Cisplatin Activates the Growth Inhibitory Signaling Pathways by Enhancing the Production of Reactive Oxygen Species in Non-small Cell Lung Cancer Carrying an EGFR Exon 19 Deletion.

Abstract

Cisplatin is a potent anticancer drug for treating several types of cancer, including non-small-cell lung cancer (NSCLC). In this study, we investigated the cytotoxicity and mechanism of action of cisplatin in the human NSCLC cell line PC9. PC9 cells were treated with cisplatin for 72 h and then evaluated by a cell viability assay, DAPI staining, Giemsa staining, apoptosis assay, membrane permeability assay, cell cycle assay, ROS assay, SA-β-gal staining, TUNEL assay and Western blotting. Our findings revealed that the cytotoxic activity was associated with an apoptotic signaling pathway in response to DNA damage. Cisplatin exerted a significant concentration-dependent antiproliferative effect on PC9 cells. Cells subjected to cisplatin treatment showed morphological indications of apoptosis. Cell cycle arrest was related to the restriction of E2F-1 action by the cyclin-dependent protein kinase inhibitor p21WAF1/CIP1 Cisplatin induced apoptosis of PC9 cells by upregulating Fas, FasL, Bak, and tBID expression and PARP proteolytic cleavage. Cisplatin also reduced the mitochondrial membrane potential (MMP) and initiated a caspase cascade. Furthermore, the apoptotic impact of cisplatin depended on reactive oxygen species (ROS), as confirmed by ROS generation. Cisplatin induced anticancer effects through cell cycle arrest, ROS generation and caspase activation, resulting in cell apoptosis. Overall, the results show the mechanism by which cisplatin works as an anticancer drug in the treatment of NSCLC.