progression in S phase (DNA synthetic stage). These results are suggestive of a common mechanism and site of toxicity inherent to both compounds and related to the cell growth phase during which DNA is replicated.