malignant melanoma cell line g361  (ATCC)

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

Journal: Annals of dermatology

Article Title: Overexpression of Dock180 and Elmo1 in Melanoma is Associated with Cell Survival and Migration.

doi: 10.5021/ad.23.023

Figure Lengend Snippet: Fig. 2. Apoptotic effects following knockdown of Dock180 and Elmo1 in G361 and SK-MEL-2 cells. Cells were treated with Dock180- or Elmo1-specific small interfering RNA (siRNA). (A) The percentage of cell viability was measured by MTT assay as mean±standard devia- tions for three independent experiments. (B) Phase contrast images of G361 and SK-MEL-2 cells. (C) In DAPI staining, both chromatin con- densation and nuclear fragmentation were increased in G361 and SK-MEL-2 cells following treatment of si-Dock180 or si-Elmo1. si-Ctrl: siRNA control, si-Dock180: siRNA targeting Dock180, si-Elmo1: siRNA targeting Elmo1. *p<0.05 compared to untreated controls.

Article Snippet: The human melanoma cell lines G361 and SK-MEL-2 were purchased from the American Type Culture Collection (CRL- 1424 and HTB-68TM; ATCC).

Techniques: Knockdown, Small Interfering RNA, MTT Assay, Staining, Control

Journal: Annals of dermatology

Article Title: Overexpression of Dock180 and Elmo1 in Melanoma is Associated with Cell Survival and Migration.

doi: 10.5021/ad.23.023

Figure Lengend Snippet: Fig. 4. Inhibition of Dock180 and Elmo1 changes cell cycles and apoptosis of G361 and SK-MEL-2 cells. (A) Cell distribution at G0/G1, S, and G2/M phases was analyzed at 48 hours of incubation time using flow cytometry after staining with propidium iodide (20 μg/ml). (B) The percentage of apoptotic cells after Annexin V-PE binding was analyzed at 48 hours of incubation time using a Muse cell analyzer. The quantitative data were shown as mean±standard deviations for three independent experiments. si-Ctrl: siRNA control, si-Dock180: siRNA targeting Dock180, si-Elmo1: siRNA targeting Elmo1. *p<0.05 compared to untreated controls.

Article Snippet: The human melanoma cell lines G361 and SK-MEL-2 were purchased from the American Type Culture Collection (CRL- 1424 and HTB-68TM; ATCC).

Techniques: Inhibition, Incubation, Flow Cytometry, Staining, Binding Assay, Control

Journal: Annals of dermatology

Article Title: Overexpression of Dock180 and Elmo1 in Melanoma is Associated with Cell Survival and Migration.

doi: 10.5021/ad.23.023

Figure Lengend Snippet: Fig. 5. Inhibition of migration and colony formation in G361 and SK-MEL-2 cells by knockdown of Dock180 and Elmo1. (A) Transfec- tion with Dock180- or Elmo1-specific small interfering RNA (siRNA) induced the inhibition of melanoma cell migration. At 24 hours post-transfection, the cells were grown to near confluence, and then wounded by dragging a 20 µl pipette tip through the monolayer. Cell migration images were captured soon after the wound was introduced (0 hour) and at a designated time (48 hours after wounding) under a microscope. (B) In 2 weeks after transfection with Dock180- or Elmo1-specific siRNA, colony formation of the G361 cells was suppressed. Immunoblot analysis of Dock180, Elmo1, Rac1 and phosphorylation of ERK and AKT in G361 and SK-MEL-2 cells treated with Dock180- or Elmo1-specific siRNA. si-Ctrl: siRNA control, si-Dock180: siRNA targeting Dock180, si-Elmo1: siRNA targeting Elmo1. *p<0.05 compared to untreated controls.

Article Snippet: The human melanoma cell lines G361 and SK-MEL-2 were purchased from the American Type Culture Collection (CRL- 1424 and HTB-68TM; ATCC).

Techniques: Inhibition, Migration, Knockdown, Small Interfering RNA, Transfection, Transferring, Microscopy, Western Blot, Phospho-proteomics, Control