Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Chemical structures of deguelin, tephrosin and rotenone.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques:

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Tephrosin induced apoptosis of pancreatic cancer cells. ( A ) Cell apoptosis was detected by Hoechst 33258 staining. Cells with condensed or fragmented nuclei after treatment of tephrosin were identified as apoptotic cells. ( B ) Apoptosis of PANC-1 and SW1990 cells were detected by Annexin-V/PI co-staining followed with flow cytometric analysis. ( C ) Effects of the caspase inhibitor Z-VAD-FMK (25 µM) preatment on tephrosin-induced apoptosis in PANC-1 and SW1990 cells. ( D ) LDH was detected to evaluate the apoptotic response to treatment of tephrosin in PANC-1 and SW1990 cells. ( E ) Caspase-3 activity increased after treatment with tephrosin at indicated concentrations for 12 h. ( F ) Treatment of tephrosin up-regulated the expresssion of clevaed caspase-3, caspase-9, and PARP. Data represented as the mean ± SD (standard deviation) of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Staining, Activity Assay, Standard Deviation, Control

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Tephrosin caused ROS generation in human pancreatic cancer cells. ( A ) The levels of ROS in PANC-1 and SW1990 cells were measured by DCF-DA staining 1 h after treatment of tephrosin (1 or 4 µM). The green fluorescence intensity is associated with the degree of ROS. ( B, C ) Intracellular ROS accumulated in a dose- and time-dependent manner in tephrosin-treated PANC-1 and SW1990 cells. ( D ) Tephrosin caused a depletion of intracellular levels of GSH. Data represented as the mean ± SD (standard deviation) of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Staining, Fluorescence, Standard Deviation, Control

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Tephrosin induced apoptosis in PANC-1 and SW1990 cells was dependent on ROS, pretreatment with antioxidants NAC or GSH for 1 h prior to tephrosin exposure prevented tephrosin-induced ROS accumulation, proliferation, and caspase-3 activation. ( A ) ROS generation was detected using flow cytometry after staining with DCF-DA. ( B ) Results are expressed as a percentage of relative fluorescence of DCF-DA to the control. The cell viability ( C ) and activity of caspase-3 ( D ) were detected with or without pre-incubation of NAC or GSH. Data represented as the mean ± SD (standard deviation) of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Activation Assay, Flow Cytometry, Staining, Fluorescence, Control, Activity Assay, Incubation, Standard Deviation

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Effects of tephrosin on mitochondria depolarization in PANC-1 and SW1990 cells. ( A ) Cells were pre-incubated with MitoTEMPO (10 µM) for 1 h, followed by incubation with tephrosin. The fluorescence intensity of MitoSOX Red was assessed using flow cytometry. ( B ) The cell viability was detected with MTT method. ( C ) Fluorescence photomicrograph of changes in the MMPs indicated by staining with JC-1. ( D ) Decreased photometric ratio of red to green fluorescence indicated the depolarization of MMP. ( E ) Cells with different treatment were stained with JC-1, followed by flow cytometric analysis. ( F ) The release of cytochrome c from the mitochondria was detected by using Western blot. Data represented as the mean ± SD (standard deviation) of three independent experiments, *** p < 0.001.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Incubation, Fluorescence, Flow Cytometry, Staining, Western Blot, Standard Deviation

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Tephrosin-induced ROS caused DNA damage in PANC-1 and SW1990 cells. ( A ) Tephrosin induced DNA stand breaks. PANC-1 and SW1990 cells were treated with tephrosin for 24 h, followed by alkaline comet assay. ( B ) Comet assay results are graphed as means ± SD of percent tail DNA from 20 cells at 3 independent gels. ( C ) Cells were treated with tephrosin for different time points, and the expression of p-H2A.X was detected using Western blot. ( E-H ) Cells were pre-incubated with NAC or MitoTEMPO for 1 h, followed by tephrosin treatment for 24 h. The expression of of p-H2A.X was detected using Western blot. Data represented as the mean ± SD (standard deviation) of three independent experiments. * p < 0.05, *** p < 0.001.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Alkaline Single Cell Gel Electrophoresis, Single Cell Gel Electrophoresis, Expressing, Western Blot, Incubation, Standard Deviation

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: Tephrosin suppressed tumor growth in PANC-1 xenografted nude mice. ( A ) Schematic diagram displaying the time course of treatment in mice. ( B ) Representative tumor images from each group after treatment. ( C ) Treatment of tephrosin resulted in significant lower tumor weight compared with control (n =6). ( D ) Tephrosin treatment significantly inhibited the pancreatic cancer cell growth in nude mice. ( E ) No obvious body weight changes of mice between the control and tephrosin-treated groups was observed (n =6). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Control

Journal: Journal of Cancer

Article Title: Tephrosin induces apoptosis of human pancreatic cancer cells through the generation of reactive oxygen species

doi: 10.7150/jca.50360

Figure Lengend Snippet: H&E staining of the important tissues in each group. The mice were treated with tephrosin or control for two weeks, after sacrificing the mice, the organs were taken out for pathological analysis.

Article Snippet: Deguelin, tephrosin and rotenone were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Staining, Control