E2F1-dependent pathways are involved in amonafide analogue 7-d-induced DNA damage, G2/M arrest, and apoptosis in p53-deficient K562 cells
E2F1-dependent pathways are involved in amonafide analogue 7-d-induced DNA damage, G2/M arrest, and apoptosis in p53-deficient K562 cells
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DOI：
10.1002/jcb.24194
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发表时间：
2012-10-01
影响因子：
4
通讯作者：
Qian, Xuhong
中科院分区：
文献类型：
--
作者：
Li, Yiquan;Shao, Jin;Qian, Xuhong
众所周知，E2 F1基因在调节从G1进入S期中起关键作用，而涉及它的补救抗肿瘤途径，特别是在p53缺失的情况下，尚未完全表征。因此，我们试图确定参与E2 F1依赖性促凋亡途径激活的上游和下游事件。为此目的，筛选了氨萘非特类似物7-d（2-（3-（2-（二甲基氨基）乙氨基）丙基）-6-（十二烷氨基）-1H-苯并[de]异喹啉-1，3（2 H）-二酮），其对p53缺陷型人慢性髓性白血病（CML）K562细胞表现出高的抗肿瘤活性。流式细胞术和Western blot分析显示，7-d处理的K562细胞明显的G2/M周期阻滞和凋亡，并呈剂量和时间依赖性，通过p53非依赖性途径。彗星尾部形成的显著增加，H2 AX的表达表明，DNA双链断裂（DSB）造成的处理7天。据报道，ATM/ATR信号转导将E2 F1诱导与响应于DNA损伤的凋亡联系起来。ATM/ATR信号通路抑制剂咖啡因可拮抗7-d诱导的G2/M期阻滞和细胞凋亡，表明ATM/ATR信号通路被7-d处理激活。此外，E2 F1，p73，Apaf-1和p73从HDM 2解离的表达增加诱导7-d的治疗，然而，E2 F1表达的敲除逆转p73，Apaf-1，和p21 Cip 1/WAF 1的表达，重新激活细胞周期进程，并抑制7-d诱导的凋亡。总之，我们的研究结果首次表明，7-d介导其对CML p53缺陷细胞的生长抑制作用，通过激活E2 F1依赖性线粒体和细胞周期检查点信号通路，随后靶向p73，Apaf-1和p21 Cip 1/WAF 1。J.细胞。113：31653177，2012。(c)2012年威利期刊公司
The E2F1 gene well known is its pivotal role in regulating the entry from G1 to S phase, while the salvage antitumoral pathway which implicates it, especially in the absence of p53, is not fully characterized. We therefore attempted to identify the up- and down-stream events involved in the activation of the E2F1-dependent pro-apoptotic pathway. For this purpose, a amonafide analogue, 7-d (2-(3-(2-(Dimethylamino)ethylamino)propyl)-6-(dodecylamino)-1H-benzo[de]isoquinoline-1,3(2H)-dione) was screened, which exhibited high antitumor activity against p53-deficient human Chronic Myelogenous Leukemia (CML) K562 cells. Analysis of flow cytometry and western blots of K562 cells treated with 7-d revealed an appreciable G2/M cycle arrest and apoptosis in a dose and time-dependent manner via p53-independent pathway. A striking increase in Comet tail formation and ?-H2AX expression showed that DNA double strand breaks (DSB) were caused by 7-d treatment. ATM/ATR signaling was reported to connect E2F1 induction with apoptosis in response to DNA damage. Indeed, 7-d-induced G2/M arrest and apoptosis were antagonized by ATM/ATR signaling inhibitor, Caffeine, which suggested that ATM/ATR signaling was activated by 7-d treatment. Furthermore, the increased expression of E2F1, p73, and Apaf-1 and p73 dissociation from HDM2 was induced by 7-d treatment, however, knockout of E2F1 expression reversed p73, Apaf-1, and p21Cip1/WAF1 expression, reactivated cell cycle progression, and inhibited 7-d-induced apoptosis. Altogether our results for the first time indicate that 7-d mediates its growth inhibitory effects on CML p53-deficient cells via the activation of an E2F1-dependent mitochondrial and cell cycle checkpoint signaling pathway which subsequently targets p73, Apaf-1, and p21Cip1/WAF1. J. Cell. Biochem. 113: 31653177, 2012. (c) 2012 Wiley Periodicals, Inc.