Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Nrf2 knockdown enhances the reduction in viability of melanoma cells following their exposure to leukoderma-inducing phenolic compounds in a tyrosinase-dependent manner. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Nrf2 (#1, MSS275988) or Tyr (#1, MSS212191) for 24 h. ( A , D ) Levels of Nrf2 and Tyr mRNA following 24 h incubation in control medium. ( B , C , E , F ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data are presented as the means ± SD of independent experiments (n = 3 for ( A ) or as the number indicated for ( C , F )) or the means ± SD (n = 3 wells) with similar results in more than two independent experiments (for ( B , D , E )). *, p < 0.05, versus control siRNA-transfected cells (for ( A , D )) or between respective values. B16BL6 melanoma cells (for G – J ) were transfected with a siRNA directed against Nrf2 (#2, MSS207018), Tyr (#2, MSS212190), or a negative control siRNA ( Ctrl ) for 24 h. B16-4A5 melanoma cells (for ( K – M )) were transfected with Nrf2 (#1) and Tyr (#1). ( G , K , M ). Levels of Nrf2 or Tyr mRNA following 24 h incubation in control medium. ( H , I , J , L ) Viability of the cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments (for ( H , I , J , L )). For M, data represent the means ± SD of independent experiments (as the number indicated). *, p < 0.05, between respective values or versus control siRNA-transfected cells (for ( G , K )).

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Transfection, Negative Control, Incubation, Control, Concentration Assay

Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Nrf2 knockdown reduces GSH , Slc7a11, and Nqo1 mRNA levels and effectively prevents their induction by phenolic compounds. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Nrf2 (#1) for 24 h. The cells were further incubated in the medium containing a vehicle or compounds (0.3 mM) for 24 h. ( A , B ) Cellular GSH levels. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments. ( C , D ) Levels of Slc7a11 , Nqo1 , and Nrf2 mRNA. Data represent means ± SD of independent experiments (n = 3 for ( C ), or as the number indicated for ( D )). *, p < 0.05, versus control siRNA-transfected cells; #, p < 0.05, versus vehicle-treated cells.

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Transfection, Negative Control, Incubation, Control

Journal: Biomolecules

Article Title: A Cell-Based Evaluation of the Tyrosinase-Mediated Metabolic Activation of Leukoderma-Inducing Phenols, II: The Depletion of Nrf2 Augments the Cytotoxic Effect Evoked by Tyrosinase in Melanogenic Cells

doi: 10.3390/biom15010114

Figure Lengend Snippet: Slc7a11 knockdown enhances the cytotoxicity induced by RD, RK, and pCRE, whereas Nqo1 knockdown or inhibition is ineffective for most compounds. B16BL6 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Slc7a11 or Nqo1 for 24 h. ( A ) Slc7a11 and Nqo1 mRNA levels following 24 h incubation in a control medium. ( B – D ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. ( E , F ) The viability of B16BL6 cells after exposure to the indicated concentration of phenolic compounds in the presence or absence of ES936 (500 nM) for 24 h. ( F ) Cells were pretreated with a vehicle or ES936 for 24 h before their exposure to the compounds with/without ES936. ( G ) Western blots and quantitation of tyrosinase and the loading control of β-actin in cells at 48 h after Nqo1 -siRNA transfection or 24 h after ES936 treatment. Data represent means ± SD (n = 3 wells) with similar results in two independent experiments (for ( A – F )) or means ± SD of independent experiments (n = 4 for RD in panel C, D or n = 3 for 4SCAP in panel D). *, p < 0.05, versus Ctrl -siRNA-transfected cells (for ( A )) or between respective values (for ( B – F )). B16-4A5 melanoma cells were transfected with a negative control siRNA ( Ctrl ) or a siRNA directed against Slc7a11 or Nqo1 for 24 h. ( H ) Levels of Slc7a11 and Nqo1 mRNA following 24 h incubation in a control medium. ( I ) The viability of cells following exposure to the indicated concentration of compounds for 24 h or 48 h. Data represent means ± SD (n = 3 wells). Similar results were obtained in two independent experiments (for ( I )). *, p < 0.05, versus Ctrl -siRNA-transfected cells.

Article Snippet: Mouse melanoma cell lines B16BL6 and B16 melanoma 4A5 (B16-4A5) were purchased from the RIKEN BioResource Research Center (Tsukuba, Japan) and maintained in RPMI1640 medium and Dulbecco’s modified Eagle’s medium (DMEM), respectively, containing 10% FBS.

