Centre for Immunobiology, The Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary London University, 4 Newark Street, London E1 2AT, UK.
Flow Cytometry Core Facility, The Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary London University, 4 Newark Street, London E1 2AT, UK.
Int J Mol Sci. 2019 Sep 6;20(18):4379. doi: 10.3390/ijms20184379.
The term necrosis is commonly applied to cells that have died via a non-specific pathway or mechanism but strictly is the description of the degradation processes involved once the plasma membrane of the cell has lost integrity. The signalling pathways potentially involved in accidental cell death (ACD) or oncosis are under-studied. In this study, the flow cytometric analysis of the intracellular antigens involved in regulated cell death (RCD) revealed the phenotypic nature of cells undergoing oncosis or necrosis. Sodium azide induced oncosis but also classic apoptosis, which was blocked by zVAD (z-Vla-Ala-Asp(OMe)-fluoromethylketone). Oncotic cells were found to be viability/caspase-3/RIP3 (Receptor-interacting serine/threonine protein kinase 3). These two cell populations also displayed a DNA damage response (DDR) phenotype pH2AX/PARP, cleaved PARP induced caspase independent apoptosis H2AX/PARP and hyper-activation or parthanatos H2AX/PARP. Oncotic cells with phenotype cell viability/RIP3/caspase-3 showed increased DDR and parthanatos. Necrostatin-1 down-regulated DDR in oncotic cells and increased sodium azide induced apoptosis. This flow cytometric approach to cell death research highlights the link between ACD and the RCD processes of programmed apoptosis and necrosis.
“坏死”一词通常用于描述通过非特异性途径或机制死亡的细胞，但严格来说，它是指细胞的质膜失去完整性后所涉及的降解过程。目前，关于潜在参与意外细胞死亡（ACD）或胀亡的信号通路研究还较少。在这项研究中，通过流式细胞术分析参与细胞程序性死亡（RCD）的细胞内抗原，揭示了发生胀亡或坏死的细胞的表型特征。叠氮化钠诱导胀亡，但也诱导经典凋亡，而 zVAD（z-Val-Ala-Asp(OMe)-氟甲基酮）可以阻断经典凋亡。发现胀亡细胞的活性/半胱天冬酶-3/RIP3（受体相互作用丝氨酸/苏氨酸蛋白激酶 3）。这两种细胞群也表现出 DNA 损伤反应（DDR）表型 pH2AX/PARP，caspase 非依赖性凋亡诱导的 PARP 片段 pH2AX/PARP，以及超激活或 parthanatos pH2AX/PARP。表现为细胞活性/RIP3/半胱天冬酶-3 的胀亡细胞显示出 DDR 和 parthanatos 的增加。坏死抑制蛋白-1 下调胀亡细胞中的 DDR，并增加叠氮化钠诱导的凋亡。这种细胞死亡研究的流式细胞术方法强调了 ACD 与程序性凋亡和坏死的 RCD 过程之间的联系。
