Different sensitivity of chromatin to acid denaturation in quiescent and cycling cells as revealed by flow cytometry.
Different sensitivity of chromatin to acid denaturation in quiescent and cycling cells as revealed by flow cytometry.
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流式细胞术显示静止细胞和循环细胞中染色质对酸变性的不同敏感性。
DOI：
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发表时间：
1979
影响因子：
3.2
通讯作者：
M. Melamed
中科院分区：
文献类型：
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作者：
Z. Darżynkiewicz;F. Traganos;M. Andreeff;T. Sharpless;M. Melamed
用吖啶橙子染色所反映的原位DNA的性质在静止的未受刺激的淋巴细胞中与在有丝分裂原刺激后进入细胞周期的间期淋巴细胞相比是不同的。在用pH 1.5（1.3-1.9范围）的缓冲液处理细胞，然后用pH 2.6（2.3-2.9）的吖啶橙子染色后，观察到差异。在这些条件下，吖啶橙-DNA复合物的红色异染荧光在静止细胞中比在循环淋巴细胞中更高，而正染绿色荧光在循环间期细胞中更高。结果表明，在静止的淋巴细胞的凝聚染色质中的DNA（如在中期染色体）是更敏感的酸变性比分散的染色质中的DNA的周期间期细胞。该现象用于G 0和G1细胞之间以及G2和M细胞之间的流式细胞术区分。与正常淋巴细胞相反，应用于肿瘤细胞的方法表明存在染色质浓缩的细胞亚群，但DNA含量不仅是G1细胞的特征，也是S和G2细胞的特征。这些细胞代表静止（休息）亚群，在G1，S和/或G2逮捕的可能性进行了讨论。
The properties of DNA in situ as reflected by its staining with acridine orange are different in quiescent nonstimulated lymphocytes as compared with interphase lymphocytes that have entered the cell cycle after stimulation by mitogens. The difference is seen after cell treatment with buffers at pH 1.5 (1.3-1.9 range) followed by staining with acridine orange at pH 2.6 (2.3-2.9). Under these conditions the red metachromatic fluorescence of the acridine orange-DNA complex is higher in quiescent cells than in the cycling lymphocytes while the orthochromatic green fluorescence is higher in the cycling, interphase cells. The results suggest that DNA in condensed chromatin of quiescent lymphocytes (as in metaphase chromosomes) is more sensitive to acid-denaturation than DNA in dispersed chromatin of the cycling interphase cells. The phenomenon is used for flow cytometric differentiation between G0 and G1 cells and between G2 and M cells. In contrast to normal lymphocytes the method applied to neoplastic cells indicates the presence of cell subpopulations with condensed chromatin but with DNA content characteristic not only of G1 but also of S and G2 cells. The possibility that these cells represent quiescent (resting) subpopulations, arrested in G1, S and/or G2, is discussed.