Journal: Oxidative Medicine and Cellular Longevity

Article Title: Apigenin Induced Apoptosis by Downregulating Sulfiredoxin Expression in Cutaneous Squamous Cell Carcinoma

doi: 10.1155/2022/8172866

Figure Lengend Snippet: Apigenin can downregulate Srx expression in TPA-induced JB6 and A431 cSCC cells. (a) JB6 was treated with control (DMSO), TPA (20 nM), or apigenin (40 or 80 μ M) for different time points (6 h-48 h). Srx expression was detected by western blotting (left). Western blot was served to analyze the expression of Srx while TPA-induced JB6 cells were incubated with apigenin (80 μ M) for different times (6 h-48 h) (right). (b) Human SCC A431 cells were treated with apigenin at different times, and the expression of Srx was detected by WB. (c) The mRNA level of Srx in TPA-induced JB6 and A431 cells was conducted to measure after incubation with 80 μ M apigenin for 6 h and 12 h. (d) Representative images of immunofluorescence staining of Srx in TPA-induced JB6 treated with control (DMSO) or 80 μ M apigenin for 6 h-48 h (left). Quantitative analysis of Srx means fluorescence intensity (MFI) (right) (mean values ± SEM, n = 3). Significant differences were evaluated using a one-way ANOVA. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. control. For JB6 cells, TPA-induced sample as control.

Article Snippet: The mouse epithelial-derived JB6 Cl 41-5a cell was purchased from the American Type Culture Collection (ATCC) and stored in our laboratory [ ].

Techniques: Expressing, Control, Western Blot, Incubation, Immunofluorescence, Staining, Fluorescence

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Apigenin Induced Apoptosis by Downregulating Sulfiredoxin Expression in Cutaneous Squamous Cell Carcinoma

doi: 10.1155/2022/8172866

Figure Lengend Snippet: Apigenin can attenuate cell proliferation and migration in cSCC cells. (a) CCK-8 assay was implied to analyze the cell viability after the treatment of TPA-induced JB6 and A431 cells with different concentrations of apigenin (up to 80 μ M) as time gone (mean values ± SEM, n = 6). ∗∗∗∗ p < 0.0001 vs. control by ANOVA. (b) Representative images of colony formation assay in TPA-induced JB6 treated with control (DMSO) or 80 μ M apigenin. (c) Typical pictures (left) and quantitative analysis (right) of wound healing assay in TPA-induced JB6 (up) and A431 (down) cells. Cells were treated with control (DMSO) or 80 μ M apigenin for indicated time points. ns: no statistical significance; ∗∗∗∗ p < 0.0001 vs. control by Student's unpaired t -test.

Article Snippet: The mouse epithelial-derived JB6 Cl 41-5a cell was purchased from the American Type Culture Collection (ATCC) and stored in our laboratory [ ].

Techniques: Migration, CCK-8 Assay, Control, Colony Assay, Wound Healing Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Apigenin Induced Apoptosis by Downregulating Sulfiredoxin Expression in Cutaneous Squamous Cell Carcinoma

doi: 10.1155/2022/8172866

Figure Lengend Snippet: Apigenin induced apoptosis in cSCC cells. (a) Flow cytometry was used to analyze the apoptosis cells. Cells were stained with annexin V and PI to quantify the percentage of apoptotic cells. TPA-induced JB6 cells (upper) or A431 cells (lower) were treated with control (DMSO) or 80 μ M apigenin for different times. (b) A concrete percentage of apoptosis cells in TPA-induced JB6 and A431 cells were evaluated using a one-way ANOVA (mean values ± SEM, n = 3) ∗ p < 0.05, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. control (TPA-induced sample as control for JB6 cells). (c) TPA-induced JB6 cells were incubated with control (DMSO) or 80 μ M apigenin for 6 h-48 h. Western blot was served to analyze the expression of apoptosis-associated proteins (left). The bar graphs on the right showed the intensity of the protein band from each treatment relative to the housekeeping protein ( β -actin). Valued represent the means ± SEM. Significant difference was designed by ANOVA, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. control (only TPA-induced sample). (d) The apoptosis-associated proteins were detected while 80 μ M apigenin was used for treatment for different time-points in A431 cells. The bar graphs showed the intensity quantification of the protein band relative to the housekeeping protein. Significant difference from control by ANOVA, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001.

Article Snippet: The mouse epithelial-derived JB6 Cl 41-5a cell was purchased from the American Type Culture Collection (ATCC) and stored in our laboratory [ ].

