Yun Feng*, Honghui Huang*, Jing Gao*, Shizhu Liu**, #
*Department of Pharmacology, School of Medicine, Jiangsu University, Zhenjiang, PR China - **Department of Health Management, School of Management, Jiangsu University, Zhenjiang, PR China
Objective: To investigate the mechanism of cell cycle arrest of glioma by salinomycin through ROS-mediated DNA damage and AKI inactivation.
Methods: U87 and U251 glioma cells were cultured in vitro and treated with different concentrations of salamphenicol (0, 1, 2, 4, 8 μM) for 48 h. The cell morphology was observed under inverted microscope, the cell activity was detected by tetrazolium bromide (MTT) assay, the (ROS) level of intracellular active oxygen cluster was detected by DCFH-DA assay, the cell cycle distribution was analysed by flow cytometry, and ROS and superoxide were observed by fluorescence microscope. The expression of specific proteins in U87 human glioma mice was established by Western imprinting, and the expression of protein in U87 human glioma mice was detected by Western imprinting.
Results: MTT results showed that salinomycin significantly inhibited the activities of U87 and U251 cells in a dose-dependent manner. Flow cytometry results showed that salinomycin mainly induced and inhibited the cell cycle of U87 cells at S phase and U251 cells at G1 phase. Western blot analysis showed that CyclinA and CDK-2 were inhibited in U87 cells and CyclinB1 and CyclinD1 were inhibited in U251 cells. Under the action of salinomycin, intracellular ROS content, histone H2A, and phosphorylated p53 protein expression were increased. The results of animal experiments showed that the volume and weight of tumour specimens in mice were inhibited by salinomycin, which activated Ser139-H2A and Ser15-p53 and inhibited the expression of CyclinA and AKT.
Conclusion: Salinomycin can inhibit glioma growth in vivo through ROS-mediated DNA damage and AKT inactivation, leading to cell cycle arrest.
Salinomycin, ROS, DNA damage, AKI deactivation, cell cycle arrest, glioma.
10.19193/0393-6384_2020_1_37