Journal: International Journal of Molecular Sciences

Article Title: Deoxyshikonin Mediates Heme Oxygenase-1 Induction and Apoptotic Response via p38 Signaling in Tongue Cancer Cell Lines

doi: 10.3390/ijms23137115

Figure Lengend Snippet: Profiling of DSK-induced apoptotic proteome in tongue cancer. ( A ) Representative images of array membranes corresponding to DSK-untreated and -treated protein expression in HSC-3 cells. Differentially expressed proteins are labeled and selected for further verification. ( B ) HSC-3 and SCC-9 cells were treated with various concentrations of DSK for 24 h and assessed for the expression of indicated apoptosis related proteins by immunoblotting. Densitometric analyses were quantified and normalized with internal controls (β-actin). The values represent the mean ± SD of three independent experiments. * p < 0.05 compared with untreated controls using Student’s t -test.

Article Snippet: Proteome of apoptotic tongue cancer cells in response to DSK treatment was analyzed by using a Proteome Profiler Human Apoptosis Array Kit (R&D Systems, Minneapolis, MN, USA), allowing to assess a panel of 35 key apoptotic proteins simultaneously.

Techniques: Expressing, Labeling, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Galectin-9 Induces Mitochondria-Mediated Apoptosis of Esophageal Cancer In Vitro and In Vivo in a Xenograft Mouse Model

doi: 10.3390/ijms20112634

Figure Lengend Snippet: Gal-9 induces apoptosis of ESCC cells. ( A ) Early apoptosis analysis in KYSE-150 cells 6 h after the administration of Gal-9. The lower left quadrant represents viable cells (red dots), the lower right quadrant early apoptotic cells (green dots), and the upper right quadrant indicates the late apoptotic or necrotic cells (blue dots); ( B ) The transition of Annexin V-positive cell number following treatment with Gal-9 for 1, 6, 12, and 48 h. Annexin V-positive cells were significantly increased at 1 h and 6 h after Gal-9 treatment, compared to the control; ( C ) The activation of caspase-3 was observed after 6 h of treatment with 100 nM Gal-9, as evaluated with the colorimetric assay. No significant change in the activity of caspase-4, -7, -8 and -9 was observed. The error bars represent 95% CI.

Article Snippet: The state of apoptosis was analyzed by Annexin V-biotin apoptosis detection kit (BioVision, Milpitas, CA, USA) following double staining with fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI).

Techniques: Activation Assay, Colorimetric Assay, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Galectin-9 Induces Mitochondria-Mediated Apoptosis of Esophageal Cancer In Vitro and In Vivo in a Xenograft Mouse Model

doi: 10.3390/ijms20112634

Figure Lengend Snippet: Western blotting analysis of apoptosis-related regulators. The lysate of KYSE-150 cells treated with or without 100 nM Gal-9 was analyzed by Western blotting. ( A ) Cleaved caspase-3 level was increased at 6 h after Gal-9 treatment; ( B ) No difference was observed in the total amount of c-Jun NH 2 -terminal kinase (JNK) and p38. Phospho-SAPK/JNK (p-JNK) increased in 6 h, while phospho-p38 MAPK (p-p38) increased in 6 and 24 h ( B ); ( C ) An increase in Smac, HtrA2, and cytochrome c level was observed in the cytoplasmic fraction, but no change in the levels of these proteins was observed in the membrane fraction containing mitochondria.

Article Snippet: The state of apoptosis was analyzed by Annexin V-biotin apoptosis detection kit (BioVision, Milpitas, CA, USA) following double staining with fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI).

Techniques: Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Galectin-9 Induces Mitochondria-Mediated Apoptosis of Esophageal Cancer In Vitro and In Vivo in a Xenograft Mouse Model

doi: 10.3390/ijms20112634

Figure Lengend Snippet: Mitochondria-mediated apoptotic signaling pathway by JNK and p38. Activated JNK promotes the release of cytochrome c (Cyt C), Smac/Diablo (Smac), and HtrA2 from mitochondria via the Bid-Bax–dependent mechanism. Cyt C initiates the activation of the caspase-9–dependent caspase cascade. Smac and HtrA2 inhibit inhibitor of apoptosis protein (IAP) and promotes apoptosis. Furthermore, JNK phosphorylates Bad and its sequestering partner 14–3-3 and promotes Bad-mediated neutralization of Bcl2 family. JNK may also phosphorylate Bcl-2 and inhibit its anti-apoptotic activity. Both JNK and p38 MAPK (p38) inhibit Bcl-2 by phosphorylating Bim. In addition, the activated JNK translocates to the nucleus and activates transcription factors. JNK may promote apoptosis by increasing the expression of pro-apoptotic genes.

Article Snippet: The state of apoptosis was analyzed by Annexin V-biotin apoptosis detection kit (BioVision, Milpitas, CA, USA) following double staining with fluorescein isothiocyanate (FITC)-conjugated Annexin V and propidium iodide (PI).

Techniques: Activation Assay, Neutralization, Activity Assay, Expressing

Journal: Cancer prevention research (Philadelphia, Pa.)

Article Title: Honokiol Inhibits Lung Tumorigenesis through Inhibition of Mitochondrial Function

doi: 10.1158/1940-6207.CAPR-14-0091

Figure Lengend Snippet: Effect of honokiol on cell proliferation, cell cycle progression and apoptosis in vitro and in vivo. A, Relative cell proliferation rate of H226 and H520 cells treated with honokiol at various concentrations. B,. Effect of honokiol on cell cycle arrest. C, Effect of honokiol on cleaved-caspase 3/7 staining. Shown is the caspase-3/7 fluorescence objects count from Incucyte. Data shown are the means ± SD (n = 3 per treatment group) D, Effect of honokiol on cell proliferation and apoptosis in NTCU-induced lung scc model. Lungs harvested from mice on the 26 weeks in NTCU study (n = 5 mice/group) were fixed, and the FFPE slides were stained using specific antibodies as detailed in the Materials and Methods section. I, II: Representative picture from immunohistochemistry for Ki-67 (I, control group; II, Honokiol group); III: proliferation index as determined by Ki-67; IV, V: Representative picture from immunohistochemistry for cleaved caspase-3 (III, control group; IV, Honokiol group). VI, apoptotic index as measured by cleaved caspase-3. ***, P < 0.001, control group versus honokiol group.

Article Snippet: Caspase3/7 apoptosis assay Honokiol induced apoptosis was analyzed by measuring caspase-3 activation using the CellPlayer Apoptosis Caspase-3/7 kit (Essen Bioscience) by following manufacturer’s instructions.

Techniques: In Vitro, In Vivo, Staining, Fluorescence, Immunohistochemistry