Abstract
Background
Inhibition of glucose metabolism has recently become an attractive target for cancer treatment. Accordingly, since 2-deoxyglucose (2-DG) competes effectively with glucose, it has come under increasing scrutiny as a therapeutic agent. The initial response of tumor cells to 2-DG is growth inhibition, which is thought to conserve energy and consequently protect cells from its ATP-lowering effects as a glycolytic inhibitor. However, since 2-DG also mimics mannose and thereby interferes with N-linked glycosylation, the question is raised of how this sugar analog inhibits tumor cell growth and whether the mechanism by which it protects cells can be manipulated to convert 2-DG-induced growth inhibition to cell death.
Methods
Cell growth and death were measured via counting viable and dead cells based on trypan blue exclusion. Markers of ATP reduction and the unfolded protein response (UPR) were detected by Western blot. Protein functions were manipulated through chemical compounds, siRNA and the use of gene-specific wild-type and knock-out mouse embryonic fibroblasts (MEFs).
Results
At 2-DG concentrations that can be achieved in human plasma without causing significant side effects, we find (a) It induces growth inhibition predominantly by interference with glycosylation, which leads to accumulation of unfolded proteins in the endoplasmic reticulum activating the UPR; (b) Inhibition of PERK (but not ATF6 or IRE1), a major component of the UPR, leads to conversion of 2-DG-induced growth inhibition to cell death and (c) secondarily to PERK, inhibition of GCN2, a kinase that is activated in response to low intracellular glutamine, increases 2-DG’s cytotoxic effects in PERK −/− MEFs.
Conclusions
Overall, these findings present a novel anticancer strategy that can be translated into therapeutic gain as they uncover the metabolic target PERK, and to a lesser degree GCN2, that when inhibited convert 2-DG’s static effect to a toxic one in tumor cells growing under normoxia.
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Abbreviations
- 2-DG:
-
2-Deoxyglucose
- UPR:
-
Unfolded protein response
- MEF:
-
Mouse embryonic fibroblast
- AMPK:
-
AMP-activated protein kinase
- ER:
-
Endoplasmic reticulum
- PPS:
-
Pentose phosphate shunt
- ROS:
-
Reactive oxygen species
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Acknowledgments
We thank Dr. David Ron (University of Cambridge, Cambridge, UK) for the PERK and IRE1 wild-type and knock-out MEF cell pairs. We also want to thank Dr. Glen Barber (University of Miami, Miami, FL) for providing us with the eIF2α wild-type and serine51 mutant MEFs. This study is supported by the National Cancer Institute grant CA37109 and Pap Corps award to TJL.
Conflict of interest
RK and JMA are employees of GlaxoSmithKline. HL, MK, YC, HX and TJL declare that they have no competing interests.
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Liu, H., Kurtoglu, M., Cao, Y. et al. Conversion of 2-deoxyglucose-induced growth inhibition to cell death in normoxic tumor cells. Cancer Chemother Pharmacol 72, 251–262 (2013). https://doi.org/10.1007/s00280-013-2193-y
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DOI: https://doi.org/10.1007/s00280-013-2193-y