Targeting Glycolysis with 2-Deoxy-D-Glucose and Lysosomal Integrity with L-Leucyl-L-Leucine Methyl Ester as Antimelanoma Strategy

Background/Objectives: Melanoma cells enhance glycolysis and expand lysosomes to support energy metabolism, proliferation, and metastasis. However, lysosomal membrane permeabilization (LMP) causes cathepsin leakage into cytosol triggering cytotoxicity. This study investigated the antimelanoma effect of 2-deoxy-D-glucose (2DG), an inhibitor of glycolytic enzyme hexokinase-2, in combination with cathepsin C-dependent LMP inducer L-leucyl-L-leucine methyl ester (LLOMe) and cathepsin C-independent LMP-inducers mefloquine and siramesine. Methods: The viability of A375 and B16 melanoma cells and primary fibroblasts was measured by crystal violet. Apoptosis, necrosis, and LMP were assessed by flow cytometry; caspase activation, mitochondrial depolarization, superoxide production, and energy metabolism were analyzed by fluorimetry, and expression of cathepsins and hexokinase-2 was evaluated by immunoblot. Appropriate inhibitors, antioxidant, and energy boosters were used to confirm cell death type and mechanism. Results: LLOMe triggered LMP, mitochondrial depolarization, and mitochondrial superoxide production, while suppressing oxidative phosphorylation. 2DG suppressed glycolysis and, together with LLOMe, synergized in ATP depletion, caspase activation, and mixed apoptosis and necrosis in A375 cells. Inhibitors of lysosomal acidification, cysteine cathepsins, and caspases, as well as antioxidant and energy boosters, reduced 2DG+LLOMe-induced toxicity. Cathepsins B, C, and D were lower, while hexokinase-2 was higher in A375 cells than fibroblasts. Accordingly, 2DG exhibited lower while LLOMe exhibited higher toxicity against fibroblasts than A375 and B16 cells. However, mefloquine and siramesine induced stronger LMP in A375 cells than in fibroblasts and showed melanoma-selective toxicity when combined with 2DG. Conclusions: 2DG-mediated glycolysis inhibition in combination with lysosomal destabilization induced by mefloquine and siramesine, but not with non-selectively toxic LLOMe, may be promising antimelanoma strategy.

• Publication year

  2025

• Venue

  Pharmaceutics

• Publication date

  2025-10-01

• Fields of study

  Medicine

• Identifiers
  DOI 10.3390/pharmaceutics17101312PMID 41155949PMCID 12566822
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Pharmacological CDK4/6 inhibition promotes vulnerability to lysosomotropic agents in breast cancer

  2025cited by this paper
• Long non-coding RNA CYTOR promotes the progression of melanoma via the miR-485-5p/GPI axis

  2025cited by this paper
• Structural Elucidation and Antiviral Activity of Covalent Cathepsin L Inhibitors

  2024cited by this paper
• Iron retardation in lysosomes protects senescent cells from ferroptosis

  2024cited by this paper
• ROS-mediated lysosomal membrane permeabilization and autophagy inhibition regulate bleomycin-induced cellular senescence

  2024cited by this paper
• Lysosome-Disrupting Agents in Combination with Venetoclax Increase Apoptotic Response in Primary Chronic Lymphocytic Leukemia (CLL) Cells Mediated by Lysosomal Cathepsin D Release and Inhibition of Autophagy

  2024cited by this paper
• Correction for: Iron retardation in lysosomes protects senescent cells from ferroptosis

  2024cited by this paper
• Bongkrekic acid inhibits 2-deoxygulcose-induced apoptosis, leading to enhanced cytotoxicity and necrotic cell death

  2024cited by this paper
• Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries

  2024influential reference
• Identity, structure, and function of the mitochondrial permeability transition pore: controversies, consensus, recent advances, and future directions

  2023cited by this paper
• Linear ubiquitination at damaged lysosomes induces local NF-κB activation and controls cell survival

  2023cited by this paper
• Cationic amphiphilic drugs induce accumulation of cytolytic lysoglycerophospholipids in the lysosomes of cancer cells and block their recycling into common membrane glycerophospholipids

  2023cited by this paper
• The Combined Metabolically Oriented Effect of Fucoidan from the Brown Alga Saccharina cichorioides and Its Carboxymethylated Derivative with 2-Deoxy-D-Glucose on Human Melanoma Cells

  2023cited by this paper
• Succinate oxidation rescues mitochondrial ATP synthesis at high temperature in Drosophila melanogaster

  2023cited by this paper
• Lysosomal function and intracellular position determine the malignant phenotype in malignant melanoma.

