Research Article

Cucurbitacin D Induces G2/M Phase Arrest and Apoptosis via the ROS/p38 Pathway in Capan-1 Pancreatic Cancer Cell Line

Figure 4

Reactive oxygen species (ROS) regulate cucurbitacin D (CuD)-induced cell cycle arrest and apoptosis: (a) cells were treated with various concentrations of CuD (0.05, 0.1, and 0.2 μM) for 24 h followed by 10 μM of 2′,7′-dichlorofluorescein diacetate (DCF-DA) for 30 min. The level of ROS was analyzed using flow cytometry. Fluorescence intensity is presented as mean ± SD from three independent experiments. and , significantly different as compared to the control, (b) Capan-1 cells were preincubated with N-acetyl-L-cysteine (NAC; 5 mM) for 1 h and then treated with CuD (0.1 μM) for 24 h. Fluorescence intensity is presented as mean ± SD values from three independent experiments. , significantly different as compared to the control and NAC-treated groups, (c) cell viability was analyzed using the MTT assay in CuD-treated Capan-1 cells, pretreated with NAC (5 mM). Results are presented as mean ± SD values from three independent experiments. , significantly different as compared to the control, and (d and e) western blotting analysis of G2/M cell cycle arrest-related proteins and apoptosis-related proteins in the CuD-treated Capan-1 cells, pretreated with NAC (5 mM).
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