Erlotinib induces the human non–small‐cell lung cancer cells apoptosis via activating ROS‐dependent JNK pathways

Abstract

Although erlotinib (ERL) has drawn more and more attention toward its anticancer properties effect, the underlying mechanisms of ERL's anticancer properties effect remain unclear yet. So, the aim of this research was to explore the underlying anticancer mechanisms of ERL and to explore whether the reactive oxygen species (ROS)‐dependent c‐Jun N‐terminal kinase (JNK) pathway contributed to the anticancer properties provided by ERL. In our study, we used MTT assay to detect the anticell growth ability of ERL on human non–small‐cell lung cancer cell lines (A549). The extent of cell apoptosis was determined by Hoechst 33342 staining and fluorescence‐activated cell sorter (FACS) assay. Then, DCFH‐DA and JC‐1 staining were used to monitor intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (MMP), respectively. Finally, the effect of ERL on phosphorylation state of JNK protein and downstream apoptosis concerned proteins were detected by western blotting assay. Results showed that ERL significantly suppressed the growth and reproduction of A549 cells with the concentration rising up in vitro. Hoechst 33342 staining and FACS assay also confirmed the proapoptosis effect of ERL on A549 cells with the concentration rising up. Furthermore, exposure of A549 cells to ERL increased the intracellular ROS production. As expected, intracellular ROS activated the proapoptotic JNK signaling pathway and inhibited the activation of EFGR signaling pathway. Our results also revealed that ERL could induce cell‐cycle arrest at G0/G1 period. Activation of JNK protein decreased MMP and downregulated content of antiapoptotic protein Bcl‐2 concomitant with the upregulated content of proapoptotic protein Bax in A549 cells. In addition, c‐Jun and cleaved caspase‐3 were also activated by the phosphorylated JNK induced by ERL. All of these proapoptosis effect of ERL was reversed by administration of N‐acetylcysteine (NAC), which performed as a ROS scavenger. Our results suggest that ERL induces A549 cells apoptosis via activating ROS‐dependent JNK pathways in human non–small lung cancer cells that provide a new experimental foundation for cancer therapy.

RETRACTED ARTICLE: Excessive mitochondrial fragmentation triggered by erlotinib promotes pancreatic cancer PANC-1 cell apoptosis via activating the mROS-HtrA2/Omi pathways

“…Our study proposed that the anti-cancer property of erlotinib relied on the activation of mitochondrial fragmentation by upregulating mitochondrial fission and downregulating mitochondrial fusion. Notably, the dose selection of ERL was according to a previous study [ 26 ] and this selection may be also relied on the types of cancer cell lines. In clinical practice, different doses of ERL have been used according to the tumor staging and pathologic grading.…”

“…No. SML2156) were incubated with the cancer cells for 24 h, and these concentrations of ERL were chosen according to a previous study [ 26 ]. FCCP (5 μm, Selleck Chemicals, Houston, TX, USA) and mitochondrial division inhibitor 1 (Mdivi1; 10 mM; Sigma-Aldrich; Merck KGaA) were used to activate and inhibit mitochondrial fragmentation, respectively, according to a previous study.…”

ROS in cancer therapy: the bright side of the moon

“…The induction of apoptosis by elevated ROS levels has been highlighted as the central mechanism responsible for the positive effects of monoclonal antibodies 85 and tyrosine kinase inhibitors 86 , which represent the core of targeted cancer therapy 87 . Among tyrosine kinase inhibitors, imatinib (a PDGFR inhibitor) and erlotinib (an EGFR inhibitor) induce ROS-dependent apoptosis in melanoma and non-small-cell lung cancer cells, respectively, through disruption of mitochondrial membrane potential upon the stimulation of JNK and p38 phosphorylation 88,89 , while vemurafenib (a BRAF inhibitor) increases the production of superoxide anions with the commensurate depolarization of the mitochondrial membranes in melanoma cells 90 . Among the monoclonal antibodies, rituximab (specific to the calcium-channel protein CD20 on the Fig.…”

Metabolic Remodelling: An Accomplice for New Therapeutic Strategies to Fight Lung Cancer

“…In A549 NSCLC cell line, erlotinib drove ROS-mediated apoptosis via activation of the c-Jun N-terminal kinase (JNK) pathway, leading ultimately to EGFR inhibition [183]. Administration of the ROS scavenger N-acetyl cysteine reversed this phenomenon [184].…”