mouse b16f10 melanoma cancer cell line  (ATCC)

Journal: Croatian Medical Journal

Article Title: Ornidazole suppresses CD133 + melanoma stem cells via inhibiting hedgehog signaling pathway and inducing multiple death pathways in a mouse model

doi: 10.3325/cmj.2022.63.461

Figure Lengend Snippet: Results of Crystal Violet and MTT assays. (A) Cellular viability was assessed with Crystal Violet staining. Melanoma cancer cells were treated with the denoted concentrations of ornidazole for 48 h. (B-D) Ornidazole inhibits B16F10 melanoma cancer cells viability as determined by MTT assay. Melanoma cancer cells were treated with different concentrations of ornidazole for (B) 24 h, (C) 48 h, (D) 72 h. Results were presented as mean ± standard deviation of three independent experiments. Each group with different concentration of ornidazole was compared with the control group (one-way ANOVA with Dunnet's test for multiple comparisons, * P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: B16F10 mouse melanoma cancer cell line was purchased from ATCC (CRL-6475, Manassas, VA, USA).

Techniques: Staining, MTT Assay, Standard Deviation, Concentration Assay, Control

Journal: Croatian Medical Journal

Article Title: Ornidazole suppresses CD133 + melanoma stem cells via inhibiting hedgehog signaling pathway and inducing multiple death pathways in a mouse model

doi: 10.3325/cmj.2022.63.461

Figure Lengend Snippet: Wound-healing and Comet assays. The results are expressed as percentage of wound closure, and the area measured at time zero was considered 0% (A) Ornidazole inhibits B16F10 melanoma cancer cells migration as determined by wound-healing analysis. (B) Wound closure rates (%) at 8 h and 24 h. Ornidazole (400 μg/mL, 800 μg/mL, 1200 μg/mL) inhibited cell migration by 15%, 60%, and 96%, respectively, in B16F10 cells after 24 hours (one-way ANOVA followed by Bonferroni testing for multiple comparisons. Data are shown as the mean ± standard deviation, P < 0.05) (C) DNA damage as measured by the Comet assay as a result of treatment with different concentrations of ornidazole (400 μg/mL, 800 μg/mL, 1200 μg/mL) for 24, 48, and 72 hours. Olive moment = (tail mean-head mean) x % of DNA in the tail (D) Mean tail moment (μm) represents the damage distribution in the attached cells. The experiment was done in triplicates, and data are expressed as mean ± standard deviation. Each group was compared with the control group (two-way ANOVA with multiple comparisons, Bonferroni test was used for post hoc analysis, *** P < 0.001).

Article Snippet: B16F10 mouse melanoma cancer cell line was purchased from ATCC (CRL-6475, Manassas, VA, USA).

Techniques: Migration, Standard Deviation, Single Cell Gel Electrophoresis, Control

Journal: Croatian Medical Journal

Article Title: Ornidazole suppresses CD133 + melanoma stem cells via inhibiting hedgehog signaling pathway and inducing multiple death pathways in a mouse model

doi: 10.3325/cmj.2022.63.461

Figure Lengend Snippet: (A) Mice that were injected unsorted B16F10 cells and tumors resected from ornidazole-treated and control mice that received unsorted cell injection. (B) Mice that were injected CD133 + B16F10 cells. (C) Mice that were injected CD133 - B16F10 cells and tumors resected from ornidazole-treated and control mice that received CD133 - cell injection. (D) After ornidazole treatment, average tumor volume measurements of the unsorted group (*** P < 0.001), (E) CD133 + (4th day ** P < 0.01, 9th-12th day *** P < 0.001), and (F) CD133 - groups (2nd day * P < 0.5, 9th-12th day *** P < 0.001). All groups were compared with the control group (two-way ANOVA with Bonferroni correction for multiple comparisons).

Article Snippet: B16F10 mouse melanoma cancer cell line was purchased from ATCC (CRL-6475, Manassas, VA, USA).

