Melo-Lima Sara, Celeste Lopes Maria, Mollinedo Faustino
Instituto de Biología Molecular y Celular del Cáncer, Centro de Investigación del Cáncer, CSIC-Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, Spain ; Centre for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
Centre for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal ; Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal.
Oncoscience. 2014 Oct 22;1(10):649-64. doi: 10.18632/oncoscience.89. eCollection 2014.
Necroptosis is a regulated necrotic cell death that involves receptor-interacting protein kinases RIPK1 and RIPK3. Here, we report that edelfosine triggers a rapid and massive cell death in human glioblastoma cells with characteristics of necrosis. Only a minor proportion of edelfosine-treated cells underwent caspase-dependent apoptosis. Autophagy and a rapid influx of extracellular calcium into the cells had little impact on cell death. Levels of procaspase-8 were very low in necroptosis-prone glioma cells compared with the levels in other cancer cell types that underwent apoptosis upon edelfosine treatment. The RIPK1-dependent necroptosis inhibitors necrostatin-1 (Nec-1) and Nec-1s as well as siRNA-mediated silencing of RIPK3 inhibited edelfosine-induced necroptosis, resulting in increased caspase-dependent apoptosis in edelfosine-treated glioblastoma U118 cells. Inhibition of the RIPK3 substrate MLKL with necrosulfonamide also increased apoptosis in edelfosine-treated cells. These data support a major role for RIPK1 and RIPK3 in the induction of necrotic cell death and in the switch from necrosis to apoptosis following edelfosine treatment. These results indicate that the ether lipid edelfosine exerts a rapid necroptotic cell death in apoptosis-reluctant glioblastoma cells, suggesting that induction of necroptosis could constitute a new approach for glioblastoma therapy.
坏死性凋亡是一种受调控的坏死性细胞死亡，涉及受体相互作用蛋白激酶RIPK1和RIPK3。在此，我们报告依地福新在人胶质母细胞瘤细胞中引发快速且大量的细胞死亡，具有坏死特征。仅一小部分经依地福新处理的细胞发生了半胱天冬酶依赖性凋亡。自噬以及细胞外钙快速流入细胞对细胞死亡影响很小。与经依地福新处理后发生凋亡的其他癌细胞类型相比，易发生坏死性凋亡的胶质瘤细胞中前半胱天冬酶-8水平非常低。RIPK1依赖性坏死性凋亡抑制剂坏死素-1（Nec-1）和Nec-1s以及siRNA介导的RIPK3沉默抑制了依地福新诱导的坏死性凋亡，导致经依地福新处理的胶质母细胞瘤U118细胞中半胱天冬酶依赖性凋亡增加。用坏死磺酰胺抑制RIPK3底物MLKL也增加了经依地福新处理的细胞中的凋亡。这些数据支持RIPK1和RIPK3在诱导坏死性细胞死亡以及依地福新处理后从坏死向凋亡转变中起主要作用。这些结果表明醚脂依地福新在抗凋亡的胶质母细胞瘤细胞中引发快速的坏死性凋亡细胞死亡，提示诱导坏死性凋亡可能构成胶质母细胞瘤治疗的一种新方法。
