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Radiation intensifies cyclophosphamide-associated ferroptosis-related changes in mouse spermatocyte-derived GC-2spd cells
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H. Jin   , Z. Xin , Q. Qian , R. Han |
| Department of Prenatal Diagnosis, Xinjiang Uygur Autonomous Region Maternal and Child Health Hospital, Urumqi 830000, Xinjiang, China , 1f10547@xjmu.edu.cn, |
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Abstract: (26 Views) |
Background: To determine whether graded X-ray irradiation intensifies cyclophosphamide (CTX)-associated cytotoxic, oxidative, iron-related, and nuclear factor erythroid 2-related factor 2 (NRF2) and glutathione peroxidase 4 (GPX4) transcriptional changes in GC-2spd mouse spermatocyte-derived cells. Materials and Methods: GC-2spd cells were evaluated in three experimental series: CTX dose-response testing (0-400 μM), irradiation alone (0, 2, 4, or 8 Gy), and irradiation on a fixed 50 μM CTX background (0, 2, 4, or 8 Gy). For the fixed-CTX series, cells were exposed to CTX for 2 h before irradiation and maintained in CTX-containing medium for 48 h. All cellular, biochemical, and transcriptional endpoints were assessed 48 h after sham irradiation or irradiation. Results: CTX and X-ray irradiation each reduced cell viability in a concentration- or dose-dependent manner. On a fixed 50 μM CTX background, increasing radiation dose further reduced viability and increased apoptosis. Irradiation also increased malondialdehyde, total iron, and Fe²⁺ levels, decreased superoxide dismutase activity, and further reduced NRF2 and GPX4 mRNA expression in CTX-exposed cells, with the clearest changes at 4 and 8 Gy. These findings indicate ferroptosis-associated biochemical and transcriptional changes, but do not establish ferroptotic cell death or a CTX-radiation interaction. Conclusion: Graded X-ray irradiation intensified CTX-associated cytotoxicity, oxidative stress, iron dysregulation, and NRF2/GPX4 transcript suppression in GC-2spd cells. Direct lipid-peroxidation assays, protein-level pathway assessment, ferroptosis-rescue experiments, validated CTX bioactivation, and a factorial interaction design are required before causal ferroptosis or radiosensitization can be concluded. |
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| Keywords: Cyclophosphamide, X-Rays, ferroptosis, spermatocytes. |
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Full-Text [PDF 1132 kb]
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Type of Study: Original Research |
Subject:
Radiation Biology
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