Alsterpaullone, a novel cyclin-dependent kinase inhibitor, induces apoptosis by activation of caspase-9 due to perturbation in mitochondrial membrane potential
Alsterpaullone, a novel cyclin-dependent kinase inhibitor, induces apoptosis by activation of caspase-9 due to perturbation in mitochondrial membrane potential
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DOI：
10.1002/mc.10114
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发表时间：
2003-04-01
影响因子：
4.6
通讯作者：
Senderowicz, AM
中科院分区：
文献类型：
--
作者：
Lahusen, T;De Siervi, A;Senderowicz, AM
由于细胞周期蛋白依赖性激酶(CDK)的过度激活，大多数人类肿瘤都存在视网膜母细胞瘤通路的异常。基于这一观察，正在开发新的小分子，如黄烷醇和UCN-01，目前正在进行临床测试。开发CDK调节剂的努力使我们发现了一类新的CDK抑制剂，即Paullone[癌症研究报告1999；59：2566]。初步研究表明，白藜芦酮在体外抑制CDKs，从而阻止细胞周期进展。然而，马兜铃内酯抗增殖作用的确切机制从未被探索过。在这份报告中，我们首次证明了最有效的泡桐酮(ALP)诱导Jurkat细胞系凋亡并促进克隆形成能力的丧失。碱性磷酸酶导致caspase-8和-9的早期激活，导致caspase-3和多聚(ADP-核糖)聚合酶(PARP)的裂解。此外，碱性磷酸酶诱导的细胞凋亡与XIAP或BCL-XL等抗凋亡蛋白的缺失无关。细胞通透性抑制剂z-Asp(Om)-Glu(Om)-Val-Asp(Ome)-氟甲基酮和苯氧甲酰-Val-Ala-Asp(Om)-氟甲基酮(ZVAD)预先孵育可阻断ALP诱导的细胞凋亡。此外，一般的caspase抑制剂zVAD阻断了除caspase-9以外的大多数caspase的切割和激活。线粒体膜电位的研究也表明，在zVAD存在的情况下，ALP能够破坏线粒体的电位，这表明ALP激活caspase-9是在线粒体扰动之后进行的。ZVAD预先孵育Jurkat细胞并不能阻止细胞周期蛋白D3的耗竭、CDK的丢失或ALP对细胞周期的抑制。综上所述，这些实验表明，ALP通过线粒体扰动激活caspase-9。激活的caspase-9裂解并激活caspase-8和caspase-3，导致细胞凋亡。在一般caspase抑制剂zVAD存在下，ALP对细胞周期的影响没有改变，而细胞凋亡被阻断，这表明ALP的CDK效应不足以诱导ALP诱导的细胞凋亡。有必要对保乐酮进行更多的研究，以进一步确定它们的临床前效应，并探索它们在临床环境中的潜在用途。2003年出版，Wiley-Liss，Inc.(Dagger)。
The majority of human neoplasms have aberrations in the retinoblastoma pathway due to hyperactivation of cyclin-dependent kinases (CDK). Based on this observation, novel small molecules, such as flavopiridol and UCN-01, are being developed and are currently being tested in the clinic. Efforts to develop CDK modulators led us to the discovery of a novel class of CDK inhibitors, the paullones [Cancer Res 1999;59:2566]. initial studies demonstrated that paullones inhibit CDKs in vitro, thereby blocking cell-cycle progression. However, the exact mechanism for the antiproliferative effects of paullones was never explored. In this report, we demonstrate for the first time that the most potent paullone, alsterpaullone (Alp), induced apoptosis and promoted loss in clonogenicity in the Jurkat cell line. Alp caused early activation of both caspase-8 and -9, leading to cleavage of caspase-3 and poly(ADP-ribose) polymerase (PARP). Moreover, apoptosis by Alp was not associated with loss in anti-apoptotic proteins such as XIAP or BCL-XL. Pre-incubation with cell-permeable inhibitors z-Asp(OMe)-Glu(OMe)-Val-Asp(Ome)-fluoromethyl ketone and benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (ZVAD) blocked Alp-induced apoptosis. Moreover, the general caspase inhibitor ZVAD blocked the cleavage and activation of most caspases tested except caspase-9. Studies of mitochondrial membrane potential also demonstrated that Alp is able to disrupt mitochondrial potential in the presence of ZVAD, suggesting that the activation of caspase-9 by Alp follows mitochondrial perturbation. Preincubation of Jurkat cells with ZVAD did not prevent the depletion of cyclin D3, loss of CDK, or cell-cycle arrest by Alp. In summary, these experiments suggest that Alp activates caspase-9 via mitochondrial perturbation. Active caspase-9 cleaves and activates caspase-8 and caspase-3, leading to apoptosis. In the presence of the general caspase inhibitor ZVAD, the cell-cycle effects of Alp are unaltered while apoptosis is blocked, suggesting that the CDK effects of Alp are not sufficient for Alp-induced apoptosis. Additional studies with paullones are warranted to further characterize their preclinical effects and to explore their potential use in the clinical setting. Published 2003 Wiley-Liss, Inc.(dagger).