Techniques: Knockdown, Inhibition, Transfection, Negative Control, Incubation, Control, Concentration Assay, Western Blot, Quantitation Assay

Journal: International Journal of Molecular Sciences

Article Title: Organogermanium THGP Induces Differentiation into M1 Macrophages and Suppresses the Proliferation of Melanoma Cells via Phagocytosis

doi: 10.3390/ijms24031885

Figure Lengend Snippet: THGP increases the cytotoxicity of RAW 264.7 cells toward B16 4A5 melanoma cells. ( a ) The cytotoxicity of RAW 264.7 cells against B16 4A5 cells was evaluated via MTS assay. ( b ) The cytotoxicity of RAW 264.7 cells against B16-F10 LUC #2 cells was evaluated using a luciferase assay. ( c ) After staining with the dye CMFPTX, B16 4A5 cells (red) and RAW 264.7 cells (green) were cocultured and analyzed using flow cytometry. ( d ) Then, the percentages of B16 4A5 cells were determined. ( e ) The image shows apoptotic cells analyzed with PI and Hoechst staining. Total cells were stained blue with Hoechst 33258. RAW 264.7 cells were stained green with Cell Tracker Green. Apoptotic cells were stained red with propidium iodide. Apoptotic B16 4A5 cells are shown as blue + green − red + cells (shown by white arrows). Live B16 4A5 cells are shown as blue + green − red − cells. Apoptotic RAW 264.7 cells are shown as blue + green + red + cells. Live RAW 264.7 cells are shown as blue + green + red − cells. Scale bars: 20 μm. ( f ) The graph shows the percentage of apoptotic B16 4A5 cells cocultured with control RAW 264.7 cells or THGP-treated RAW 264.7 cells for 10 days or more. ( g ) The figure shows the growth rate of B16 4A5 cells cultured in conditioned medium supplemented with or without 500 μM THGP and in the mixture of the RAW 264.7 culture supernatant supplemented with or without 500 μM THGP and normal medium at a 1:1 ratio determined using the MTT assay. Ctrl: normal medium; THGP: normal medium containing 500 μM THGP; C-CM: untreated RAW 264.7 cell culture supernatant; T-TM: THGP-treated RAW 264.7 cell culture supernatant. n = 6. * p < 0.05 and ** p < 0.01 compared with the control. N.S.= Not significant.

Article Snippet: After 24 h, B16 4A5 cells were stained with CellTracker RED CMFPTX Dye (Thermo Fisher Scientific Inc.), and 1 × 10 6 cells were seeded on RAW 264.7 cells.

Techniques: MTS Assay, Luciferase, Staining, Flow Cytometry, Cell Culture, MTT Assay

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: The appearance of tumor cells in culture. (A) A375 (human melanoma cells); (B) HepG2 (human liver carcinoma cells); (C) B16-F0 (murine melanoma cells); (D) B164A5 (murine melanoma cells); (E) MCF7 (breast carcinoma cells); and (F) MDA-MB-231 (breast carcinoma cells).

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: The migratory capacity of murine melanoma cells (B16-F0 and B164A5) over a period of 24 h. The contrast of the images was increased for better visualization.

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: PCA spectra clustering using 9,500–12,500 m/z interval. The top left score plot is a 3D plot of PC1 vs. PC2 vs. PC3 while on right is a 2D plots of PC1 vs. PC2, bottom left is PC1 vs. PC3 and bottom right PC2 vs. PC3. Each dot represents one spectra: 1, A375 (human melanoma); 2, HepG2 (human liver carcinoma); 3, B16-F0 (murine melanoma); 4, B164A5 (murine melanoma); 5, MCF7 (human breast carcinoma); and 6, MDA-MB-231 (human breast carcinoma). PCA, principle component analysis.

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: PCA dendogram. 1, A375 (human melanoma); 2, HepG2 (human liver carcinoma); 3, B16-F0 (murine melanoma); 4, B164A5 (murine melanoma); 5, MCF7 (human breast carcinoma); and 6, MDA-MB-231 (human breast carcinoma). PCA, principle component analysis.

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: The classification results obtained by comparing the MSP in rapport with each other.

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Classification of cancer cell lines using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and statistical analysis

doi: 10.3892/ijmm.2017.3083

Figure Lengend Snippet: The MSP dendrogram. 1, A375 (human melanoma); 2, HepG2 (human liver carcinoma); 3, B16-F0 (murine melanoma); 4, B164A5 (murine melanoma); 5, MCF7 (human breast carcinoma); and 6, MDA-MB-231 (human breast carcinoma). MSP, main spectra.

Article Snippet: The cancer cell lines used in the present study, B16-F0 [murine melanoma; CRL-6322™, American Type Culture Collection (ATCC), Manassas, VA, USA], B16 melanoma 4A5 (murine melanoma; 94042254; Sigma-Aldrich Chemie GmbH, Munich, Germany), A375 (human melanoma; CRL-1619™; ATCC), HepG2 (human liver carcinoma; HB8065™; ATCC), MCF7 (human breast carcinoma; HTB22™; ATCC) and MDA-MB-231 (human breast carcinoma; HTB26™; ATCC), were acquired from Sigma-Aldrich Chemie GmbH and ATCC as frozen items.

Techniques: