Journal: Biochemistry and Biophysics Reports

Article Title: Ozonated olive oil inhibits melanoma proliferation by inducing ferroptosis

doi: 10.1016/j.bbrep.2025.102267

Figure Lengend Snippet: Ozonated olive oil has ability to inhibit cell migration and proliferation in melanoma cells (A) Seeded A375 cells were scratched (top panels) and incubated with or without 0.001 % or 0.01 % OZO or OLO for 18 h (bottom panels). Blue lines are shown as borders of seeded cells. The images are representative of 3 independent experiments (n = 3). Scale bars, 200 μm. (B) Proportion of invasion areas in OZO-treated and OLO-treated A375 cells are shown. Data were shown mean + SEM of 3 independent experiments. (C–G) Cell viability of A375 (C), G361 (D), Malme-3M (E), B16F10 (F) and HaCaT (G) cells in the presence of 0.01 % OZO (filled circles) or OLO (open circles). Data were shown mean + SEM of 3 independent experiments (n = 3). Statistical analysis performed by Tukey's multiple comparison test. ∗; p < 0.05, ∗∗∗; p < 0.001 (vs OLO). (−): no addition, D: DMSO-treated.

Article Snippet: Human melanoma cell line, A375 cells, G361 cells and Malme-3M cells, human keratinocyte cell line, HaCaT cells and murine melanoma cell line, B16F10 cells were obtained from ATCC and cultured in Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich Co., St. Louis, MO) contained 10 % fetal bovine serum (FBS; Cosmo Bio Co., Tokyo, Japan) at 37 °C in humidified 5 % CO 2 /95 % air atmosphere.

Techniques: Migration, Incubation, Comparison

Journal: Biochemistry and Biophysics Reports

Article Title: Ozonated olive oil inhibits melanoma proliferation by inducing ferroptosis

doi: 10.1016/j.bbrep.2025.102267

Figure Lengend Snippet: Ozonated olive oil induces ferroptosis in melanoma cells (A) Level of intracellular GSH and GSSG ratio in A375, G361, Malme-3M and HaCaT cells treated with or without 0.01 % OLO or 0.01 % OZO for 6h. Data was shown mean + SEM of 3 independent experiments (n = 3). (B–D) Levels of lipid peroxidation in A375 (B), G361 (C) and Malme-3M (D) cells treated with or without 0.01 % OZO- or 0.01 % OLO analyzed by flowcytometric analysis. (Upper panel) The images are representative of individual independent experiments. (Lower panel) The graphs are shown that measured Geometric MFI from 8 (A375), and 4 (other) independent experiments. Statistical significance by Tukey's multiple comparison test. ∗; p < 0.05, ∗∗∗; p < 0.001. (−): no addition, D: DMSO-treated. (E–F) Expression of GPX4 mRNA and protein in A375 at 0–6h after treatment with 0.01 % OLO or 0.01 % OZO. (E) The graph is shown that fold change of GPX4 mRNA levels. Data was shown mean + SEM of 5 independent experiments (n = 5). (F). Data are representative of 5 independent experiments. (F, right panel) The graph is shown that measured GPX4 and GAPDH protein ratio by Image J from 5 independent experiments (n = 5). Statistical significance by Tukey's multiple comparison test. ns; not significant, ∗∗; p < 0.01 (vs OLO).

Article Snippet: Human melanoma cell line, A375 cells, G361 cells and Malme-3M cells, human keratinocyte cell line, HaCaT cells and murine melanoma cell line, B16F10 cells were obtained from ATCC and cultured in Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich Co., St. Louis, MO) contained 10 % fetal bovine serum (FBS; Cosmo Bio Co., Tokyo, Japan) at 37 °C in humidified 5 % CO 2 /95 % air atmosphere.

