ACTIVATION OF PROGRAMMED CELL-DEATH (APOPTOSIS) BY CISPLATIN, OTHER ANTICANCER DRUGS, TOXINS AND HYPERTHERMIA
ACTIVATION OF PROGRAMMED CELL-DEATH (APOPTOSIS) BY CISPLATIN, OTHER ANTICANCER DRUGS, TOXINS AND HYPERTHERMIA
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DOI：
10.1016/0006-2952(90)90733-2
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发表时间：
1990-11-15
影响因子：
5.8
通讯作者：
EASTMAN, A
中科院分区：
文献类型：
--
作者：
BARRY, MA;BEHNKE, CA;EASTMAN, A
在中国仓鼠卵巢细胞系中研究了顺铂诱导的细胞死亡，其中一种细胞系具有 DNA 切除修复能力，另一种细胞系则缺乏（100 倍敏感）。先前的实验表明，细胞在死亡前会进展到细胞周期的 G2 期并停滞在其中。在 G2 停滞后和膜完整性丧失之前检测到 DNA 双链断裂。人们对这些 DNA 断裂进行了更详细的研究。观察到 DNA 片段由大约 180 个碱基对的多聚体组成。这些片段与核酸内切酶对染色质的核小体间切割一致。在 LC90 浓度下，顺铂处理后 48 小时开始 DNA 消化，24 小时后膜完整性丧失且细胞收缩。药物处理后 12 小时，高浓度的顺铂（170 个杀死日志）诱导 DNA 消化，但 12 小时后发生膜完整性丧失。放线菌酮抑制细胞死亡和 DNA 断裂，表明需要新的蛋白质合成。与许多其他试剂一起孵育的细胞表现出相同的 DNA 降解特征模式。在 90% 致死条件下，热疗可在 30 分钟内诱导 DNA 消化，甲氨蝶呤可在 18 小时内诱导 DNA 消化，所有其他测试试剂可在 48-72 小时内诱导 DNA 消化。 DNA 消化总是先于膜完整性丧失和细胞收缩。这些观察结果与细胞凋亡或程序性细胞死亡过程中发生的细胞死亡一致，并证明了 DNA 消化作为细胞死亡中早期且可能是必要步骤的重要性。结果表明，无论药物的主要作用位点如何，大多数药物引起的细胞死亡都是通过激活细胞凋亡信号转导途径介导的。结果还表明细胞凋亡有两种信号途径，一种与药物作用直接相关，另一种则需要细胞周期相关事件。
Cell death induced by cisplatin was studied in Chinese hamster ovary cell lines, one proficient and the other deficient (100-fold sensitive) in DNA excision repair. Previous experiments demonstrated that cells progressed to and arrested in the G2 phase of the cell cycle before dying. DNA double-strand breaks were detected following G2 arrest and prior to loss of membrane integrity. These DNA breaks have been studied in more detail. DNA fragments were observed consisting of multimers of approximately 180 base pairs. These fragments are consistent with internucleosomal cleavage of chromatin by an endonuclease. At LC90 concentrations, DNA digestion began 48 hr after cisplatin treatment followed by loss of membrane integrity and cell shrinkage 24 hr later. High concentrations of cisplatin (170 logs of kill) induced DNA digestion 12 hr after drug treatment but loss of membrane integrity occurred 12 hr later. Both cell death and DNA fragmentation were inhibited by cycloheximide, suggesting the requirement for new protein synthesis. Cells incubated with many other agents demonstrated the same characteristic pattern of DNA degradation. At 90% lethal conditions, DNA digestion was induced within 30 min by hyperthermia, 18 hr by methotrexate, and 48-72 hr by all other agents tested. DNA digestion always preceded loss of membrane integrity and cell shrinkage. These observations are consistent with cell death occurring by the process of apoptosis, or programmed cell death, and demonstrate the importance of DNA digestion as an early and presumably essential step in cell death. The results suggest that, irrespective of the primary site of action of a drug, cell death by most pharmacologic agents is mediated by activation of the signal transduction pathway for apoptosis. The results also suggest two signal pathways for apoptosis, one directly associated with drug action and a second that requires cell cycle-related events.