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The plasma membrane redox enzyme NQO1 sustains cellular energetics and protects human neuroblastoma cells against metabolic and proteotoxic stress

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Abstract

The plasma membrane redox system (PMRS) of nicotinamide adenine dinucleotide (NADH)-related enzymes plays a key role in the maintenance of cellular energetics. During the aging process, neural cells are particularly sensitive to impaired energy metabolism and oxidative damage, but the involvement of the PMRS in these processes is unknown. Here, we used human neuroblastoma cells with either elevated or reduced levels of the PMRS enzyme NADH-quinone oxidoreductase 1 (NQO1) to investigate how the PMRS regulates neuronal stress responses. Cells with elevated NQO1 levels were more resistant to death induced by 2-deoxyglucose, potassium cyanide (energetic stress), and lactacystin (proteotoxic stress), but were not protected from being killed by H2O2 and serum withdrawal. The NAD+(an oxidized form of NADH)/NADH ratio was maintained at a significantly higher level in cells overexpressing NQO1, consistent with enhanced levels of NQO1 activity. Levels of the neuroprotective transcription factors nuclear factor kappa-light-chain-enhancer of activated B cells and nuclear factor (erythroid-derived 2)-like 2, and the protein chaperone HSP70 were elevated in cells overexpressing NQO1. Cells in which NQO1 levels were decreased by RNA interference exhibited increased vulnerability to death induced by 2-deoxyglucose and lactacystin. Thus, a higher NAD+/NADH ratio and activation of adaptive stress response pathways are enhanced by the PMRS in neuroblastoma cells, enabling them to maintain redox homeostasis under conditions of energetic and proteotoxic stress. These findings have implications for the development of therapeutic interventions for neural tumors and neurodegenerative conditions.

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Acknowledgments

We thank Alan Sartorelli (Yale University School of Medicine, USA) and David Ross (University of Colorado at Denver, USA) for providing pBE8 and antibodies against NQO1, respectively. This study was supported, in whole and in part, by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (20100003064), South Korea.

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Authors and Affiliations

  1. Division of Life and Pharmaceutical Sciences, Department of Life Science, Ewha Womans University, Seoul, 120–750, South Korea

    Dong-Hoon Hyun & Jiyeong Kim

  2. Cell Therapy Center, Hanyang Univeristy Seoul Hospital, Seoul, 133-792, South Korea

    Chanil Moon

  3. Division of Life Sciences, Kangwon National University, Chuncheon, 200–701, South Korea

    Chang-Jin Lim

  4. Laboratory of Experimental Gerontology, National Institute on Aging Intramural Research Program, National Institutes of Health, Baltimore, MD, 21224, USA

    Rafael de Cabo

  5. Laboratory of Neurosciences, National Institute on Aging Intramural Research Program, National Institutes of Health, Baltimore, MD, 21224, USA

    Mark P. Mattson

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  1. Dong-Hoon Hyun
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  2. Jiyeong Kim
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Correspondence to Dong-Hoon Hyun.

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Hyun, DH., Kim, J., Moon, C. et al. The plasma membrane redox enzyme NQO1 sustains cellular energetics and protects human neuroblastoma cells against metabolic and proteotoxic stress. AGE 34, 359–370 (2012). https://doi.org/10.1007/s11357-011-9245-1

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