Resistance to hypoxia-induced necroptosis is conferred by glycolytic pyruvate scavenging of mitochondrial superoxide in colorectal cancer cells.
Resistance to hypoxia-induced necroptosis is conferred by glycolytic pyruvate scavenging of mitochondrial superoxide in colorectal cancer cells.
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DOI：
10.1038/cddis.2013.149
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发表时间：
2013-05-02
影响因子：
9
通讯作者：
Yu, L. C. H.
中科院分区：
文献类型：
--
作者：
Huang, C-Y;Kuo, W-T;Huang, Y-C;Lee, T-C;Yu, L. C. H.
关键词：
癌细胞可以通过代谢重编程以实现高水平的无氧糖酵解，从而在氧气和营养匮乏的情况下生存，这有助于肿瘤生长和耐药性。异常表达的葡萄糖转运蛋白 (GLUT) 与人结直肠癌坏死周围区域的缺氧 (Hx) 诱导因子 (HIF)1α 共定位。然而，缺氧癌细胞中葡萄糖代谢赋予的抗坏死抵抗的潜在机制仍然知之甚少。我们的目的是研究 Hx 诱导的坏死性凋亡的信号通路，并探讨葡萄糖丙酮酸代谢物在死亡抵抗机制中的作用。将人结直肠癌细胞暴露于有或没有葡萄糖的 Hx 中，并通过受体相互作用蛋白 (RIP)1/3 激酶免疫沉淀和 32P 激酶测定来检查细胞坏死性凋亡。我们的结果显示，在无葡萄糖培养基中，缺氧细胞中 RIP1/3 复合物的形成和磷酸化增加，但正常氧细胞中则没有。通过 necrostatin-1 或基因沉默阻断 RIP1 信号传导，可减少乳脱氢酶 (LDH) 泄漏和质膜崩解。缺氧挑战后注意到线粒体超氧化物的产生；抗氧化剂的减少抑制了 RIP 信号传导和细胞坏死。补充葡萄糖可减少缺氧细胞中 RIP 依赖性 LDH 渗漏和形态损伤，而不可代谢的糖类似物则不会。给予葡萄糖的缺氧细胞表现出 HIF1α 的核转位，与 GLUT-1 和 GLUT-4 表达上调以及细胞内 ATP、丙酮酸和乳酸水平增加相关。葡萄糖介导的死亡抵抗被碘乙酸（3-磷酸甘油醛脱氢酶的抑制剂）消除，但被UK5099（线粒体丙酮酸载体的抑制剂）消除，这表明糖酵解途径参与了抗坏死机制。最后，用细胞可渗透的丙酮酸衍生物代替葡萄糖也可以通过以能量独立的方式抑制线粒体超氧化物来减少 Hx 诱导的坏死性凋亡。总之，糖酵解代谢部分通过线粒体自由基的丙酮酸清除，赋予缺氧癌细胞对 RIP 依赖性坏死性凋亡的抵抗力。
Cancer cells may survive under oxygen and nutrient deprivation by metabolic reprogramming for high levels of anaerobic glycolysis, which contributes to tumor growth and drug resistance. Abnormally expressed glucose transporters (GLUTs) are colocalized with hypoxia (Hx) inducible factor (HIF)1α in peri-necrotic regions in human colorectal carcinoma. However, the underlying mechanisms of anti-necrotic resistance conferred by glucose metabolism in hypoxic cancer cells remain poorly understood. Our aim was to investigate signaling pathways of Hx-induced necroptosis and explore the role of glucose pyruvate metabolite in mechanisms of death resistance. Human colorectal carcinoma cells were Hx exposed with or without glucose, and cell necroptosis was examined by receptor-interacting protein (RIP)1/3 kinase immunoprecipitation and 32P kinase assays. Our results showed increased RIP1/3 complex formation and phosphorylation in hypoxic, but not normoxic cells in glucose-free media. Blocking RIP1 signaling, by necrostatin-1 or gene silencing, decreased lactodehydrogenase (LDH) leakage and plasma membrane disintegration. Generation of mitochondrial superoxide was noted after hypoxic challenge; its reduction by antioxidants inhibited RIP signaling and cell necrosis. Supplementation of glucose diminished the RIP-dependent LDH leakage and morphological damage in hypoxic cells, whereas non-metabolizable sugar analogs did not. Hypoxic cells given glucose showed nuclear translocation of HIF1α associated with upregulation of GLUT-1 and GLUT-4 expression, as well as increase of intracellular ATP, pyruvate and lactate levels. The glucose-mediated death resistance was ablated by iodoacetate (an inhibitor to glyceraldehyde-3-phosphate dehydrogenase), but not by UK5099 (an inhibitor to mitochondrial pyruvate carrier), suggesting that glycolytic pathway was involved in anti-necrotic mechanism. Lastly, replacing glucose with cell-permeable pyruvate derivative also led to decrease of Hx-induced necroptosis by suppression of mitochondrial superoxide in an energy-independent manner. In conclusion, glycolytic metabolism confers resistance to RIP-dependent necroptosis in hypoxic cancer cells partly through pyruvate scavenging of mitochondrial free radicals.
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影响因子：
56.9
作者：
Jaakkola, P;Mole, DR;Ratcliffe, PJ
通讯作者：
Ratcliffe, PJ
DOI：
10.1016/j.aanat.2010.03.001
发表时间：
2010-05-20
期刊：
Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft
影响因子：
--
作者：
Airley R;Evans A;Mobasheri A;Hewitt SM
通讯作者：
Hewitt SM
影响因子：
5.5
作者：
Camara, Amadou K. S.;Aldakkak, Mohammed;Stowe, David F.
通讯作者：
Stowe, David F.
影响因子：
2.5
作者：
Kalda, A;Eriste, E;Zharkovsky, A
通讯作者：
Zharkovsky, A
影响因子：
4.4
作者：
Dufort, Fay J.;Bleiman, Blair F.;Chiles, Thomas C.
通讯作者：
Chiles, Thomas C.