RIP3 Expression Induces a Death Profile Change in U2OS Osteosarcoma Cells After 5-ALA-PDT
RIP3 Expression Induces a Death Profile Change in U2OS Osteosarcoma Cells After 5-ALA-PDT
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DOI：
10.1002/lsm.21088
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发表时间：
2011-09-01
影响因子：
2.4
通讯作者：
Piette, Jacques
中科院分区：
文献类型：
--
作者：
Coupienne, Isabelle;Fettweis, Gregory;Piette, Jacques
背景和目的：受体相互作用蛋白3(RIP3)最近被认为是一种关键的坏死介质，但也被认为参与了细胞凋亡的调节。本研究的目的是比较5-氨基酮戊酸(5-ALA)介导光动力疗法(PDT)在RIP3基因缺失的U2OS细胞系和RIP3表达恢复的U2OS细胞中诱导的细胞死亡情况。用5-ALA-PDT处理野生型和RIP3-U2OS细胞。结果：RIP3-U2OS细胞的存活率明显高于野生型细胞。此外，zVAD-fmk可降低RIP3-U2OS细胞的死亡率。在这些细胞中，caspase-3、7、8、9和PARP的裂解也较高，表明caspase依赖的细胞凋亡被激活。同时，在RIP3-U2OS细胞中清楚地发现了自噬的推力。相反，RIP3-U2OS的坏死率低于野生型。结论：RIP3在U2OS细胞中的表达不仅能提高细胞的存活率，而且能改变细胞对光动力疗法的反应，使细胞死亡情况发生改变。与RIP3缺失的野生型细胞相比，凋亡和自噬途径明显上调。而在RIP3-U2OS细胞中，诱导的坏死作用较弱。在这种情况下，自噬很可能对PDT诱导的细胞死亡起到保护作用，并允许RIP3-U2OS细胞更好地存活。这项工作也强调了RIP3在凋亡途径中发挥的重要作用，尽管其方式仍然广泛未知。激光外科医生。2011年，43：557564。(C)2011年Wiley-Liss，Inc.
Background and Objective: The receptor-interacting protein 3 (RIP3) has recently been outlined as a key necrosis mediator but is also thought to participate in the regulation of apoptosis. The aim of this study is to compare the cell death profile induced by 5-aminolevulic acid (5-ALA)-mediated photodynamic therapy (PDT) in the RIP3-deficient cell line U2OS and in U2OS cells in which the expression of RIP3 was restored.Materials and Methods: RIP3-expressing U2OS cells (RIP3-U2OS) were obtained after transfection and antibiotic selection. Wild type and RIP3-U2OS cells were treated by 5-ALA-PDT. Overall cell viability was evaluated and different parameters characteristic of apoptosis, autophagy, and necrosis were studied.Results: Surprisingly, the survival of RIP3-U2OS cells was higher compared to that of the wild type cells. In addition, RIP3-U2OS cell death was decreased by a zVAD-fmk pre-treatment. A higher cleavage of caspase-3, 7, 8, 9, and PARP was also detected in these cells, pointing out to the activation of caspase-dependent apoptosis. In parallel, a thrust of autophagy was clearly identified in the RIP3-U2OS cells. Conversely, RIP3-U2OS exhibited a lower level of necrosis than the wild types. Interestingly, necrostatin-1 efficiently decreased necrosis level in RIP3-U2OS but not in wild type cells.Conclusion: Expression of RIP3 in U2OS cells led to a better survival but also to a death profile change in response to PDT. The apoptotic and autophagic pathways were clearly up-regulated compared to the RIP3-deficient wild type cells. However, induction of necrosis was weaker in the RIP3-U2OS cells. In this context, autophagy is likely to play a protective role against PDT-induced cell death and to allow a better survival of RIP3-U2OS cells. This work also highlights the important role played by RIP3 in the apoptotic pathway, although the modalities are still widely unknown. Lasers Surg. Med. 43: 557564, 2011. (C) 2011 Wiley-Liss, Inc.