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Updated: Jun 16, 2025
In Vivo Model for Testing Effect of Hypoxia on Tumor Metastasis
Published on: December 9, 2016
TH-302 (evofosfamide) monotherapy exerts anticancer activity in Ewing's sarcoma cells under hypoxia
Marie Kühne1,2,3, Sabine Becker1,4,3, Jürgen Sonnemann1,4,3
1Department of Pediatric and Adolescent Medicine, Jena University Hospital, Friedrich Schiller University Jena, Jena, Germany.
Background:
Tumor hypoxia is a significant factor in cancer progression, metastasis, and therapy resistance, leading to poor patient outcomes. Hypoxia-activated prodrugs (HAPs) are a class of agents that selectively target these hypoxic environments. They remain inactive under normal oxygen conditions but are activated by low oxygen levels. TH-302 (evofosfamide), a nitroimidazole mustard, is a clinically advanced HAP. This study aimed to investigate the effects of TH-302 in three Ewing's sarcoma (ES) cell lines.
Methods:
TH-302 was assessed for its effects on DNA damage, cell proliferation, cell death, mitochondrial depolarization, caspase-3/7 activation, and the emergence of sub-G1 cell populations using flow cytometry, fluorescence microscopy and quantitative real time (qRT)-PCR. These effects were compared under different oxygen concentrations.
Results:
TH-302 induced DNA damage and reduced cell number in ES cells by over 60%, preferentially under hypoxic conditions, independent of the cellular p53 status. TH-302 caused cell death in up to 40% of cells and mitochondrial depolarization in up to 60% of cells. Additionally, TH-302-induced caspase-3/7 activation and increased sub-G1 cell populations predominantly under hypoxia, with the most pronounced effects occurring at 0.2% environmental oxygen compared to 1% O2.
Conclusion:
These in vitro findings suggest that TH-302 may be efficacious in ES. This provides a rationale for further in vivo investigations into the potential of TH-302 as a treatment for ES.