  2023cited by this paper
• Alpha-Enolase (ENO1) Correlates with Invasiveness of Cutaneous Melanoma—An In Vitro and a Clinical Study

  2022cited by this paper
• Differential Effects of 2-Deoxyglucose and Glucose Deprivation on 4-Hydroxynonenal Dependent Mitochondrial Dysfunction in Primary Neurons

  2022cited by this paper
• Engaging the Lysosome and Lysosome-Dependent Cell Death in Cancer

  2022cited by this paper
• In vitro reconstitution of calcium-dependent recruitment of the human ESCRT machinery in lysosomal membrane repair

  2022cited by this paper
• The Lysosome in Malignant Melanoma: Biology, Function and Therapeutic Applications

  2022influential reference
• α-Enolase inhibits apoptosis and promotes cell invasion and proliferation of skin cutaneous melanoma

  2022cited by this paper
• Histological Skin Remodeling Following Autologous Fibroblast Application.

  2022cited by this paper
• Dieckol Inhibits Autophagic Flux and Induces Apoptotic Cell Death in A375 Human Melanoma Cells via Lysosomal Dysfunction and Mitochondrial Membrane Impairment

  2022cited by this paper
• Lysosomal damage drives mitochondrial proteome remodelling and reprograms macrophage immunometabolism

  2022influential reference
• Imiquimod-induced ROS production causes lysosomal membrane permeabilization and activates caspase-8-mediated apoptosis in skin cancer cells.

  2022cited by this paper
• Independent Drug Action in Combination Therapy: Implications for Precision Oncology

  2022cited by this paper
• Cytotoxic activity of high dose ascorbic acid is enhanced by 2-deoxy-d-glucose in glycolytic melanoma cells.

  2021cited by this paper
• Autophagy inhibitors increase the susceptibility of KRAS-mutant human colorectal cancer cells to a combined treatment of 2-deoxy-D-glucose and lovastatin

  2021cited by this paper
• 3-Methyladenine prevents energy stress-induced necrotic death of melanoma cells through autophagy-independent mechanisms.

  2021cited by this paper
• Mefloquine as a dual inhibitor of glioblastoma angiogenesis and glioblastoma via disrupting lysosomal function.

  2021cited by this paper
• The glycolysis inhibitor 2-deoxyglucose ameliorates adjuvant-induced arthritis by regulating macrophage polarization in an AMPK-dependent manner.

  2021cited by this paper
• Dual Covalent Inhibition of PKM and IMPDH Targets Metabolism in Cutaneous Metastatic Melanoma

  2021cited by this paper
• Structural and Biochemical Analysis of the Dual Inhibition of MG-132 against SARS-CoV-2 Main Protease (Mpro/3CLpro) and Human Cathepsin-L

  2021cited by this paper
• Dependence of glucose transport in neuronal model systems on autophagy and GAPDH (glyceraldehyde-3 phosphate dehydrogenase) activity.

  2021cited by this paper
• Triamterene induces autophagic degradation of lysosome by exacerbating lysosomal integrity

  2021cited by this paper
• Restoration of lysosomal function after damage is accompanied by recycling of lysosomal membrane proteins

  2020influential reference
• Trifluoperazine prolongs the survival of experimental brain metastases by STAT3-dependent lysosomal membrane permeabilization.

  2020cited by this paper
• Intracellular cathepsin C levels determine sensitivity of cells to leucyl‐leucine methyl ester‐triggered apoptosis

  2020influential reference
• Pre-clinical modeling of cutaneous melanoma

  2020cited by this paper
• Cationic amphiphilic drugs induce elevation in lysoglycerophospholipid levels and cell death in leukemia cells

  2020cited by this paper
• Nanoparticle-Mediated Delivery of 2-Deoxy-D-Glucose Induces Antitumor Immunity and Cytotoxicity in Liver Tumors in Mice

  2020cited by this paper
• Dual targeting of tumor cell energy metabolism and lysosomes as an anticancer strategy.

