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Amifostine induces anaerobic metabolism and hypoxia-inducible factor 1α

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Abstract

Purpose

The cytoprotective mechanism of amifostine (WR-2721) implies free radical scavenging and DNA repair activities. We investigated additional cytoprotective pathways involving intracellular hypoxia and the activation of the hypoxia-inducible factor (HIF) pathway, a key transcription factor regulating glycolysis, angiogenesis and apoptosis, which is also linked with radioresistance.

Materials and methods

The glucose and oxygen levels in the peripheral blood of patients receiving 1000 mg amifostine were determined at various time-points in order to investigate the metabolic changes induced by amifostine. MDA468 breast tumor cell lines were incubated with a high amifostine concentration (10 mM) to overcome the natural resistance of cancer cells to influx of the non-hydrolyzed WR-2721, and the HIF1α protein levels were determined by Western blot analysis. In vivo experiments with Wistar rats were performed in order to assess immunohistochemically changes in the intracellular accumulation of HIF1α induced by amifostine (200 mg/kg).

Results

By 30 min following amifostine administration, the hemoglobin oxygen saturation and pO2 levels had increased in the peripheral blood while glucose levels had reduced, providing evidence that normal tissue metabolism switches to glycolytic pathways. Incubation of cell lines with amifostine resulted in HIF1α induction. In Wistar rats administration of amifostine resulted in increased HIF1α accumulation in normal tissues.

Conclusions

Since it is doubtful whether dephosphorylation of amifostine to the active metabolite WR-1065 occurs within tumoral tissues (an acidic environment that lacks vascular alkaline phosphatase activity), intracellular hypoxia and upregulation of HIF1α represents an additional, normal tissue-specific, amifostine cytoprotective pathway.

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Authors and Affiliations

  1. Departments of Radiotherapy/Oncology, Pathology and Surgery, Democritus University of Thrace, 68100, Alexandroupolis, Greece

    Michael I. Koukourakis, Alexandra Giatromanolaki, Costantinos Simopoulos, Alexandros Polychronidis & Efthimios Sivridis

  2. Cancer Research UK, Molecular Oncology Laboratories, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, OX3 9DS, UK

    Wen Chong & Adrian L. Harris

Authors
  1. Michael I. Koukourakis
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  2. Alexandra Giatromanolaki
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  3. Wen Chong
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  4. Costantinos Simopoulos
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  5. Alexandros Polychronidis
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  6. Efthimios Sivridis
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  7. Adrian L. Harris
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Correspondence to Michael I. Koukourakis.

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This work is presented on behalf of the Tumour and Angiogenesis Research Group.

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Koukourakis, M.I., Giatromanolaki, A., Chong, W. et al. Amifostine induces anaerobic metabolism and hypoxia-inducible factor 1α. Cancer Chemother Pharmacol 53, 8–14 (2004). https://doi.org/10.1007/s00280-003-0691-z

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  • DOI: https://doi.org/10.1007/s00280-003-0691-z

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