Synthetic 1,4-anthracenedione analogs induce cytochrome c release, caspase-9,-3, and-8 activities, poly(ADP-ribose) polymerase-1 cleavage and internucleosomal DNA fragmentation in HL-60 cells by a mechanism which involves caspase-2 activation but not Fas signaling
Synthetic 1,4-anthracenedione analogs induce cytochrome c release, caspase-9,-3, and-8 activities, poly(ADP-ribose) polymerase-1 cleavage and internucleosomal DNA fragmentation in HL-60 cells by a mechanism which involves caspase-2 activation but not Fas signaling
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DOI：
10.1016/j.bcp.2003.09.012
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发表时间：
2004-02-01
影响因子：
5.8
通讯作者：
Perchellet, JP
中科院分区：
文献类型：
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作者：
Perchellet, EM;Wang, Y;Perchellet, JP
对1，4-蒽醌（AQ代码号）的合成类似物（一种在体外模拟柔红霉素（道诺霉素）在纳摩尔范围内的抗增殖作用但具有阻断核苷转运并保留其在多药耐药肿瘤细胞中的功效的化合物）在HL-60细胞系统中诱导细胞凋亡的能力进行了测试。AQ 10，特别是新的先导抗增殖化合物AQ 8和AQ 9比AQ 1更有效地减少野生型、药物敏感性HL-60-S细胞的生长和完整性，这表明C6处的各种甲基取代基可以增强母体化合物的生物活性。核小体间DNA断裂，一种晚期凋亡标志物，类似地通过增加AQ 8和AQ 9的浓度在24 hr. Poly（ADP-核糖）聚合酶-1（PARP-1）裂解，一种细胞致力于凋亡所需的早期事件，在用AQ 9处理的HL-60-S细胞中在3-6 hr内检测到。与caspase 9和3级联负责PARP-1裂解的事实雅阁，在HL-60-S和多药耐药HL-60-RV细胞系中，AQ 9以相同的时间和浓度依赖性方式诱导起始caspase-9和效应caspase-3的活性，并达到相同的最大程度。有趣的是，1小时的脉冲处理足以使AQ 8和AQ 9在6小时时最大程度地诱导caspase-9和-3活性。在用AQ 9处理的HL-60-S细胞中，在3-6小时内也检测到线粒体细胞色素c（Cyt c）的释放，这一发现与Cyt c是激活caspase-9的凋亡触发剂的事实一致。此外，AQ类似物诱导与其降低肿瘤细胞生长和完整性的能力相关的Cyt c释放、半胱天冬酶-9和半胱天冬酶-3活性以及PARP-1裂解，AQ 8和AQ 9始终是最有效的。由于顶端半胱天冬酶2和8都可以作用于线粒体的上游以促进Cyt c释放，因此表明AQ 9分别在6小时和9小时最大程度地诱导半胱天冬酶-2和-8活性是有意义的。在AQ 8处理期间，半胱天冬酶-2抑制剂苄氧基羰基（z）-Val-Asp-Val-Ala-Asp（VDVAD）-氟甲基酮（falcine）完全阻断半胱天冬酶-9、-3和-8活化，而半胱天冬酶-8抑制剂z-Ile-Glu-Thr-Asp-（IETD）-falcine不阻止半胱天冬酶-2、-9和-3活化，表明AQ-induced caspase-2活性是下游caspase 9和3级联激活的关键上游事件，包括通过caspase-8的线粒体扩增环。然而，这些半胱天冬酶-2和-8抑制剂未能改变AQ 8诱导的Cyt c释放，表明AQs也可能独立于半胱天冬酶活化而靶向线粒体。此外，拮抗性抗Fas DX 2和ZB 4单克隆抗体（mAb）（其阻断激动性抗Fas CH 11 mAb诱导的Cyt c释放和半胱天冬酶-2、-8和-9活性）和中和性抗Fas配体（FasL）NOK-1 mAb均不能抑制AQ 9诱导的Cyt c释放和半胱天冬酶-2、-8和-9活性，提示FasL/Fas信号通路不参与抗增殖AQ类似物触发HL-60细胞凋亡的机制。(C)2003年爱思唯尔公司All rights reserved.
Synthetic analogs of 1,4-anthraquinone (AQ code number), a compound that mimics the antiproliferative effects of daunorubicin (daunomycin) in the nanomolar range in vitro but has the advantage of blocking nucleoside transport and retaining its efficacy in multidrug-resistant tumor cells, were tested for their ability to induce apoptosis in the HL-60 cell system. AQ10 and, especially, the new lead antiproliferative compounds AQ8 and AQ9 reduce the growth and integrity of wild-type, drug-sensitive, HL-60-S cells more effectively than AQ1, suggesting that various methyl group substituents at C6 may enhance the bioactivity of the parent compound. Internucleosomal DNA fragmentation, a late marker of apoptosis, is similarly induced in a biphasic manner by increasing concentrations of AQ8 and AQ9 at 24 hr. Poly(ADP-ribose) polymerase-1 (PARP-1) cleavage, an early event required for cells committed to apoptosis, is detected within 3-6 hr in HL-60-S cells treated with AQ9. In accord with the fact that the caspases 9 and 3 cascade is responsible for PARP-1 cleavage, the activities of initiator caspase-9 and effector caspase-3 are induced by AQ9 in the same time- and concentration-dependent manners and to the same maximal degrees in both the HL-60-S and multidrug-resistant HL-60-RV cell lines. Interestingly, a 1-hr pulse treatment is sufficient for AQ8 and AQ9 to maximally induce caspase-9 and -3 activities at 6 hr. The release of mitochondrial cytochrome c (Cyt c) is also detected within 3-6 hr in HL-60-S cells treated with AQ9, a finding consistent with the fact that Cyt c is the apoptotic trigger that activates caspase-9. Moreover, AQ analogs induce Cyt c release, caspase-9 and -3 activities and PARP-1 cleavage in relation with their abilities to decrease tumor cell growth and integrity, AQ8 and AQ9 being consistently the most effective. Since apical caspases 2 and 8 may both act upstream of mitochondria to promote Cyt c release, it is significant to show that AQ9 maximally induces caspase-2 and -8 activities at 6 and 9 hr, respectively. During AQ8 treatment, the caspase-2 inhibitor benzyloxycarbonyl (z)-Val-Asp-Val-Ala-Asp (VDVAD)-fluoromethyl ketone (fmk) totally blocks caspase-9, -3, and -8 activations, whereas the caspase-8 inhibitor z-Ile-Glu-Thr-Asp-(IETD)-fmk does not prevent caspase-2, -9, and -3 activations, suggesting that AQ-induced caspase-2 activity is an upstream event critical for the activation of the downstream caspases 9 and 3 cascade, including the mitochondrial amplification loop through caspase-8. However, these caspase-2 and -8 inhibitors fail to alter AQ8-induced Cyt c release, suggesting that AQs might also target mitochondria independently from caspase activation. Furthermore, the antagonistic anti-Fas DX2 and ZB4 monoclonal antibodies (mAbs), which block the induction of Cyt c release and caspase-2, -8, and -9 activities by the agonistic anti-Fas CH11 mAb, and the neutralizing anti-Fas ligand (FasL) NOK-1 mAb all fail to inhibit AQ9-induced Cyt c release and caspase-2, -8, and -9 activities,suggesting that the FasL/Fas signaling pathway is not involved in the mechanism by which antiproliferative AQ analogs trigger apoptosis in HL-60 cells. (C) 2003 Elsevier Inc. All rights reserved.