21Background: Calotropis gigantea (CG) plant grows in Asia and tropical Africa. However, the precise mechanisms 22 of its anticancer effects have not yet been examined in human non-small cell lung cancer (NSCLC) cells, A54923 and NCI-H1299 cells.
24Purpose: This study was focused on the anti-cancer effects of CG extract on non-small cell lung cancer (NSCLC) 25 cells.
26Methods: The cytotoxic effects of CG extract on NSCLC, A549 and NCI-H1299 cells, were detected by MTS 27 assay, microscope and DAPI staining. Apoptosis was determined by annexin V-FITC/PI staining, cell cycle 28 analysis, western blotting, quantitative polymerase chain reaction, and JC-1 staining.
29Results: First, CG showed significant dose-dependent cytotoxicity in NSCLC, A549, and NCI-H1299 cells. In 30 addition to induction of caspase-8 processing, CG induced apoptosis by upregulating mRNA expression levels of 31 extrinsic pathway molecules such as Fas, Fas ligand (FasL), Fas-associated protein with death domain (FADD) 32and death receptor 5 (DR5). Also, mitochondrial membrane potential (MMP) was collapsed, and intrinsic pathway 33 molecules such as poly (ADP-ribose) polymerase (PARP), caspase-3, and caspase-9 were processed by CG.
34Moreover, reactive oxygen species (ROS) were generated in a CG dose-dependent manner, and inhibition of ROS 35 by NAC, ROS scavenger, recovered A549 and NCI-H1299 cell viability.
36Conclusion: These results indicate that CG causes apoptosis by activating the extrinsic and intrinsic pathways and 37 generating ROS in NSCLC cells. These results suggest that CG can be used as a lung cancer therapeutic agent. 38 39 Keyword: Apoptosis, non-small cell lung cancer, Calotropis gigantea, anti-cancer, intrinsic and extrinsic 40 pathways, ROS 41 42 Abbreviations used: CG, Calotropis gigantea; NSCLC, non-small cell lung cancer; FasL, Fas Ligand; 43 FADD, Fas-associated protein with death domain; DR5, death receptor 5; PARP, poly (ADP-ribose) 44 polymerase; ROS, reactive oxygen species -3-