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Aglycemia keeps mitochondrial oxidative phosphorylation under hypoxic conditions in HepG2 cells

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Abstract

Cancer cell bioenergetics, maintaining mixed aerobic glycolysis (Warburg phenotype) and oxidative phosphorylation (OXPHOS), is not fully elucidated. Hypoxia-dependent OXPHOS suppression determines aerobic glycolysis. To elucidate further details, we studied hypoxic adaptation (up to 72 h at 5 % oxygen) of hepatocellular carcinoma HepG2 cells. The key regulatory component, hypoxia-inducible factor (HIF)-1α (HIF-1α) was stabilized at 5 h in 5 % oxygen for all three studied regimens, i.e. in glycolytic cells at 5 mM or 25 mM glucose, or in aglycemic (Oxphos) cells when glucose was replaced by galactose. However, the conventional HIF-mediated suppression of respiration was prevented at aglycemia, which correlated with a high proportion of unphosphorylated pyruvate dehydrogenase (PDH) at 5 % oxygen. Such a modified HIF response in Oxphos cells, termed as a non-canonical one, contrasted to conventional respiration suppression down to 45 % or 43 %, observed in hypoxia-adapted glycolytic cells at 5 mM or 25 mM glucose, respectively. These hypoxic glycolytic cells had normally highly phosphorylated PDH and most likely utilized pyruvate by aminotransferase reaction of glutaminolysis to feed at least suppressed respiration. Also, glycolytic cells were rather resistant towards the staurosporine-induced apoptosis, whereas aglycemic (Oxphos) HepG2 cells exhibited much higher susceptibility. We conclude that aglycemia modulates the hypoxic HIF signaling toward a non-canonical response that is unable to carry out complete PDH phosphorylation, allowing a high pyruvate input for OXPHOS from the elevated glycolysis, which together with ongoing glutaminolysis maintain a virtually unchanged respiration. Similar OXPHOS revival may explain distinct tumor sensitivity to chemotherapy and other pharmacological interventions.

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Acknowledgments

This project was supported by a grants from the Grant Agency of the Czech Republic No. 302/10/0346 and No. 13-02033 to P.J., Czech Ministry of Education grant No. LH11055 to L.P.H., and the institutional support projects AV0Z50110509 and RVO67985823. We gratefully acknowledge the help of Dr. Katarína Smolková with cytochrome c oxidase kinetics and citrate synthase activity measurements and the excellent technical assistance of Lenka Josková with cell cultures and Jana Vaicová with immunoblotting.

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  1. Department of Membrane Transport Biophysics, No. 75, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague 4, Czech Republic

    Lydie Plecitá-Hlavatá, Jan Ježek & Petr Ježek

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  1. Lydie Plecitá-Hlavatá
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Plecitá-Hlavatá, L., Ježek, J. & Ježek, P. Aglycemia keeps mitochondrial oxidative phosphorylation under hypoxic conditions in HepG2 cells. J Bioenerg Biomembr 47, 467–476 (2015). https://doi.org/10.1007/s10863-015-9628-6

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