2-Deoxy-D-glucose increases GFAT1 phosphorylation resulting in endoplasmic reticulum-related apoptosis via disruption of protein N-glycosylation in pancreatic cancer cells
2-Deoxy-D-glucose increases GFAT1 phosphorylation resulting in endoplasmic reticulum-related apoptosis via disruption of protein N-glycosylation in pancreatic cancer cells
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DOI：
10.1016/j.bbrc.2018.05.041
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发表时间：
2018-06-27
影响因子：
3.1
通讯作者：
Naito, Zenya
中科院分区：
文献类型：
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作者：
Ishino, Kousuke;Kudo, Mitsuhiro;Naito, Zenya
糖酵解抑制剂2-脱氧-D-葡萄糖（2DG）导致能量饥饿，影响广泛的癌细胞系中的细胞活力。为了确定2DG在胰腺癌中的作用，我们对2DG处理后的胰腺癌细胞系进行了蛋白质组学分析。80个蛋白质表现出差异表达，其中，参与磷酸己糖代谢的蛋白质被上调。谷氨酰胺上调：果糖6-磷酸氨基转移酶1（GFAT 1）属于产生尿苷二磷酸N-乙酰葡糖胺（UDP-GlcNAc）以维持糖蛋白的己糖胺生物合成途径（HBP），通过评估mRNA和蛋白质水平来验证。因此，我们评估了总N-糖蛋白的量。出乎意料的是，我们发现减少总N-糖蛋白和磷酸化的GFAT 1的AMP激活的蛋白激酶（AMPK）。这些数据可能揭示HBP功能障碍。此外，我们发现内质网（ER）应激伴随着ER应激标志物的表达增加，如葡萄糖反应蛋白78（GRP 78）和C/EBP同源蛋白（CHOP），在2DG处理的细胞。此外，在2DG存在下，二甲双胍（Met）对AMPK的加性激活协同增强了蛋白质N-糖基化的减少和细胞生长抑制。这些结果表明，2DG减少蛋白质的N-糖基化后，增加磷酸化的GFAT 1和结果在胰腺癌细胞的ER应激介导的细胞生长的抑制。(C)2018作者由爱思唯尔公司出版
The glycolytic inhibitor 2-deoxy-D-glucose (2DG) causes energy starvation, affecting cell viability in a wide range of cancer cell lines. To determine the action of 2DG in pancreatic cancer, we performed proteomic analysis of pancreatic cancer cell line after 2DG treatment. Eighty proteins showed differential expression and among these, proteins involved in phosphohexose metabolism were upregulated. Up regulation of glutamine: fructose 6-phosphate aminotransferase 1 (GFAT1), which belongs to the hexosamine biosynthesis pathway (HBP) that produces uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) to maintain glycoprotein, was validated by evaluation of mRNA and protein levels. Therefore, we assessed the amounts of total N-glycoproteins. Unexpectedly, we found a reduction of total N-glycoproteins and phosphorylation of GFAT1 by AMP-activated protein kinase (AMPK). These data may shed light on HBP dysfunction. Furthermore, we found endoplasmic reticulum (ER) stress accompanied by increased expression of ER stress markers, such as glucose response protein 78 (GRP78) and C/EBP-homologous protein (CHOP), in 2DG-treated cells. Moreover, the additive activation of AMPK by metformin (Met) synergistically enhanced the reduction of protein N-glycosylation and cell growth inhibition in the presence of 2DG. These results suggest that 2DG reduces N-glycosylation of proteins following the increase of phosphorylation of GFAT1 and results in the inhibition of cell growth mediated by ER stress in pancreatic cancer cells. (C) 2018 The Authors. Published by Elsevier Inc.