  2020influential reference
• Cathepsin E expression and activity: Role in the detection and treatment of pancreatic cancer.

  2019cited by this paper
• Cathepsin L Regulates Metabolic Networks Controlling Rapid Cell Growth and Proliferation

  2019cited by this paper
• 2-Deoxy-d-Glucose and Its Analogs: From Diagnostic to Therapeutic Agents

  2019cited by this paper
• Disruption of Endolysosomal RAB5/7 Efficiently Eliminates Colorectal Cancer Stem Cells.

  2019cited by this paper
• Prognostic implications of markers of the metabolic phenotype in human cutaneous melanoma

  2019cited by this paper
• Galectin-3 Coordinates a Cellular System for Lysosomal Repair and Removal.

  2019cited by this paper
• 2‐Deoxy‐D‐glucose has distinct and cell line‐specific effects on the survival of different cancer cells upon antitumor drug treatment

  2018cited by this paper
• Melanoma tumors exhibit a variable but distinct metabolic signature

  2018cited by this paper
• Increased expression and altered subcellular distribution of cathepsin B in microglia induce cognitive impairment through oxidative stress and inflammatory response in mice

  2018cited by this paper
• Mechanisms of resistance to BRAF and MEK inhibitors and clinical update of US Food and Drug Administration-approved targeted therapy in advanced melanoma

  2018cited by this paper
• Bosutinib, an SRC inhibitor, induces caspase‐independent cell death associated with permeabilization of lysosomal membranes in melanoma cells

  2018cited by this paper
• Metastasis Suppressor NME1 Directly Activates Transcription of the ALDOC Gene in Melanoma Cells

  2018cited by this paper
• Targeting energy metabolism of cancer cells: Combined administration of NCL-240 and 2-DG.

  2017cited by this paper
• Mitochondrial membrane potential.

  2017cited by this paper
• Therapeutic effects of antibiotic drug mefloquine against cervical cancer through impairing mitochondrial function and inhibiting mTOR pathway.

  2017cited by this paper
• Lysophagy: A Method for Monitoring Lysosomal Rupture Followed by Autophagy-Dependent Recovery.

  2017cited by this paper
• L-leucyl-L-leucine methyl ester does not release cysteine cathepsins to the cytosol but inactivates them in transiently permeabilized lysosomes

  2017cited by this paper
• Combining 2-deoxy-D-glucose with fenofibrate leads to tumor cell death mediated by simultaneous induction of energy and ER stress

  2016cited by this paper
• New insight into the role of metabolic reprogramming in melanoma cells harboring BRAF mutations.

  2016cited by this paper
• Synergistic Anticancer Action of Lysosomal Membrane Permeabilization and Glycolysis Inhibition*

  2016cited by this paper
• Solamargine triggers cellular necrosis selectively in different types of human melanoma cancer cells through extrinsic lysosomal mitochondrial death pathway

  2016cited by this paper
• Obatoclax kills anaplastic thyroid cancer cells by inducing lysosome neutralization and necrosis

  2016cited by this paper
• Succinate, an intermediate in metabolism, signal transduction, ROS, hypoxia, and tumorigenesis.

  2016cited by this paper
• Tissue-based map of the human proteome

  2015cited by this paper
• Quinoline-based antimalarial drugs: a novel class of autophagy inhibitors.