Techniques: Flow Cytometry, Staining, Control, Incubation, Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Apigenin Induced Apoptosis by Downregulating Sulfiredoxin Expression in Cutaneous Squamous Cell Carcinoma

doi: 10.1155/2022/8172866

Figure Lengend Snippet: Apigenin activated the MAPK signaling pathway in cSCC in vitro. (a, b) Variation of the MAPK signaling pathway with the treatment of apigenin in TPA-induced JB6 (a) and A431 cells (b). The cells were treated with 80 μ M apigenin for different time points up to 24 h. Western blot was applied to analyze the expression of MAPK pathway-associated proteins, includingp38, ERK1/2 and JNK compared with GAPDH (left). The bar graph on the right showed the intensity of the phosphorylation protein band from each treatment relative to the total protein. Valued represent the means ± SEM. Significant difference was designed by ANOVA, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. control (TPA-induced sample as control for JB6 cells). (c) TPA-induced JB6 cell was incubated with control (DMSO, TPA-alone, and apigenin-alone) or apigenin (40 or 80 μ M) for indicted time points. Western blot was served to analyze the expression of Nrf2. β -Actin was used as the reference for the loading quantity of protein sample. The bar graph indicated the density quantification of the Nrf2 band relative to β -actin. ∗ p < 0.05 and ∗∗∗ p < 0.001 vs. TPA-induced control by ANOVA.

Article Snippet: The mouse epithelial-derived JB6 Cl 41-5a cell was purchased from the American Type Culture Collection (ATCC) and stored in our laboratory [ ].

Techniques: In Vitro, Western Blot, Expressing, Phospho-proteomics, Control, Incubation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Apigenin Induced Apoptosis by Downregulating Sulfiredoxin Expression in Cutaneous Squamous Cell Carcinoma

doi: 10.1155/2022/8172866

Figure Lengend Snippet: Apigenin induced cell apoptosis and inhibited the expression of Srx via regulating the MAPK signaling pathway in cSCC. (a, b) TPA-induced JB6 cells (a) and A431 cells (b) were treated with the combination of apigenin and inhibitor of MEK1/2, Binimetinib (5 or 10 nM), for 8 h. Western blotting was used to analyze the expression of Srx, p-Erk, and the apoptosis-related proteins BAX and Bcl2. Representative images are shown on the left. The bar graph on the right indicated the intensity quantification of the protein band relative to GAPDH or total protein (ERK). Significant difference was designed by ANOVA, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 vs. control (TPA-induced sample as control for JB6 cells). (c) Real-time PCR was conducted to quantify the mRNA expression of Srx in cSCC cell lines while cells were treated for indicated compounds. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs control by ANOVA analysis. (d) Representative images of cell apoptosis analysis by flow cytometry after 5 nM Binimetinib (MEK1/2 inhibitor) treatment for 24 h, respectively, in TPA-induced JB6 (up) and A431 (down). The percentage of apoptosis cells after apigenin treatment with or without Binimetinib is shown in the bar graph. ∗∗∗∗ p < 0.0001 vs. control by ANOVA analysis. (e, f) TPA-induced JB6 (e) and A431 (f) were incubated with or without the MEK1/2 inhibitor (5 nM Binimetinib) for 24 h in the presence of 80 μ M apigenin. Whole-cell lysates were subjected to western blotting to detect the apoptosis-associated proteins caspase 3, caspase 8, and PARP. The bar graph showed the intensity quantification of the protein bands from each treatment. Significant difference was designed by ANOVA, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001 vs. the apigenin-alone group.

Article Snippet: The mouse epithelial-derived JB6 Cl 41-5a cell was purchased from the American Type Culture Collection (ATCC) and stored in our laboratory [ ].

Techniques: Expressing, Western Blot, Control, Real-time Polymerase Chain Reaction, Flow Cytometry, Incubation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Potent USP10/13 antagonist spautin‐1 suppresses melanoma growth via ROS‐mediated DNA damage and exhibits synergy with cisplatin

doi: 10.1111/jcmm.15093

Figure Lengend Snippet: Spautin‐1 significantly attenuated proliferation of melanoma cells in a time‐ and dose‐dependent manner. A, Viability analysis after the treatment of A375 and Sk‐Mel‐28 cells with different concentration of spautin‐1 (up to 20 µmol/L) for 0, 24, 48 and 72 h. (Mean values ± SEM, n = 6). B, The IC50 values of spautin‐1 in A375 and Sk‐Mel‐28 were automatically calculated by GraphPad Prism software. C, spautin‐1 inhibited A375 and Sk‐Mel‐28 cell colony formation. The number of colonies normalized to the corresponding untreated cells formed colonies. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control group, # P < .05 vs 2.5 µmol/L. D, Non‐tumorigenic cell lines (HaCaT and JB6) were treated with various concentration of spautin‐1 for 72 h as indicated, and cell viability in the presence of HaCat and JB6 was measured by MTS as described in the Section and analysed by GraphPad Prism software (mean values ± SEM, n = 6)

Article Snippet: The human malignant melanoma cell lines (A375 and SK‐Mel‐28), the human keratinocyte HaCaT and the mouse epidermis‐derived JB6 Cl 41‐5a cells were purchased from the American Type Culture Collection (ATCC).

Techniques: Concentration Assay, Software, Control