BAM15 Enhances Regorafenib Cytotoxicity in a Canine Osteosarcoma Cell Model
- 1Laboratory of Preclinical Research “In VetBio”, Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, Norwida 31, 50-375 Wroclaw, Poland
- 2Department of Clinical Science, Centre for Cancer Biomarkers, University of Bergen, Jonas Lies vei 87, 5021 Bergen, Norway
- 3Department of Oncology and Surgical Oncology for Children and Youth, Institute of Mother and Child, Kasprzaka 17a, 01-211 Warsaw, Poland
Abstract
Osteosarcoma is an aggressive primary bone malignancy affecting both humans and dogs, and canine osteosarcoma represents a valuable comparative model for investigating novel therapeutic strategies. Targeting tumour metabolism may enhance the efficacy of existing anticancer treatments. This study investigated whether the mitochondrial uncoupler BAM15 modifies the response of canine D17 osteosarcoma cells to regorafenib. Cells were treated with BAM15 (20 µM) or regorafenib (REG; IC25, 8 µM; IC50, 16 µM) alone or pretreated with BAM15 before regorafenib exposure. Treatment effects were assessed using viability assays, Annexin V/propidium iodide staining, cell-cycle analysis, Seahorse XF bioenergetic profiling, quantitative PCR, and Western blotting. BAM15 alone did not significantly affect cell metabolic activity. Combined treatment with BAM15 and regorafenib produced a significantly greater reduction in cell metabolic activity than regorafenib alone at IC50, whereas no significant difference between treatments was observed at IC25. Combined treatment at IC50 increased late apoptotic and necrotic populations and disrupted cell-cycle distribution. BAM15 reduced oxidative phosphorylation-derived ATP production without detectable glycolytic compensation but did not further suppress mitochondrial ATP production beyond regorafenib effect alone. Combined treatment also induced transcriptional changes in stress-adaptation-related genes, while protein analysis revealed reduced BAX and BAX/BCL-2 ratio. These exploratory findings provide preliminary evidence that mitochondrial uncoupling by BAM15 may modify the response to regorafenib and support further mechanistic investigation and validation in additional tumour and non-malignant canine models.
Related
- Complementary cytotoxicity of GD2-targeted photoimmunotherapy and 5-aminolevulinic acid photodynamic therapy in neuroblastoma and osteosarcoma
- Naringenin Enhances the Inhibitory Effect of Regorafenib on HepG2 Hepatocellular Carcinoma Cell Viability with Modulation of E-Cadherin and MRP1 Gene Expression
- Honokiol induces an autophagy and apoptosis imbalance by modulating the p62 and Nrf2 signaling loop in canine osteosarcoma
- A temporal coupling hypothesis for chemoresistance in canine DLBCL and osteosarcoma: stress signalling and microRNA regulation
- Immunohistochemical detection of the transcription factor osterix in canine and feline osteosarcoma
BibTeX
@article{dolnicka2026bam15,
title = {{BAM15 Enhances Regorafenib Cytotoxicity in a Canine Osteosarcoma Cell Model}},
author = {Agnieszka Dolnicka and Klaudia Marcinkowska and Bożena Obmińska-Mrukowicz and Vibeke Fosse and Anna Raciborska and Agnieszka Śmieszek},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27209121},
url = {https://www.mdpi.com/1422-0067/27/20/9121},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - BAM15 Enhances Regorafenib Cytotoxicity in a Canine Osteosarcoma Cell Model
AU - Agnieszka Dolnicka
AU - Klaudia Marcinkowska
AU - Bożena Obmińska-Mrukowicz
AU - Vibeke Fosse
AU - Anna Raciborska
AU - Agnieszka Śmieszek
PY - 2026
DA - 2026/10/14
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27209121
UR - https://www.mdpi.com/1422-0067/27/20/9121
AB - Osteosarcoma is an aggressive primary bone malignancy affecting both humans and dogs, and canine osteosarcoma represents a valuable comparative model for investigating novel therapeutic strategies. Targeting tumour metabolism may enhance the efficacy of existing anticancer treatments. This study investigated whether the mitochondrial uncoupler BAM15 modifies the response of canine D17 osteosarcoma cells to regorafenib. Cells were treated with BAM15 (20 µM) or regorafenib (REG; IC25, 8 µM; IC50, 16 µM) alone or pretreated with BAM15 before regorafenib exposure. Treatment effects were assessed using viability assays, Annexin V/propidium iodide staining, cell-cycle analysis, Seahorse XF bioenergetic profiling, quantitative PCR, and Western blotting. BAM15 alone did not significantly affect cell metabolic activity. Combined treatment with BAM15 and regorafenib produced a significantly greater reduction in cell metabolic activity than regorafenib alone at IC50, whereas no significant difference between treatments was observed at IC25. Combined treatment at IC50 increased late apoptotic and necrotic populations and disrupted cell-cycle distribution. BAM15 reduced oxidative phosphorylation-derived ATP production without detectable glycolytic compensation but did not further suppress mitochondrial ATP production beyond regorafenib effect alone. Combined treatment also induced transcriptional changes in stress-adaptation-related genes, while protein analysis revealed reduced BAX and BAX/BCL-2 ratio. These exploratory findings provide preliminary evidence that mitochondrial uncoupling by BAM15 may modify the response to regorafenib and support further mechanistic investigation and validation in additional tumour and non-malignant canine models.
SN - 1422-0067
ER -