1. Identification of the determinants of 2-deoxyglucose sensitivity in cancer cells by shRNA library screening.
- Author
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Kobayashi H, Nishimura H, Matsumoto K, and Yoshida M
- Subjects
- Antineoplastic Agents pharmacology, Biological Transport, Active, Cell Line, Tumor, Coatomer Protein antagonists & inhibitors, Coatomer Protein genetics, Coatomer Protein metabolism, Deoxyglucose administration & dosage, Drug Resistance, Neoplasm genetics, Drug Synergism, Enzyme Inhibitors administration & dosage, Enzyme Inhibitors pharmacology, Gene Knockdown Techniques, Gene Library, Glycolysis drug effects, Glycolysis genetics, HCT116 Cells, HeLa Cells, Humans, Lipase antagonists & inhibitors, Lipase metabolism, Lipid Metabolism drug effects, Lipid Metabolism genetics, Neoplasms metabolism, Phenylurea Compounds administration & dosage, Phenylurea Compounds pharmacology, Deoxyglucose pharmacology, Neoplasms drug therapy, Neoplasms genetics, RNA, Small Interfering genetics
- Abstract
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 β-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., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
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