Techniques: Injection, Control

Journal: Croatian Medical Journal

Article Title: Ornidazole suppresses CD133 + melanoma stem cells via inhibiting hedgehog signaling pathway and inducing multiple death pathways in a mouse model

doi: 10.3325/cmj.2022.63.461

Figure Lengend Snippet: Gene and relative protein expression levels of hedgehog signaling pathway. (A) Effect of ornidazole on hedgehog signaling pathway in unsorted, CD133 + , and CD133 - cells was assessed with real time-polymerase chain reaction. Tumor cells were isolated from mice. Each group was compared with the control group. Data are shown as mean ± standard deviation from three independent experiments (one-way ANOVA with Bonferroni corrections for multiple comparisons, *** P < 0.001, ** P < 0.01 (n = 6/group). (B) The fold-change levels of protein expression in ELISA assay of the B16F10 cells treated with ornidazole. The experiments were performed in duplicate. Each group was compared with the control group. Results are presented as mean ± standard deviation of three independent experiments (two-way ANOVA with Bonferroni testing for multiple comparisons, *** P < 0.001, ** P < 0.01, n = 6 per group). Shh – Sonic hedgehog, PTCH1 – patched-1, SMO – smoothened, GLI1 – glioma-associated oncogene homologue-1, CD133 – also known as AC133 and prominin-1.

Article Snippet: B16F10 mouse melanoma cancer cell line was purchased from ATCC (CRL-6475, Manassas, VA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Control, Standard Deviation, Enzyme-linked Immunosorbent Assay

Journal: Nature Communications

Article Title: Mitochondrial fission induces immunoescape in solid tumors through decreasing MHC-I surface expression

doi: 10.1038/s41467-022-31417-x

Figure Lengend Snippet: A Membrane expression of MHC-I in TSCCs and B16F10 was assessed by flow cytometry with indicated treatments (mean ± s.e.m; n = 6; p = 0.0029, 0.0063 for SCC-9, 0.0037, 0.0002 for CAL-27, 0.001, 0.002 for B16F10; * p < 0.01, ** p < 0.001, siTPP2 compared with NC and siTPP2+XBP-1s, XBP-1s compared with vector and siTPP2+XBP-1s by two-way ANOVA followed by Dunnett’s tests for multiple comparisons). M indicates the mean fluorescence intensity. B SIINFEKL-H-2K b complex expression was evaluated by flow cytometry of B16F10 (mean ± s.e.m; n = 4; p = 0.0113 and p < 0.0001; * p < 0.01, ** p < 0.001 compared with shCtrl by two-tailed t -test). C Scheme of in vitro antigen presentation assays. D Quantification of mouse IFN-γ production by ELISA after 24 h (mean ± s.e.m; n = 5; p = 0.001, p < 0.0001, p < 0.001 for OVA and p < 0.0001 for SIINFEKEL; * p < 0.01, ** p < 0.001 compared with shCtrl by two-tailed t -test). E Twelve hours after coculture, GFP + cancer cells were harvested and cell death was examined by flow cytometry based on the uptake of PI and Annexin V (mean ± s.e.m; n = 4; p = 0.0003, p < 0.0001, p < 0.0001; * p < 0.01, ** p < 0.001 compared with shCtrl by two-tailed t -test). E indicates effector cells namely T cells. T indicates targeted cells namely cancer cells. F Additional transmembrane molecules expression after DRP-1 knockdown was assessed by flow cytometry ( n = 3). Both shCtrl and NC indicate negative control. G Graphic abstract of this study.

Article Snippet: Human cancer cell lines SCC-9 (TSCC), CAL-27 (TSCC), A549 (NSCLC), Saos-2 (osteosarcoma), and mouse cancer cell line B16F10 (melanoma) were purchased from American Type Culture Collection.

Techniques: Membrane, Expressing, Flow Cytometry, Plasmid Preparation, Fluorescence, Two Tailed Test, In Vitro, Immunopeptidomics, Enzyme-linked Immunosorbent Assay, Knockdown, Negative Control