Techniques: Comparison, Expressing

Journal: Biochemistry and Biophysics Reports

Article Title: Ozonated olive oil inhibits melanoma proliferation by inducing ferroptosis

doi: 10.1016/j.bbrep.2025.102267

Figure Lengend Snippet: Ferroptosis inhibitors but not apoptosis and necroptosis inhibitor suppressed ozonated olive oil-induced melanoma cell death (A-B) Cell viability of 0.01 % OZO- or 0.01 % OLO-treated A375 cells in the presence or absence of 10 μM α-tocopherol (α-Toco) (A), 1 μM ferrostatin-1 (Fer-1) (B) Data was shown mean + SEM 4 of individual independent experiments (n = 4). ∗∗∗; p < 0.001 by Tukey's multiple comparison test. (C–E) Cell viability of 0.01 % OZO- or 0.01 % OLO-treated G361 (C), Malme-3M (D) and B16F10 (E) cells in the presence or absence of ferrostatin-1 (Fer-1). (F, G) Proliferation of 0.01 % OZO- or OLO-treated A375 (F) and Malme-3M (G) cells in the absence or presence of 100 μM deferiprone (DFP). Data was shown mean + SEM of 4 individual independent experiments (n = 4). (H) Proliferation of 0.003 % OZO- or OLO-treated A375 cell in the absence or presence of 100 μM FeSO 4 . Data was shown mean + SEM of 3 individual independent experiments (n = 3). (I) Proliferation of 0.01 % OZO- or OLO-treated A375 cells in the absence or presence of 10 μM Z-VAD (left panel) or 1 μM necrostatin-1 (Nec-1, right panel). Data was shown mean + SEM of 4 individual independent experiments (n = 4). Statistical significance by Tukey's multiple comparison test. ns; not significant, ∗; p < 0.05, ∗∗; p < 0.01, ∗∗∗; p < 0.001. (−): no addition, D: DMSO-treated.

Article Snippet: Human melanoma cell line, A375 cells, G361 cells and Malme-3M cells, human keratinocyte cell line, HaCaT cells and murine melanoma cell line, B16F10 cells were obtained from ATCC and cultured in Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich Co., St. Louis, MO) contained 10 % fetal bovine serum (FBS; Cosmo Bio Co., Tokyo, Japan) at 37 °C in humidified 5 % CO 2 /95 % air atmosphere.

Techniques: Comparison

Journal: Current Issues in Molecular Biology

Article Title: Resveratrol Targets Glycolytic Enzymes HK II and PKM2 to Promote Concurrent Apoptotic and Necrotic Cell Death in Malignant Melanoma

doi: 10.3390/cimb47121006

Figure Lengend Snippet: Effects of resveratrol on cell viability and morphology in malignant melanoma cells. ( A ) Chemical structure of resveratrol (RSV). ( B ) Cell viabilities of non-tumorigenic human epidermal melanocytes (HEMn-MP) and malignant melanoma cell lines (G361 and SK-MEL-24) after 48 h of RSV treatment, assessed by MTT assay. Results are presented as means ± Standard error (SE) of three independent experiments (* p < 0.05). ( C ) Representative phase-contrast images showing morphological alterations in HEMn-MP, G361, and SK-MEL-24 cells following RSV exposure for 48 h (scale bar = 50 μm; original magnification ×400). RSV: resveratrol.

Article Snippet: Normal human epidermal melanocytes (HEMn-MP) were obtained from Cascade Biologics (Portland, OR, USA), and the human malignant melanoma cell lines G361 and SK-MEL-24 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: MTT Assay

Journal: Current Issues in Molecular Biology

Article Title: Resveratrol Targets Glycolytic Enzymes HK II and PKM2 to Promote Concurrent Apoptotic and Necrotic Cell Death in Malignant Melanoma

doi: 10.3390/cimb47121006

Figure Lengend Snippet: Resveratrol induces apoptotic and necroptotic cell death in malignant melanoma cells. G361 and SK-MEL-24 cells were treated with RSV (20, 40, 60 μM) for 48 h. ( A ) Representative fluorescence images of DAPI-stained nuclei (scale bar = 25 μm; original magnification ×200). Condensed or fragmented nuclei, indicative of apoptosis, are indicated by white arrows. ( B ) Quantification of apoptotic cell populations by Annexin-V/7-AAD staining using a Muse™ Cell Analyzer. Annexin-V-positive/7-AAD-negative cells were classified as early apoptotic, whereas Annexin-V-positive/7-AAD-positive cells were classified as late apoptotic. Data are presented as mean ± SE ( n = 3). Statistical significance: * p < 0.05, ** p < 0.01, **** p < 0.0001. RSV: resveratrol.

