Cisplatin induces apoptosis of A549 cells by downregulating peroxidase V

X. Chen, K.-W. Wang, Y.-Q. Chen

Medical College, Shandong University, Jinan, China. 181567778@qq.com

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• Oncology

OBJECTIVE: The purpose of the study was to investigate the role of peroxidase V (Prx V) in Cisplatin-induced apoptosis of A549 cells and its underlying mechanism.

MATERIALS AND METHODS: MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay was conducted to evaluate the regulatory effect of Cisplatin on the survival of A549 cells. ROS (Reactive Oxygen Species) level in A549 cells induced with 0, 2, 4, and 6 mol/L Cisplatin for 24 h was determined using immunofluorescence. Apoptosis of Cisplatin-induced A549 cells was determined by immunofluorescence and flow cytometry, respectively. Western blot was performed to detect protein levels of Prx V, Bcl-2 (B-cell lymphoma 2), BAD ,and caspase-3 in Cisplatin-induced A549 cells.

RESULTS: Survival rate of A549 cells gradually decreased with the increased dose of Cisplatin. Immunofluorescence results elucidated that cellular ROS level in Cisplatin-induced A549 cells increases in a dose-dependent manner. Both immunofluorescence and flow cytometry results revealed that the apoptotic rate of A549 cells increases with the elevation of Cisplatin dose. Besides, the apoptotic rate and ROS level of A549 cells were reduced by NAC pretreatment. Western blot results showed that the protein level of Prx V remarkably decreased in a dose-dependent manner, whereas Prx II expression did not change. With the treatment prolongation of 4 μmol/L Cisplatin in A549 cells, Bcl-2 and caspase-3 were downregulated, while BAD upregulated.

CONCLUSIONS: Cisplatin treatment induces the ROS production, increases the apoptotic rate and downregulates the Prx expression in A549 cells.

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