Mechanism of Solamargine-Induced Apoptosis in Glioblastoma U251 Cells
Objective To investigate the effect and mechanism of solamargine(SM)in inducing apoptosis of glioblastoma U251 cells.Methods U251 cells were treated with SM(0,2,4,6,8 µmol/L)for 24 h,which were taken as the solvent control group and SM 1,2,3,and 4 groups respectively.CCK-8 assay and clone formation experiment were used to detect cell viability and proliferation ability respectively,and the cell survival rate and clone formation rate were calculated.Annexin V-FITC/PI dual staining method and flow cytometry were used to detect cell apoptosis,and the apoptosis rate was calculated.The expression level of apoptosis-related proteins was detected by the Western blot.The reactive oxygen species levels and mitochondrial membrane potential levels in U251 cells were observed by 2',7'-Dichlorofluorescein diacetate(DCFH-DA)and JC-1 fluorescent probes respectively.Results Compared with those in the solvent control group,the survival rate and clone formation rate of U251 cells showed a decreased trend with the increase of SM concentration(P＜0.05),while the apoptosis rate of U251 cells showed a increased trend with the increase of SM concentration(P＜0.01).After SM treatment,the expression levels of cell cleaved-poly adenosine diphosphate ribose polymerase(C-PARP),and cleaved-caspase 3(C-caspase 3)protein,and intracellular reactive oxygen species levels showed a increased trend(P＜0.05),and mitochondrial membrane potential showed a decreased trend(P＜0.01).Conclusion SM may inhibit the activity of glioblastoma U251 cells by activating the mitochondrial-mediated apoptosis pathway.
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