Mechanism of cancer cell death induced by depletion of an essential replication regulator.
Mechanism of cancer cell death induced by depletion of an essential replication regulator.
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DOI：
10.1371/journal.pone.0036372
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Masai H
中科院分区：
文献类型：
--
作者：
Ito S;Ishii A;Kakusho N;Taniyama C;Yamazaki S;Fukatsu R;Sakaue-Sawano A;Miyawaki A;Masai H
复制因子的耗尽通常会导致癌细胞死亡。CDC7是一种启动DNA复制所必需的激酶，它的缺失会导致癌细胞死亡，而与其P53状态无关，但诱导细胞死亡的确切途径尚未确定。我们使用了最近发展起来的细胞周期指示剂Fucci，来精确地描述由CdC7耗尽诱导的细胞死亡过程。我们还通过与其他细胞周期调节因子的融合，产生并使用了类似的荧光细胞周期指示剂来分析P53阳性和阴性背景下活细胞的细胞死亡模式。我们发现，在P53阳性和阴性细胞中，CDC7耗尽可以诱导不同的细胞周期反应。P53阴性细胞主要暂时停滞在G2期，积聚细胞周期蛋白B1和其他有丝分裂调节因子。在细胞周期蛋白B_1积聚的情况下，细胞长时间停滞在G2期，突然进入异常的M期，细胞在有丝分裂后死亡。取消胞质CyclinB1积聚可部分减少细胞死亡。ATR-MK2途径负责14-3-3σ蛋白与细胞周期蛋白B1的结合。相比之下，P53阳性的癌细胞并不积聚细胞周期蛋白B1，但在Cdc7耗尽后，似乎主要通过进入异常的S期而死亡。CDC7抑制剂与已知抗癌药物的结合以一种基因依赖的方式显著刺激癌细胞的细胞死亡效应，为未来的联合治疗提供了战略基础。我们的结果表明，使用Fucci和类似的荧光细胞周期指示剂，提供了一种方便的检测系统，用于识别与癌细胞死亡相关的细胞周期事件。它们还表明，由于癌细胞中DNA复制的启动不足而导致的基因特异性细胞死亡模式及其开发有效的癌症治疗方法的潜力。
Depletion of replication factors often causes cell death in cancer cells. Depletion of Cdc7, a kinase essential for initiation of DNA replication, induces cancer cell death regardless of its p53 status, but the precise pathways of cell death induction have not been characterized. We have used the recently-developed cell cycle indicator, Fucci, to precisely characterize the cell death process induced by Cdc7 depletion. We have also generated and utilized similar fluorescent cell cycle indicators using fusion with other cell cycle regulators to analyze modes of cell death in live cells in both p53-positive and -negative backgrounds. We show that distinct cell-cycle responses are induced in p53-positive and -negative cells by Cdc7 depletion. p53-negative cells predominantly arrest temporally in G2-phase, accumulating CyclinB1 and other mitotic regulators. Prolonged arrest at G2-phase and abrupt entry into aberrant M-phase in the presence of accumulated CyclinB1 are followed by cell death at the post-mitotic state. Abrogation of cytoplasmic CyclinB1 accumulation partially decreases cell death. The ATR-MK2 pathway is responsible for sequestration of CyclinB1 with 14-3-3σ protein. In contrast, p53-positive cancer cells do not accumulate CyclinB1, but appear to die mostly through entry into aberrant S-phase after Cdc7 depletion. The combination of Cdc7 inhibition with known anti-cancer agents significantly stimulates cell death effects in cancer cells in a genotype-dependent manner, providing a strategic basis for future combination therapies. Our results show that the use of Fucci, and similar fluorescent cell cycle indicators, offers a convenient assay system with which to identify cell cycle events associated with cancer cell death. They also indicate genotype-specific cell death modes induced by deficient initiation of DNA replication in cancer cells and its potential exploitation for development of efficient cancer therapies.
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DOI：
10.1083/jcb.115.1.1
发表时间：
1991-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Pines J;Hunter T
通讯作者：
Hunter T
影响因子：
7.8
作者：
Jin, P;Hardy, S;Morgan, DO
通讯作者：
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影响因子：
21.3
作者：
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通讯作者：
Medema, RH
影响因子：
11.4
作者：
Kim, JM;Nakao, K;Masai, H
通讯作者：
Masai, H
影响因子：
4.3
作者：
Matsumoto, Seiji;Shimmoto, Michie;Masai, Hisao
通讯作者：
Masai, Hisao