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01-01-2017 | Original Article

Normoxic accumulation of HIF1α is associated with glutaminolysis

Authors: Matthias Kappler, Ulrike Pabst, Swetlana Rot, Helge Taubert, Henri Wichmann, Johannes Schubert, Matthias Bache, Claus Weinholdt, Uta-Dorothee Immel, Ivo Grosse, Dirk Vordermark, Alexander W. Eckert

Published in: Clinical Oral Investigations | Issue 1/2017

Abstract

Objectives

The stabilization of the transcription factor and prognostic tumor marker hypoxia-inducible factor 1α (HIF1α) is considered to be crucial for cellular metabolic adaptations to hypoxia. However, HIF1α has also been shown to accumulate under normoxic conditions, although this phenomenon is poorly understood.

Methods

We investigated the conditions for normoxic HIF1α stabilization in different tumor cell lines (e.g., two mammary carcinoma cell lines and three oral squamous cell carcinoma cell lines) via Western blot analysis or immunohistochemical staining. The transcriptional activity of HIF1 was demonstrated by analyzing the messenger RNA (mRNA) expression of the HIF1 target carbonic anhydrase 9 (CA9) via PCR.

Results

Our data demonstrate that the combined incubation of tumor cells with glutamine and growth factors (e.g., EGF, insulin, and serum) mediates the normoxic accumulation of HIF1α in vitro. Consequently, the inhibition of glutaminolysis by a glutaminase inhibitor blocked the normoxic accumulation of HIF1α. Additionally, the normoxic HIF1α protein displayed nuclear translocation and transcriptional activity, which was confirmed by the induction of CA9 mRNA expression. Furthermore, the normoxic accumulation of HIF1α was associated with impaired proliferation of tumor cells. Finally, ammonia, the toxic waste product of glutaminolysis, induced a normoxic accumulation of HIF1α to the same extent as glutamine.

Conclusion

Our study suggests that HIF1α is involved in the regulation of glutamine metabolism and the cellular levels of the toxic metabolic waste product ammonia under normoxia. Hence, our results, together with data presented in the literature, support the hypothesis that HIF1α and its target genes play a crucial role in metabolic pathways, such as glutaminolysis and glycolysis, under both hypoxic and normoxic conditions.

Clinical relevance

Therefore, the inhibition of HIF1α (and/or HIF1α target genes) could emerge as a promising therapeutic approach that would result in the accumulation of toxic metabolic waste products in tumor cells as well as the reduction of their nutrition and energy supply.

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Title: Normoxic accumulation of HIF1α is associated with glutaminolysis
Authors: Matthias Kappler
Ulrike Pabst
Swetlana Rot
Helge Taubert
Henri Wichmann
Johannes Schubert
Matthias Bache
Claus Weinholdt
Uta-Dorothee Immel
Ivo Grosse
Dirk Vordermark
Alexander W. Eckert
Publication date: 01-01-2017
Publisher: Springer Berlin Heidelberg
Published in: Clinical Oral Investigations / Issue 1/2017
Print ISSN: 1432-6981
Electronic ISSN: 1436-3771
DOI: https://doi.org/10.1007/s00784-016-1780-9

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Normoxic accumulation of HIF1α is associated with glutaminolysis

Abstract

Objectives

Methods

Results

Conclusion

Clinical relevance

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Objectives

Methods

Results

Conclusion

Clinical relevance

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