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Synthesis of cytochrome c oxidase 2: a p53-dependent metabolic regulator that promotes respiratory function and protects glioma and colon cancer cells from hypoxia-induced cell death

Oncogene volume 31, pages 3764–3776 (2012)Cite this article

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Abstract

P53 has an important role in the processing of starvation signals. P53-dependent molecular mediators of the Warburg effect reduce glucose consumption and promote mitochondrial function. We therefore hypothesized that the retention of wild-type p53 characteristic of primary glioblastomas limits metabolic demands induced by deregulated signal transduction in the presence of hypoxia and nutrient depletion. Here we report that short hairpin RNA-mediated gene suppression of wild-type p53 or ectopic expression of mutant temperature-sensitive dominant-negative p53V135A increased glucose consumption and lactate production, decreased oxygen consumption and enhanced hypoxia-induced cell death in p53 wild-type human glioblastoma cells. Similarly, genetic knockout of p53 in HCT116 colon carcinoma cells resulted in reduced respiration and hypersensitivity towards hypoxia-induced cell death. Further, wild-type p53 gene silencing reduced the expression of synthesis of cytochrome c oxidase 2 (SCO2), an effector necessary for respiratory chain function. An SCO2 transgene reverted the metabolic phenotype and restored resistance towards hypoxia in p53-depleted and p53 mutant glioma cells in a rotenone-sensitive manner, demonstrating that this effect was dependent on intact oxidative phosphorylation. Supplementation with methyl-pyruvate, a mitochondrial substrate, rescued p53 wild-type but not p53 mutant cells from hypoxic cell death, demonstrating a p53-mediated selective aptitude to metabolize mitochondrial substrates. Further, SCO2 gene silencing in p53 wild-type glioma cells sensitized these cells towards hypoxia. Finally, lentiviral gene suppression of SCO2 significantly enhanced tumor necrosis in a subcutaneous HCT116 xenograft tumor model, compatible with impaired energy metabolism in these cells. These findings demonstrate that glioma and colon cancer cells with p53 wild-type status can skew the Warburg effect and thereby reduce their vulnerability towards tumor hypoxia in an SCO2-dependent manner. Targeting SCO2 may therefore represent a valuable strategy to enhance sensitivity towards hypoxia and may complement strategies targeting glucose metabolism.

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Abbreviations

AMPKα:

AMP-activated protein kinase α

hygro:

hygromycin

luc:

luciferase

p53ts:

temperature-sensitive p53

phospho:

phosphorylated

PI:

propidium iodide

puro:

puromycin

ROS:

reactive oxygen species

SCO2:

synthesis of cytochrome c oxidase 2

shRNA:

short hairpin RNA

siRNA:

small interfering RNA

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Acknowledgements

We thank P Hwang for providing the SCO2 expression plasmid and B Vogelstein for the HCT116 p53+/+ and p53−/− cell lines. This work was supported by The Deutsche Forschungsgemeinschaft (GRK 1302/1). The Dr Senckenberg Institute of Neurooncology is supported by the Hertie foundation and the Dr Senckenberg foundation. JPS is ‘Hertie Professor for Neurooncology’.

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Authors and Affiliations

  1. Dr Senckenberg Institute of Neurooncology, Goethe University Frankfurt, Frankfurt, Germany

    C Wanka, D P Brucker, O Bähr, M Ronellenfitsch, J P Steinbach & J Rieger

  2. Department of Neurology, University Hospital Tübingen, Tübingen, Germany

    C Wanka, D P Brucker & J P Steinbach

  3. Department of Neurology, University Hospital Zürich, Zürich, Switzerland

    M Weller

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  1. C Wanka
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Wanka, C., Brucker, D., Bähr, O. et al. Synthesis of cytochrome c oxidase 2: a p53-dependent metabolic regulator that promotes respiratory function and protects glioma and colon cancer cells from hypoxia-induced cell death. Oncogene 31, 3764–3776 (2012). https://doi.org/10.1038/onc.2011.530

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