Absence of RIPK3 predicts necroptosis resistance in malignant melanoma.
Absence of RIPK3 predicts necroptosis resistance in malignant melanoma.
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DOI：
10.1038/cddis.2015.240
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发表时间：
2015-09-10
影响因子：
9
通讯作者：
Leverkus M
中科院分区：
文献类型：
--
作者：
Geserick P;Wang J;Schilling R;Horn S;Harris PA;Bertin J;Gough PJ;Feoktistova M;Leverkus M
对凋亡和坏死刺激的获得性或内在抗性被认为是恶性黑色素瘤治疗成功的主要障碍。凋亡抑制蛋白（IAPs）是多种细胞死亡信号平台介导的细胞凋亡和坏死细胞死亡的重要调节因子。在本报告中，我们研究了iap对恶性黑色素瘤细胞死亡调控的影响。IAPs的抑制使一组黑色素瘤细胞对死亡配体诱导的细胞死亡强烈敏感，令人惊讶的是，这种死亡主要是由细胞凋亡介导的，因为添加caspase抑制剂可以完全挽救细胞凋亡。有趣的是，坏死坏死信号的缺失与所有细胞系中受体相互作用蛋白激酶3 (RIPK3) mRNA和蛋白表达的缺乏相关，而原代黑素细胞和培养的痣细胞强烈表达RIPK3。在一组黑色素瘤细胞系中重构RIPK3，而不是RIPK3激酶死亡突变体，克服了CD95L/IAP拮抗剂诱导的不依赖自分泌肿瘤坏死因子分泌的坏死性下垂抵抗。使用特异性抑制剂，功能研究表明，ripk3介导的混合谱系激酶结构域样蛋白（MLKL）磷酸化和necroptosis诱导非常需要受体相互作用蛋白激酶-1信号传导。此外，突变BRAF的抑制剂Dabrafenib，而不是Vemurafenib，在RIPK3存在时抑制黑色素瘤细胞的坏死。我们的数据表明，黑色素瘤中RIPK3的缺失和BRAF抑制剂Dabrafenib（而非Vemurafenib）对RIPK3/MLKL轴的选择性抑制是防止坏死性坏死的关键。允许RIPK3表达的策略可能允许揭示黑色素瘤中的坏死性坏死信号机制，并指出该途径的重新激活作为转移性黑色素瘤的治疗选择。
Acquired or intrinsic resistance to apoptotic and necroptotic stimuli is considered a major hindrance of therapeutic success in malignant melanoma. Inhibitor of apoptosis proteins (IAPs) are important regulators of apoptotic and necroptotic cell death mediated by numerous cell death signalling platforms. In this report we investigated the impact of IAPs for cell death regulation in malignant melanoma. Suppression of IAPs strongly sensitized a panel of melanoma cells to death ligand-induced cell death, which, surprisingly, was largely mediated by apoptosis, as it was completely rescued by addition of caspase inhibitors. Interestingly, the absence of necroptosis signalling correlated with a lack of receptor-interacting protein kinase-3 (RIPK3) mRNA and protein expression in all cell lines, whereas primary melanocytes and cultured nevus cells strongly expressed RIPK3. Reconstitution of RIPK3, but not a RIPK3-kinase dead mutant in a set of melanoma cell lines overcame CD95L/IAP antagonist-induced necroptosis resistance independent of autocrine tumour necrosis factor secretion. Using specific inhibitors, functional studies revealed that RIPK3-mediated mixed-lineage kinase domain-like protein (MLKL) phosphorylation and necroptosis induction critically required receptor-interacting protein kinase-1 signalling. Furthermore, the inhibitor of mutant BRAF Dabrafenib, but not Vemurafenib, inhibited necroptosis in melanoma cells whenever RIPK3 is present. Our data suggest that loss of RIPK3 in melanoma and selective inhibition of the RIPK3/MLKL axis by BRAF inhibitor Dabrafenib, but not Vemurafenib, is critical to protect from necroptosis. Strategies that allow RIPK3 expression may allow unmasking the necroptotic signalling machinery in melanoma and points to reactivation of this pathway as a treatment option for metastatic melanoma.