Effects of the potassium-sparing diuretic amiloride on chemotherapy response in canine osteosarcoma cells.
Amiloride
/ administration & dosage
Animals
Antineoplastic Agents
/ administration & dosage
Apoptosis
/ drug effects
Bone Neoplasms
/ drug therapy
Carboplatin
/ administration & dosage
Cell Line, Tumor
Cell Proliferation
/ drug effects
Dog Diseases
/ drug therapy
Dogs
Doxorubicin
/ administration & dosage
Drug Therapy, Combination
/ veterinary
Osteosarcoma
/ drug therapy
Proton Pump Inhibitors
/ administration & dosage
acidification
chemoresistance
dog
doxorubicin
proton pumps
sensitization
Journal
Journal of veterinary internal medicine
ISSN: 1939-1676
Titre abrégé: J Vet Intern Med
Pays: United States
ID NLM: 8708660
Informations de publication
Date de publication:
Mar 2019
Mar 2019
Historique:
received:
08
05
2018
accepted:
14
11
2018
pubmed:
18
12
2018
medline:
11
5
2019
entrez:
18
12
2018
Statut:
ppublish
Résumé
Osteosarcoma (OSA) is a common bone tumor of mesenchymal origin in dogs. Chemotherapy delays metastasis, yet most dogs die of this disease within 1 year of diagnosis. The high metabolic demand of cancer cells promotes proton pump upregulation, leading to acidification of the tumor microenvironment and chemoresistance. The potassium-sparing diuretic amiloride is among a class of proton pump inhibitors prescribed for refractory heart failure treatment in dogs. We hypothesized that amiloride treatment improves chemotherapy response by reducing acidification in canine OSA cells. Our objective was to assess the in vitro effects of amiloride on cell viability, apoptosis, and metabolism. In vitro study. Assessments of cell viability and apoptosis were performed after single agent or combination treatment, along with calculations of pharmacological synergism using the combination index. Protein signaling during apoptosis was evaluated by Western blotting. Metabolic profiling was performed using a Seahorse bioanalyzer. Amiloride strongly synergized with doxorubicin in combination treatment and exhibited additive or antagonistic effects with carboplatin in canine OSA cells. Combination treatment with doxorubicin significantly upregulated p53-mitochondrial signaling to activate apoptosis and downregulate Akt phosphorylation. Amiloride-treated cells further exhibited metabolic switching with reductions in glycolytic capacity and maximal respiration. Amiloride synergized with doxorubicin to potentiate apoptosis in canine OSA cells. These results justify further investigation into repurposing of amiloride as an oncology drug for the treatment of OSA in dogs.
Sections du résumé
BACKGROUND
BACKGROUND
Osteosarcoma (OSA) is a common bone tumor of mesenchymal origin in dogs. Chemotherapy delays metastasis, yet most dogs die of this disease within 1 year of diagnosis. The high metabolic demand of cancer cells promotes proton pump upregulation, leading to acidification of the tumor microenvironment and chemoresistance. The potassium-sparing diuretic amiloride is among a class of proton pump inhibitors prescribed for refractory heart failure treatment in dogs.
OBJECTIVE
OBJECTIVE
We hypothesized that amiloride treatment improves chemotherapy response by reducing acidification in canine OSA cells. Our objective was to assess the in vitro effects of amiloride on cell viability, apoptosis, and metabolism.
METHODS
METHODS
In vitro study. Assessments of cell viability and apoptosis were performed after single agent or combination treatment, along with calculations of pharmacological synergism using the combination index. Protein signaling during apoptosis was evaluated by Western blotting. Metabolic profiling was performed using a Seahorse bioanalyzer.
RESULTS
RESULTS
Amiloride strongly synergized with doxorubicin in combination treatment and exhibited additive or antagonistic effects with carboplatin in canine OSA cells. Combination treatment with doxorubicin significantly upregulated p53-mitochondrial signaling to activate apoptosis and downregulate Akt phosphorylation. Amiloride-treated cells further exhibited metabolic switching with reductions in glycolytic capacity and maximal respiration.
CONCLUSION AND CLINICAL IMPORTANCE
CONCLUSIONS
Amiloride synergized with doxorubicin to potentiate apoptosis in canine OSA cells. These results justify further investigation into repurposing of amiloride as an oncology drug for the treatment of OSA in dogs.
Identifiants
pubmed: 30556178
doi: 10.1111/jvim.15382
pmc: PMC6430882
doi:
Substances chimiques
Antineoplastic Agents
0
Proton Pump Inhibitors
0
Amiloride
7DZO8EB0Z3
Doxorubicin
80168379AG
Carboplatin
BG3F62OND5
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
800-811Subventions
Organisme : Memorial donation through University of Guelph Alumni Affairs and Development
Organisme : OVC Pet Trust
ID : 053524
Informations de copyright
© 2018 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.
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