Anti-cancer agent 3-bromopyruvate reduces growth of MPNST and inhibits metabolic pathways in a representative in-vitro model

• Published in: BMC cancer Vol. 20; no. 1; pp. 896 - 11
• Main Authors: Linke, Christian, Wösle, Markus, Harder, Anja
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 18.09.2020; BioMed Central; BMC
• Subjects: 3-BrPA; Adenine; Adenosine; Analysis; Antineoplastic agents; Apoptosis; Bcl-6 protein; Cancer; Cancer treatment; Cell culture; Cell cycle; Cell Proliferation; Cyclin D1; Cytotoxicity; Dehydrogenases; Drug therapy; Enzyme Inhibitors - pharmacology; Enzyme Inhibitors - therapeutic use; Enzymes; Experiments; Fibroblasts; Glucose metabolism; Glycolysis; Growth; Humans; Lactates; Lactic acid; Metabolic Networks and Pathways; Metabolic pathways; Metabolism; Mitochondria; Mitochondrial respiration; MPNST; Mutation; NAD; NADH; NADH dehydrogenase; Nervous system; Neurofibromatosis; Neurofibrosarcoma - drug therapy; NF1; Niacinamide; p53; p53 Protein; Peripheral nerves; Physiological aspects; Proteasome activator; Pyruvates - pharmacology; Pyruvates - therapeutic use; Reactive oxygen species; Recklinghausen's disease; Respiration; Tumor proteins; Tumors; Viability; PA28; Starvation; Cell cycle; Glycolysis; Mitochondrial respiration; NF1; 3-BrPA; B8 fibroblasts; MPNST; p53
• ISSN: 1471-2407; 1471-2407
• DOI: 10.1186/s12885-020-07397-w

Anticancer compound 3-bromopyruvate (3-BrPA) suppresses cancer cell growth via targeting glycolytic and mitochondrial metabolism. The malignant peripheral nerve sheath tumor (MPNST), a very aggressive, therapy resistant, and Neurofibromatosis type 1 associated neoplasia, shows a high metabolic activity and affected patients may therefore benefit from 3-BrPA treatment. To elucidate the specific mode of action, we used a controlled cell model overexpressing proteasome activator (PA) 28, subsequently leading to p53 inactivation and oncogenic transformation and therefore reproducing an important pathway in MPNST and overall tumor pathogenesis. Viability of MPNST cell lines S462, NSF1, and T265 in response to increasing doses (0-120 μM) of 3-BrPA was analyzed by CellTiter-Blue® assay. Additionally, we investigated viability, reactive oxygen species (ROS) production (dihydroethidium assay), nicotinamide adenine dinucleotide dehydrogenase activity (NADH-TR assay) and lactate production (lactate assay) in mouse B8 fibroblasts overexpressing PA28 in response to 3-BrPA application. For all experiments normal and nutrient deficient conditions were tested. MPNST cell lines were furthermore characterized immunohistochemically for Ki67, p53, bcl2, bcl6, cyclin D1, and p21. MPNST significantly responded dose dependent to 3-BrPA application, whereby S462 cells were most responsive. Human control cells showed a reduced sensitivity. In PA28 overexpressing cancer cell model 3-BrPA application harmed mitochondrial NADH dehydrogenase activity mildly and significantly failed to inhibit lactate production. PA28 overexpression was associated with a functional glycolysis as well as a partial resistance to stress provoked by nutrient deprivation. 3-BrPA treatment was not associated with an increase of ROS. Starvation sensitized MPNST to treatment. Aggressive MPNST cells are sensitive to 3-BrPA therapy in-vitro with and without starvation. In a PA28 overexpression cancer cell model leading to p53 inactivation, thereby reflecting a key molecular feature in human NF1 associated MPNST, known functions of 3-BrPA to block mitochondrial activity and glycolysis were reproduced, however oncogenic cells displayed a partial resistance. To conclude, 3-BrPA was sufficient to reduce NF1 associated MPNST viability potentially due inhibition of glycolysis which should lead to the initiation of further studies and promises a potential benefit for NF1 patients.