ROS is not involved in induction of cell death by Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid in HepG2 cells

Objective: To identify the role of reactive oxygen species(ROS) formation on cell death induced by Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid(5F) in HepG2 cells.Method: MTT assay was used to determine the effect of 5F on proliferation of HepG2 cells,and apoptotic morphological changes were assessed using Hoechst/PI assay.To evaluate intracellular ROS levels,a GENMED kit was used.HepG2 cells were treated with 5F for 24 h or with 1 mmol·L-1 GSH for 1 h prior to treatment with 5F for 24 h,then cytoplasmic mono-and oligonucleosomes were assessed with Cell Death Detection ELISA kit.Result: The cytotoxicity of 5F on HepG2 cells was elevated with increasing 5F concentrations,as evidenced by the cell viability assay,and the apoptotic changes such as chromatin condensation were confirmed by Hoechst/PI staining.The decrease in ROS generation was observed in HepG2 cells following treatment with 5F.Cytoplasmic mono-and oligonucleosomes induced by 5F were not changed by decreasing basal level of ROS-mediated signaling with GSH.Farther more,induction of ROS production by cisplatinum(CDDP) was canceled by treatment with 5F and 5F revealed a additive effect to cell killing by CDDP.Conclusion: 5F can not only induce apoptosis through non-ROS-depandent pathway,and can abate oxidant stress.