  2015cited by this paper
• Distinct cathepsins control necrotic cell death mediated by pyroptosis inducers and lysosome-destabilizing agents

  2015cited by this paper
• The bifunctional autophagic flux by 2-deoxyglucose to control survival or growth of prostate cancer cells

  2015cited by this paper
• Glucose metabolism is inhibited by caspases upon the induction of apoptosis

  2014cited by this paper
• A Proteolytic Cascade Controls Lysosome Rupture and Necrotic Cell Death Mediated by Lysosome-Destabilizing Adjuvants

  2014cited by this paper
• The wonders of 2‐deoxy‐d‐glucose

  2014cited by this paper
• A phase I dose-escalation trial of 2-deoxy-d-glucose alone or combined with docetaxel in patients with advanced solid tumors

  2013cited by this paper
• Autophagy sequesters damaged lysosomes to control lysosomal biogenesis and kidney injury

  2013cited by this paper
• Dichloroacetate inhibits aerobic glycolysis in multiple myeloma cells and increases sensitivity to bortezomib

  2013cited by this paper
• Role of lysosome rupture in controlling Nlrp3 signaling and necrotic cell death

  2013cited by this paper
• Cathepsin-mediated Necrosis Controls the Adaptive Immune Response by Th2 (T helper type 2)-associated Adjuvants*

  2013cited by this paper
• Transformation-associated changes in sphingolipid metabolism sensitize cells to lysosomal cell death induced by inhibitors of acid sphingomyelinase.

  2013cited by this paper
• Lysosomal disruption preferentially targets acute myeloid leukemia cells and progenitors.

  2013cited by this paper
• Differential effects of the glycolysis inhibitor 2‐deoxy‐D‐glucose on the activity of pro‐apoptotic agents in metastatic melanoma cells, and induction of a cytoprotective autophagic response

  2012cited by this paper
• TGF-β signaling, activated stromal fibroblasts, and cysteine cathepsins B and L drive the invasive growth of human melanoma cells.

  2012cited by this paper
• Chloroquine-Mediated Lysosomal Dysfunction Enhances the Anticancer Effect of Nutrient Deprivation

  2012cited by this paper
• Shikonin and its analogs inhibit cancer cell glycolysis by targeting tumor pyruvate kinase-M2

  2011cited by this paper
• 2-Deoxy-d-glucose activates autophagy via endoplasmic reticulum stress rather than ATP depletion

  2011cited by this paper
• Metabolic Agents that Enhance ATP can Improve Cognitive Functioning: A Review of the Evidence for Glucose, Oxygen, Pyruvate, Creatine, and L-Carnitine

  2011cited by this paper
• Comparative Metabolic Flux Profiling of Melanoma Cell Lines

  2011cited by this paper
• The structural biochemistry of the superoxide dismutases.

  2010cited by this paper
• A new understanding in the epidemiology of melanoma

  2010cited by this paper
• Lactate dehydrogenase 5 expression in melanoma increases with disease progression and is associated with expression of Bcl-XL and Mcl-1, but not Bcl-2 proteins

  2010cited by this paper
• Targeting tumor metabolism with 2‐deoxyglucose in patients with castrate‐resistant prostate cancer and advanced malignancies

  2010cited by this paper
• p53 Protects lung cancer cells against metabolic stress.

  2010cited by this paper
• Identification of melanoma antigens using a Serological Proteome Approach (SERPA).

  2010cited by this paper
• Oxidative stress-induced necrotic cell death via mitochondira-dependent burst of reactive oxygen species.

  2009cited by this paper
• 2-Deoxy-D-glucose enhances TRAIL-induced apoptosis in human melanoma cells through XBP-1-mediated up-regulation of TRAIL-R2

  2009cited by this paper
• How mitochondria produce reactive oxygen species

  2008cited by this paper
• The normalization of gene expression data in melanoma: investigating the use of glyceraldehyde 3-phosphate dehydrogenase and 18S ribosomal RNA as internal reference genes for quantitative real-time PCR.

  2008cited by this paper
• Cysteine Cathepsins Trigger Caspase-dependent Cell Death through Cleavage of Bid and Antiapoptotic Bcl-2 Homologues*

  2008cited by this paper
• Succinate Ameliorates Energy Deficits and Prevents Dysfunction of Complex I in Injured Renal Proximal Tubular Cells

  2008cited by this paper
• Role of the mitochondrial membrane permeability transition in cell death

  2007cited by this paper
• Vincristine induces dramatic lysosomal changes and sensitizes cancer cells to lysosome-destabilizing siramesine.

  2007cited by this paper

• No citing papers are available for this paper.