Cancer cell mitochondria are direct proapoptotic targets for the marine antitumor drug lamellarin D
Cancer cell mitochondria are direct proapoptotic targets for the marine antitumor drug lamellarin D
复制标题
DOI：
10.1158/0008-5472.can-05-1929
复制
发表时间：
2006-03-15
期刊：
影响因子：
11.2
通讯作者：
Bailly, C
中科院分区：
文献类型：
--
作者：
Kluza, J;Gallego, MA;Bailly, C
片螺素D是一种海洋生物碱，对一大组癌细胞系具有明显的细胞毒性，是拓扑异构酶I的有效抑制剂。然而，片螺素D对耐参照拓扑异构酶I毒物喜树碱的细胞系保持显著的细胞毒性。因此，我们假设拓扑异构酶I不是药物的唯一细胞靶点。使用互补的基于细胞的分析，我们提供的证据表明，片层蛋白D作用于癌细胞线粒体诱导凋亡。与喜树碱不同，片螺素D在P388白血病细胞系中诱导内部线粒体跨膜电位（Δ psi（m））的早期破坏。这种功能改变在很大程度上被环孢菌素A（一种线粒体通透性转换（MPT）的抑制剂）阻止，但不被半胱天冬酶抑制剂阻止。苄氧基羰基-Val-Ala-Asp（Ome）-氟甲基酮。Δ psi（m）破坏与线粒体肿胀和细胞色素c渗漏相关。使用一个可靠的实时流式细胞仪监测Δ psi（m），和肿胀的线粒体分离自白血病细胞，我们表明，片螺素D有一个直接的NIPT诱导作用。此外，在无细胞系统中需要线粒体来介导片层蛋白D诱导的核凋亡。片螺素D的直接线粒体效应解释了拓扑异构酶I突变的P388 CPT 5细胞对喜树碱耐药的敏感性。有趣的是，一个肿瘤活性类似物的lamellarin D，指定PM 031379，也发挥了直接的促凋亡作用的线粒体，与非肿瘤细胞系相比，对肿瘤细胞系的线粒体更显着的活动。总而言之，这项工作加强了片层素的药理学意义，并将片层素D定义为寻找抗化疗耐药癌细胞治疗方法的领导者。
Lamellarin D is a marine alkaloid with a pronounced cytotoxicity against a large panel of cancer cell lines and is a potent inhibitor of topoisomerase I. However, lamellarin D maintains a marked cytotoxicity toward cell lines resistant to the reference topoisomerase I poison camptothecin. We therefore hypothesized that topoisomerase I is not the only cellular target for the drug. Using complementary cell-based assays, we provide evidence that lamellarin D acts on cancer cell mitochondria to induce apoptosis. Lamellarin D, unlike camptothecin, induces early disruption of the inner mitochondrial transmembrane potential (Delta psi(m)) in the P388 leukemia cell line. The functional alterations are largely prevented by cyclosporin A, an inhibitor of the mitochondrial permeability transition (MPT), but not by the inhibitor of caspases.. benzyloxycarbonyl-Val-Ala-Asp(Ome)-fluoromethylketone. Delta psi(m) disruption is associated with mitochondrial swelling and cytochrome c leakage. Using a reliable real-time flow cytometric monitoring of Delta psi(m), and swelling of mitochondria isolated from leukemia cells, we show that lamellarin D has a direct NIPT-inducing effect. Furthermore, mitochondria are required in a cell-free system to mediate lamellarin D-induced nuclear apoptosis. The direct mitochondrial effect of lamellarin D accounts for the sensitivity of topoisomerase I-mutated P388CPT5 cells resistant to camptothecin. Interestingly, a tumor-active analogue of lamellarin D, designated PM031379, also exerts a direct proapoptotic action on mitochondria, with a more pronounced activity toward mitochondria of tumor cell lines compared with nontumor cell lines. Altogether, this work reinforces the pharmacologic interest of the lamellarins and defines lamellarin D as a lead in the search for treatments against chemoresistant cancer cells.