Article Snippet: Normal human epidermal melanocytes (HEMn-MP) were obtained from Cascade Biologics (Portland, OR, USA), and the human malignant melanoma cell lines G361 and SK-MEL-24 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Fluorescence, Staining

Journal: Current Issues in Molecular Biology

Article Title: Resveratrol Targets Glycolytic Enzymes HK II and PKM2 to Promote Concurrent Apoptotic and Necrotic Cell Death in Malignant Melanoma

doi: 10.3390/cimb47121006

Figure Lengend Snippet: Resveratrol induced G0/G1 cell cycle arrest in malignant melanoma cells. G361 and SK-MEL-24 cells were treated with RSV (0, 20, 40, 60 µM) for 48 h. ( A ) Representative histograms of cell cycle distribution following staining with Muse™ Cell Cycle Reagent and analysis on a Muse™ Cell Analyzer. ( B ) Quantitative analysis of the percentage of cells in G0/G1, S, and G2/M phases. Data represent mean ± SE ( n = 3). ( C ) Western blot analysis of cell cycle regulatory proteins (cyclin D1, cyclin E1, and CDK4) after 48 h of RSV treatment. β-Actin served as a loading control. Relative band intensities were quantified against the control. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (post hoc test). RSV: resveratrol.

Article Snippet: Normal human epidermal melanocytes (HEMn-MP) were obtained from Cascade Biologics (Portland, OR, USA), and the human malignant melanoma cell lines G361 and SK-MEL-24 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Staining, Western Blot, Control

Journal: Current Issues in Molecular Biology

Article Title: Resveratrol Targets Glycolytic Enzymes HK II and PKM2 to Promote Concurrent Apoptotic and Necrotic Cell Death in Malignant Melanoma

doi: 10.3390/cimb47121006

Figure Lengend Snippet: Resveratrol modulates glycolytic enzymes and induces apoptosis and necroptosis in malignant melanoma cells. G361 and SK-MEL-24 cells were treated with RSV (0, 20, 40, 60 µM) for 48 h. ( A ) Western blot analysis of HK II, PKM2, and apoptosis-related proteins (cleaved caspase-3, caspase-3, Bax, and Bcl-2). β-Actin served as loading control. ( B ) Western blot analysis of necroptosis-associated proteins, including phosphorylated MLKL (p-MLKL) and phosphorylated RIP (p-RIP). Total MLKL and total RIP were used as loading controls. ( C ) Intracellular ATP levels. ( D ) Hexokinase enzymatic activity. ( E ) Caspase-3/7 activity. Data are expressed as mean ± SE ( n = 3). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). HK II: Hexokinase II; PKM2: pyruvate kinase M2; RSV: resveratrol.

Article Snippet: Normal human epidermal melanocytes (HEMn-MP) were obtained from Cascade Biologics (Portland, OR, USA), and the human malignant melanoma cell lines G361 and SK-MEL-24 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Western Blot, Control, Activity Assay

Journal: Current Issues in Molecular Biology

Article Title: Resveratrol Targets Glycolytic Enzymes HK II and PKM2 to Promote Concurrent Apoptotic and Necrotic Cell Death in Malignant Melanoma

doi: 10.3390/cimb47121006

Figure Lengend Snippet: Resveratrol suppresses the migratory ability of malignant melanoma cells. A wound healing assay was performed in G361 ( A ) and SK-MEL-24 ( B ) cells treated with RSV (0, 20, 40, 60 μM) for 48 h. Assays were conducted under both 5% FBS-supplemented and serum-free conditions to minimize confounding effects of proliferation. Wound areas were imaged at 0 h and 48 h to assess cell migration (scale bar = 50 μm; original magnification ×400). Data are presented as mean ± SE ( n = 3). Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (post hoc test). RSV: resveratrol.

Article Snippet: Normal human epidermal melanocytes (HEMn-MP) were obtained from Cascade Biologics (Portland, OR, USA), and the human malignant melanoma cell lines G361 and SK-MEL-24 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Wound Healing Assay, Migration