Inhibition of autophagy with chloroquine is effective in melanoma
Inhibition of autophagy with chloroquine is effective in melanoma
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DOI：
10.1016/j.jss.2013.04.055
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发表时间：
2013-09-01
影响因子：
2.2
通讯作者：
McNally, Lacey R.
中科院分区：
文献类型：
--
作者：
Egger, Michael E.;Huang, Justin S.;McNally, Lacey R.
背景：癌细胞通过激活自噬途径来适应细胞生长加速和营养缺乏所产生的压力。缺氧诱导因子 1 α (HIF-1 α) 和热休克蛋白 90 (Hsp 90) 是两种允许细胞在代谢应激和缺氧情况下生长的蛋白质。我们假设氯喹 (CQ)（一种抑制自噬体功能的抗疟药）与棘霉素（一种 HIF-1 α 抑制剂）或 17-二甲氨基乙基氨基-17-二甲氧基格尔德霉素 (17-DMAG)（一种 Hsp 90 抑制剂）联合使用，将导致黑色素瘤的细胞毒性。 方法：使用 CQ 与棘霉素或 17-DMAG 组合对多种人类黑色素瘤细胞系（BRAF 野生型和突变型）进行体外测试。这些治疗在低氧 (5% O-2) 和常氧 (18% O-2) 条件下进行。通过自噬相关蛋白 HIF-1 α 和 Hsp 90 的蛋白质印迹确定作用机制。结果：氯喹、棘霉素和 17-DMAG 在常氧和缺氧条件下均能诱导多种人黑色素瘤细胞系的细胞毒性。氯喹与棘霉素联合在多种黑色素瘤细胞系（BRAF野生型和突变型）中在缺氧条件下实现了协同细胞毒性。蛋白质印迹分析表明，棘霉素单独使用以及与 CQ 联合使用均可降低 HIF-1 α 水平。 LC3通量的变化表明CQ疗法在自噬体水平上抑制了自噬。结论：用抗疟药CQ靶向自噬可能是黑色素瘤的有效癌症疗法。对氯喹的敏感性与 BRAF 突变状态无关。将 CQ 与 HIF-1 α 抑制剂棘霉素结合可改善缺氧条件下的细胞毒性。 (C) 2013 年，爱思唯尔公司出版。
Background: Cancer cells adapt to the stress resulting from accelerated cell growth and a lack of nutrients by activation of the autophagy pathway. Two proteins that allow cell growth in the face of metabolic stress and hypoxia are hypoxia-inducible factor-1 alpha (HIF-1 alpha) and heat shock protein 90 (Hsp 90). We hypothesize that chloroquine (CQ), an antimalarial drug that inhibits autophagosome function, in combination with either echinomycin, a HIF-1 alpha inhibitor, or 17-dimethylaminoethylamino-17-dimethoxygeldanamycin (17-DMAG), an Hsp 90 inhibitor, will result in cytotoxicity in melanoma.Materials and methods: Multiple human melanoma cell lines (BRAF wild-type and mutant) were tested in vitro with CQ in combination with echinomycin or 17-DMAG. These treatments were performed in hypoxic (5% O-2) and normoxic (18% O-2) conditions. Mechanism of action was determined through Western blot of autophagy-associated proteins HIF-1 alpha and Hsp 90.Results: Chloroquine, echinomycin, and 17-DMAG each induced cytotoxicity in multiple human melanoma cell lines, in both normoxia and hypoxia. Chloroquine combined with echinomycin achieved synergistic cytotoxicity under hypoxic conditions in multiple melanoma cell lines (BRAF wild-type and mutant). Western blot analysis indicated that echinomycin reduced HIF-1 alpha levels, both alone and in combination with CQ. Changes in LC3 flux indicated inhibition of autophagy at the level of the autophagosome by CQ therapy.Conclusions: Targeting autophagy with the antimalarial drug CQ may be an effective cancer therapy in melanoma. Sensitivity to chloroquine is independent of BRAF mutational status. Combining CQ with the HIF-1 alpha inhibitor echinomycin improves cytotoxicity in hypoxic conditions. (C) 2013 Published by Elsevier Inc.