The cell cycle related differences in susceptibility of HL-60 cells to apoptosis induced by various antitumor agents.
The cell cycle related differences in susceptibility of HL-60 cells to apoptosis induced by various antitumor agents.
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发表时间：
1993-07
期刊：
影响因子：
11.2
通讯作者：
W. Gorczyca;J. Gong;B. Ardelt;F. Traganos;Z. Darżynkiewicz
中科院分区：
文献类型：
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作者：
W. Gorczyca;J. Gong;B. Ardelt;F. Traganos;Z. Darżynkiewicz
本研究旨在检测HL-60细胞对不同药物诱导的凋亡敏感性的细胞周期相关差异。用DNA拓扑异构酶I抑制剂喜树碱处理指数生长的HL-60细胞；DNA拓扑异构酶II抑制剂天尼泊苷、m-AMSA、米托蒽醌或Fostriecin；假定的酪氨酸激酶抑制剂染料木黄酮；丝氨酸/苏氨酸激酶抑制剂H7；蛋白质合成抑制剂环己亚胺；DNA复制抑制剂羟基脲；核苷类抗代谢物1- β - d -阿拉伯糖醛基胞嘧啶和5-氮杂胞苷；还有烷基化剂氮芥、顺铂、热疗和γ射线照射。通过两种不同的流式细胞术方法对这些药物诱导的细胞凋亡伴随的核内溶解进行了评估，一种是基于提取低分子量DNA后的DNA含量测量，另一种是使用外源性末端脱氧核苷酸转移酶来标记原位DNA链断裂。每种方法都允许鉴定凋亡细胞和分析未受影响细胞群的细胞周期分布；使用末端转移酶的方法还可以确定凋亡细胞的细胞周期位置。凝胶电泳DNA降解分析和细胞形态变化证实，大多数药物在给药后3小时就可观察到细胞凋亡，有些药物具有细胞周期特异性。通过S期的细胞在喜树碱、天尼坡苷、m-AMSA、米托蒽醌、H7、羟基脲和1- β - d -阿拉伯糖醛酸胞嘧啶处理时选择性敏感。G2-M细胞在H7或γ辐照下优先发生凋亡。G1期细胞优先受到5-氮杂胞苷、氮芥和热疗的影响。在Fostriecin、染料木素、环己亚胺或顺铂的病例中，没有观察到明显的细胞周期特异性。细胞周期相关的细胞凋亡易感性差异可能反映了给定药物引起的病变的严重程度和细胞修复病变的能力；两者都可以根据细胞周期阶段而变化。
The studies were aimed to detect the cell cycle-associated differences in the susceptibility of HL-60 cells to apoptosis induced by diverse agents. Exponentially growing HL-60 cells were treated with the DNA topoisomerase I inhibitor camptothecin; the DNA topoisomerase II inhibitors teniposide, m-AMSA, Mitoxantrone, or Fostriecin; the presumed tyrosine kinase inhibitor genistein; a serine/threonine kinase inhibitor H7; the protein synthesis inhibitor cycloheximide; the DNA replication inhibitor hydroxyurea; the nucleoside antimetabolites 1-beta-D-arabinofuranosylcytosine and 5-azacytidine; and the alkylating agent nitrogen mustard, cisplatin, hyperthermia, and gamma irradiation. Endonucleolysis, which accompanied apoptosis induced by these agents, was assessed by two different flow cytometric methods, one based on DNA content measurements following extraction of low molecular weight DNA, and another using exogenous terminal deoxynucleotidyl transferase to label in situ DNA strand breaks. Each method allowed for both identification of apoptotic cells and analysis of the cell cycle distribution of the unaffected cell population; the method using terminal transferase also allowed for identification of the cell cycle position of apoptotic cells. Confirmed by analysis of DNA degradation by gel electrophoresis and changes in cell morphology, apoptosis was observed as early as 3 h after administration of most drugs and for some drugs was cell cycle phase specific. Cells progressing through S phase were selectively susceptible when treated with camptothecin, teniposide, m-AMSA, Mitoxantrone, H7, hydroxyurea, and 1-beta-D-arabinofuranosylcytosine. Cells in G2-M preferentially underwent apoptosis in cultures treated with H7 or with gamma-irradiation. Cells in G1 phase were preferentially affected by 5-azacytidine, nitrogen mustard, and hyperthermia. No significant cell cycle specificity was observed in the case of Fostriecin, genistein, cycloheximide, or cisplatin. The cell cycle related difference in susceptibility to apoptosis may be a reflection of both the severity of the lesion induced by a given drug and the ability of the cells to repair that lesion; both can vary depending on the cell cycle phase.