Osimertinib induces autophagy and apoptosis via reactive oxygen species generation in non-small cell lung cancer cells

• Published in: Toxicology and applied pharmacology 2017-04, Vol.321, p.18-26
• Main Authors: Tang, Zheng-Hai, Cao, Wen-Xiang, Su, Min-Xia, Chen, Xiuping, Lu, Jin-Jian
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; A549 Cells; Acrylamides; Aniline Compounds; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; APOPTOSIS; Apoptosis - drug effects; Apoptosis - physiology; Autophagy; Autophagy - drug effects; Autophagy - physiology; AZD9291; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - metabolism; CATALASE; Cell Survival - drug effects; Cell Survival - physiology; CYSTEINE; Dose-Response Relationship, Drug; FLUORESCENCE; GROWTH FACTORS; HCT116 Cells; Humans; Lung Neoplasms - drug therapy; Lung Neoplasms - metabolism; LUNGS; MICROTUBULES; MUTATIONS; NEOPLASMS; Osimertinib; PATIENTS; PHOSPHOTRANSFERASES; Piperazines - pharmacology; Piperazines - therapeutic use; PLASMIDS; Reactive Oxygen Species - metabolism; RECEPTORS; ROS; TYROSINE; VITAMIN E
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2017.02.017

Osimertinib (OSI), also known as AZD9291, is a third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor that has been approved for the treatment of non-small cell lung cancer (NSCLC) patients harboring EGFR T790M mutation. Herein, we indicated for the first time that OSI increased the accumulations of cytoplasmic vacuoles, the expression of phosphatidylethanolamine-modified microtubule-associated protein light-chain 3 (LC3-II), and the formation of GFP-LC3 puncta in various cancer cells. The OSI-induced expression of LC3-II was further increased when combined treatment with chloroquine (CQ), an autophagy inhibitor, and the mRFP-EGFP-LC3 plasmid-transfected cells exposed to OSI led to the production of more red-fluorescent puncta than green-fluorescent puncta, indicating OSI induced autophagic flux in the NSCLC cells. Knockdown of EGFR showed no effect on the OSI-induced expression of LC3-II in NCI-H1975 cells. In addition, OSI increased reactive oxygen species (ROS) generation and scavenge of ROS via pretreatment with N-acetyl-l-cysteine (NAC), catalase (CAT), or vitamin E (Vita E) significantly inhibited OSI-induced the accumulations of cytoplasmic vacuoles, the expression of LC3-II, as well as the formation of GFP-LC3 puncta. Combinative treatment with CQ could not remarkably change the OSI-induced cell viability decrease, whereas the OSI-induced cell viability decrease and apoptosis could be reversed through pretreatment with NAC, CAT, and Vita E, respectively. Taken together, this is the first report that OSI induces an accompanied autophagy and the generation of ROS is critical for the OSI-induced autophagy, cell viability decrease, and apoptosis in NSCLC cells. •Osimertinib induced the expressions of cytoplasmic vacuoles and autophagic markers in different cancer cells.•Osimertinib induced autophagic flux in NSCLC NCI-H1975 and HCC827 cell lines.•ROS generation contributed to osimertinib-induced cytoplasmic vacuoles, autophagy, cell viability decrease, and apoptosis.