Effects Of Casticin On Apoptosis And Mitochondrial Pathway In Cervical Cancer Cells

Objective:The purpose of this study was to examine the apoptotic activity of casticin in human cervical cancer cells and to investigative its molecular mechanism whether was associated with the generation of reactive oxygen species(ROS).Methods:Apoptotic activities of casticin on human cervical cancer HeLa and peripheral blood mononuclear cells (PBMCs) was performed using histone/DNA ELISA assay, flow cytometry with propidium iodide(PI) staining and DNA agarose gel electrophoresis. Caspase activities were assayed using caspase colorimetric activity assay kit.The mitochondrial membrane potential and ROS production was evaluated by flow cytometry analysis after Rh123and DCFH-DA staining. Protein expression of cytochrome c, Bax, Bcl-2, Bcl-xL and XIAP was analyzed by Western blot.Results:The treatment of Casticin caused the fragment of histone/DNA, Sub-G1population accumulation and ROS production in HeLa cells, but not in PBMCs. Furthermore there were typical bands in DNA agarose gel electrophoresis. Apoptosis of HeLa cells induced by casticin was via an activation of mitochondrial release of cytochrome c through mitochondrial transmembrane potential reduction, activation of caspase-3and-9, production of reactive oxygen species. The pan caspase inhibitor zVAD-FMK, the caspase-9inhibitor zLEHD-fmk and N-acetylcysteine suppressed casticin-induced these apoptosis indexs. Bax was up regulated and there was down regulation of Bcl-xL and XIAP expression. However, the expression of Bcl-2showed no change in such same treatment.Conclusion:1. These results indicate that casticin induced apoptosis of cervical cancer HeLa cells in dose-and time-dependent manner.2. Mitochondrial apoptotic pathway was activated by casticin in cervical cancer HeLa cells.3. Casticin in dose-dependent promoted generation of the ROS in cervical cancer HeLa cells..4. Casticin could activate mitochondrial apoptotic pathway mediated by ROS generation...