Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening
Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening
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DOI：
10.1016/j.bbrc.2015.09.106
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发表时间：
2015-11-06
影响因子：
3.1
通讯作者：
Yoshida, Minoru
中科院分区：
文献类型：
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作者：
Kobayashi, Hiroki;Nishimura, Haruna;Yoshida, Minoru
结合糖酵解抑制，与其他抗癌疗法是一个潜在的方法来治疗癌症。在这种情况下，我们试图利用针对15000个基因的shRNA文库，在癌细胞中鉴定决定对2-脱氧葡萄糖（2DG）（一种糖酵解抑制剂）敏感性的基因。筛选结果显示，在逆行转运中必不可少的COPB1和ARCN1是2DG敏感性的决定因素：沉默COPB1或ARCN1的表达使细胞对2DG毒性敏感。为了解决通过抑制copi介导的转运来增强2DG毒性的机制，我们重点研究了脂解作为糖酵解抑制的替代能量来源的作用。在脂解过程中，脂肪甘油三酯脂肪酶（ATGL）的脂滴定位需要copi介导的转运，ATGL是一种将三酰甘油作为β -氧化底物产生脂肪酸的关键酶。ATGL抑制剂atglistatin增强了2DG毒性，这与copi介导的ATGL转运到脂滴的缺陷抑制能量供应的模型一致，从而使细胞对糖酵解抑制敏感。总的来说，我们的数据表明，copi介导的转运缺陷或ATGL的药理抑制增强了癌细胞中的2DG毒性，可能是由于能量供应减少。(C) 2015爱思唯尔公司版权所有。
Combining glycolytic inhibition, with other anti-cancer therapies is a potential approach to treating cancer. In this context, we attempted to identify genes that determine sensitivity to 2-deoxyglucose (2DG), a glycolytic inhibitor, in cancer cells using pooled shRNA libraries targeting 15,000 genes. The screen revealed that COPB1 and ARCN1, which are essential in retrograde transport, as determinants of sensitivity to 2DG: silencing of COPB1 or ARCN1 expression sensitized cells to 2DG toxicity. To address the mechanism of potentiation of 2DG toxicity by inhibition of COPI-mediated transport, we focused on the role of lipolysis as an alternate source of energy upon inhibition of glycolysis. In the process of lipolysis, COPI-mediated transport is required for localization to lipid droplets of adipose triglyceride lipase (ATGL), a key enzyme that produces fatty acids from triacylglycerol as a substrate for beta-oxidation. The ATGL inhibitor atglistatin potentiated 2DG toxicity, consistent with a model in which a defect in COPI-mediated transport of ATGL to lipid droplets inhibits energy supply, thereby sensitizing cells to glycolytic inhibition. Collectively, our data demonstrated that a defect in COPI-mediated transport or pharmacological inhibition of ATGL potentiates 2DG toxicity in cancer cells, possibly due to a reduction in the energy supply. (C) 2015 Elsevier Inc. All rights reserved.