Journal: Biomedicines

Article Title: Ilimaquinone Induces Apoptosis and Autophagy in Human Oral Squamous Cell Carcinoma Cells

doi: 10.3390/biomedicines8090296

Figure Lengend Snippet: Effect of ilimaquinone (IQ) on the viability of oral cancer cells (SCC4 and SCC2095). ( A ) SCC4 and ( B ) SCC2095. The cells were seeded onto 96-well plates, incubated for 24 (■) and 48 h (□), treated with IQ for 48 h and then cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Points, means; bars, S.D. ( n = 3–6). * p < 0.05 and ** p < 0.01 vs. the control group.

Article Snippet: SCC4 human oral squamous cell carcinoma cells were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan), and SCC2095 human oral squamous cell carcinoma cells were kindly provided by Professor Susan R. Mallery (The Ohio State University).

Techniques: Incubation, Control

Journal: Biomedicines

Article Title: Ilimaquinone Induces Apoptosis and Autophagy in Human Oral Squamous Cell Carcinoma Cells

doi: 10.3390/biomedicines8090296

Figure Lengend Snippet: Effect of ilimaquinone (IQ) treatment on apoptosis. ( A ) The percentage of apoptotic cells (Q2 + Q4) after dimethyl sulfoxide (DMSO) vehicle, IQ, or 50 nM staurosporine (Stauro.) treatment for 48 h. SCC4 cells were treated with either DMSO, IQ, or Stauro. in 5% fetal bovine serum (FBS)-supplemented DMEM/F12 medium for 48 h and stained with propidium iodide (PI)/annexin V. Columns, mean; bars, S.D. ( n = 4). ** p < 0.01 vs. the control group. ( B ) Concentration-dependent effect of IQ on caspase-3 activation. Staurosporine (Stauro.; 25 nM) was used as the positive control. Columns, mean; bars, S.D. ( n = 3). * p < 0.05 and ** p < 0.01 vs. the control group. ( C ) Expression of PARP, cleavage caspase-9, and procaspase-8 in SCC4 cells after 48 h treatment in 5% FBS-supplemented DMEM/F12 medium.

Article Snippet: SCC4 human oral squamous cell carcinoma cells were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan), and SCC2095 human oral squamous cell carcinoma cells were kindly provided by Professor Susan R. Mallery (The Ohio State University).

Techniques: Staining, Control, Concentration Assay, Activation Assay, Positive Control, Expressing

Journal: Biomedicines

Article Title: Ilimaquinone Induces Apoptosis and Autophagy in Human Oral Squamous Cell Carcinoma Cells

doi: 10.3390/biomedicines8090296

Figure Lengend Snippet: Effect of IQ on various biomarkers in SCC4 cells. ( A ) Phosphorylation/expression of Akt, p38, Bax, Mcl-1, Bcl-2, and survivin in SCC4 cells. Cells were treated with IQ for 48 h, and cell lysates were immunoblotted as described in Materials and Methods. ( B ) Expression of p-p53 and p53 of IQ in SCC4 cells. ( C ) Western blotting of p-p53 and p53 in SCC4 cells transiently transfected with a control vector or p53 shRNA for 24 h. ( D ) IQ-induced cell viability in p53 knockdown SCC4 cells. After incubation, cells were analyzed using the MTT assays. Columns, mean; bars, S.D. ** p < 0.01.

Article Snippet: SCC4 human oral squamous cell carcinoma cells were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan), and SCC2095 human oral squamous cell carcinoma cells were kindly provided by Professor Susan R. Mallery (The Ohio State University).

Techniques: Phospho-proteomics, Expressing, Western Blot, Transfection, Control, Plasmid Preparation, shRNA, Knockdown, Incubation

Journal: Biomedicines

Article Title: Ilimaquinone Induces Apoptosis and Autophagy in Human Oral Squamous Cell Carcinoma Cells

doi: 10.3390/biomedicines8090296

Figure Lengend Snippet: IQ increases reactive oxygen species (ROS) production. ( A ) Statistical analysis of ROS generation in SCC4 cells after the treatment with only 5 μM of IQ for 3 h or in combination with 5 mM of N -acetylcysteine (NAC) or 5 mM of glutathione (GSH). Cells were stained with carboxy-DCF-DA and analyzed using flow cytometry. Data are expressed as mean ± S.D. ( n = 3). * p < 0.05 and ** p < 0.01. ( B ) IQ-induced H2AX phosphorylation/expression in SCC4 cells.

Article Snippet: SCC4 human oral squamous cell carcinoma cells were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan), and SCC2095 human oral squamous cell carcinoma cells were kindly provided by Professor Susan R. Mallery (The Ohio State University).

Techniques: Staining, Flow Cytometry, Phospho-proteomics, Expressing

Journal: Biomedicines

Article Title: Ilimaquinone Induces Apoptosis and Autophagy in Human Oral Squamous Cell Carcinoma Cells

doi: 10.3390/biomedicines8090296

Figure Lengend Snippet: Ilimaquinone (IQ) induces autophagy. ( A ) Left panel, SCC4 cells were treated with DMSO vehicle, IQ, or rapamycin (RAP) for 24 h, harvested and stained with acridine orange, and examined under a fluorescence microscope; arrows, acidic vesicular organelles (AVOs). Right panel, statistical analysis of AVOs in IQ-treated SCC4 cells. Columns, mean; bars, S.D. ( n = 3). * p < 0.05 and ** p < 0.01 vs. the control group. ( B ) LC3B-II and Atg5 expression in SCC4 cells after IQ treatment for 48 h. ( C ) SCC4 cells were pretreated 20 μM of 3-methyladenine (3-MA) or 20 μM of chloroquine (CQ) for 15 min, followed by incubation with 10 μM IQ for 48 h and dual staining with propidium iodide (PI)/annexin V- fluorescein isothiocyanate (FITC). Percentages in the graphs are representative of cell percentage in the respective quadrants ( n = 3). Columns, means; bars, S.D. * p < 0.05 and ** p < 0.01.

Article Snippet: SCC4 human oral squamous cell carcinoma cells were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan), and SCC2095 human oral squamous cell carcinoma cells were kindly provided by Professor Susan R. Mallery (The Ohio State University).

Techniques: Staining, Fluorescence, Microscopy, Control, Expressing, Incubation