title: Cantharidin induces apoptosis of H460 human lung cancer cells through mitochondria-dependent pathways.
abstract: Lung cancer is one of the leading causes of death in cancer-related diseases. Cantharidin (CTD) is one of the components of natural mylabris (Mylabris phalerata Pallas). Numerous studies have shown that CTD induced cytotoxic effects on cancer cells. However, there is no report to demonstrate that CTD induced apoptosis in human lung cancer cells. Herein, we investigated the effect of CTD on the cell death via the induction of apoptosis in H460 human lung cancer cells. Flow cytometry assay was used for examining the percentage of cell viability, sub-G1 phase of the cell cycle, reactive oxygen species (ROS) and Ca²⁺ productions and the levels of mitochondrial membrane potential (∆Ψm). Annexin V/PI staining and DNA gel electrophoresis were also used for examining cell apoptosis. Western blot analysis was used to examine the changes of apoptosis associated protein expression and confocal microscopy for examining the translocation apoptosis associated protein. Results indicated that CTD significantly induced cell morphological changes and decreased the percentage of viable H460 cells. CTD induced apoptosis based on the occurrence of sub-G1 phase and DNA fragmentation. We found that CTD increased gene expression (mRNA) of caspase-3 and -8. Moreover, CTD increased ROS and Ca2+ production and decreased the levels of ∆Ψm. Western blot analysis results showed that CTD increased the expression of cleavage caspase-3 and -8, cytochrome c, Bax and AIF but inhibited the levels of Bcl-xL. CTD promoted ER stress associated protein expression such as GRP78, IRE1α, IRE1β, ATF6α and caspase-4 and it also promoted the expression of calpain 2 and XBP-1, but inhibited calpain 1 that is associated with apoptosis pathways. Based on those observations, we suggest that CTD may be used as a novel anticancer agent for the treatment of lung cancer in the future.
authors.affiliation: Graduate Institute of Chinese Medicine, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Te-Chun
authors.initials: TC
authors.lastname: Hsia
authors.affiliation: School of Pharmacy, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Yu
authors.affiliation: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Shu-Chun
authors.initials: SC
authors.lastname: Hsu
authors.affiliation: Graduate Institute of Chinese Medicine, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Nou-Ying
authors.initials: NY
authors.lastname: Tang
authors.affiliation: Department of Clinical Pathology, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C.
authors.forename: Hsu-Feng
authors.initials: HF
authors.lastname: Lu
authors.affiliation: Department of Physiology, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Huang
authors.affiliation: Division of Critical Care Medicine, Department of Medicine, Changhua Christian Hospital, Changhua, Taiwan, R.O.C.
authors.forename: Shin-Hwar
authors.initials: SH
authors.lastname: Wu
authors.affiliation: Graduate Institute of Chinese Medicine, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Jaung-Geng
authors.initials: JG
authors.lastname: Lin
authors.affiliation: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Jing-Gung
authors.initials: JG
authors.lastname: Chung
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D002193:Cantharidin / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D000091342:Endoplasmic Reticulum Chaperone BiP; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D017382:Reactive Oxygen Species; D053148:Caspase 3; D053181:Caspase 8; D002193:Cantharidin; D002118:Calcium