title: Synergistic Anticancer Action of Lysosomal Membrane Permeabilization and Glycolysis Inhibition.
abstract: We investigated the in vitro and in vivo anticancer effect of combining lysosomal membrane permeabilization (LMP)-inducing agent N-dodecylimidazole (NDI) with glycolytic inhibitor 2-deoxy-d-glucose (2DG). NDI-triggered LMP and 2DG-mediated glycolysis block synergized in inducing rapid ATP depletion, mitochondrial damage, and reactive oxygen species production, eventually leading to necrotic death of U251 glioma cells but not primary astrocytes. NDI/2DG-induced death of glioma cells was partly prevented by lysosomal cathepsin inhibitor E64 and antioxidant α-tocopherol, suggesting the involvement of LMP and oxidative stress in the observed cytotoxicity. LMP-inducing agent chloroquine also displayed a synergistic anticancer effect with 2DG, whereas glucose deprivation or glycolytic inhibitors iodoacetate and sodium fluoride synergistically cooperated with NDI, thus further indicating that the anticancer effect of NDI/2DG combination was indeed due to LMP and glycolysis block. The two agents synergistically induced ATP depletion, mitochondrial depolarization, oxidative stress, and necrotic death also in B16 mouse melanoma cells. Moreover, the combined oral administration of NDI and 2DG reduced in vivo melanoma growth in C57BL/6 mice by inducing necrotic death of tumor cells, without causing liver, spleen, or kidney toxicity. Based on these results, we propose that NDI-triggered LMP causes initial mitochondrial damage that is further increased by 2DG due to the lack of glycolytic ATP required to maintain mitochondrial health. This leads to a positive feedback cycle of mitochondrial dysfunction, ATP loss, and reactive oxygen species production, culminating in necrotic cell death. Therefore, the combination of LMP-inducing agents and glycolysis inhibitors seems worthy of further exploration as an anticancer strategy.
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Milica
authors: M
authors: Kosic
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Katarina
authors: K
authors: Arsikin-Csordas
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Verica
authors: V
authors: Paunovic
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Raymond A
authors: RA
authors: Firestone
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Biljana
authors: B
authors: Ristic
authors: the Institute of Histology and Embryology, School of Medicine, University of Belgrade, Visegradska 26, 11000 Belgrade, Serbia.
authors: Aleksandar
authors: A
authors: Mircic
authors: the Institute of Biomedical Research, Galenika a.d., Pasterova 2, 11000 Belgrade, Serbia, and.
authors: Sasa
authors: S
authors: Petricevic
authors: the Institute of Histology and Embryology, School of Medicine, University of Belgrade, Visegradska 26, 11000 Belgrade, Serbia.
authors: Mihajlo
authors: M
authors: Bosnjak
authors: the Institute for Biological Research, University of Belgrade, Despot Stefan Blvd. 142, 11000 Belgrade, Serbia.
authors: Nevena
authors: N
authors: Zogovic
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia.
authors: Milos
authors: M
authors: Mandic
authors: the Institute of Histology and Embryology, School of Medicine, University of Belgrade, Visegradska 26, 11000 Belgrade, Serbia.
authors: Vladimir
authors: V
authors: Bumbasirevic
authors: From the Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr. Subotica 1, 11000 Belgrade, Serbia, vtrajkovic@med.bg.ac.rs.
authors: Vladimir
authors: V
authors: Trajkovic
authors: the Institute for Biological Research, University of Belgrade, Despot Stefan Blvd. 142, 11000 Belgrade, Serbia buajk@yahoo.com.
authors: Ljubica
authors: L
authors: Harhaji-Trajkovic
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D003847:Deoxyglucose / Q000494:pharmacology*; D004357:Drug Synergism; D005910:Glioma / Q000188:drug therapy / Q000378:metabolism* / Q000503:physiopathology; D006019:Glycolysis / Q000187:drug effects*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D008247:Lysosomes / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects
chemical_list: D007093:Imidazoles; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; C002822:1-dodecylimidazole
keywords: 2-deoxy-d-glucose; ATP; N-dodecylimidazole; cancer therapy; glycolysis; lysosome; mitochondrial membrane potential; necrosis (necrotic death); oxidative stress