title: Cantharidin induces G2/M phase arrest by inhibition of Cdc25c and Cyclin A and triggers apoptosis through reactive oxygen species and the mitochondria‑dependent pathways of A375.S2 human melanoma cells.
abstract: Cantharidin (CTD), a component of natural mylabris (Mylabris phalerata Pallas) was reported to have high cytotoxicity in many human cancer cell lines. However, it was not reported to affect human melanoma A375.S2 cells. In the present study, we found that CTD induced cell morphological changes and decreased the percentage of viable cells and induced G2/M phase arrest and induction of apoptosis in A375.S2 cells. Results also showed that CTD induced the generation of reactive oxygen species (ROS) and Ca2+ and decreased mitochondria membrane potential and lead to the release of cytochrome c, AIF and Endo G. Further investigation revealed that CTD induced A375.S2 cells with an increase of caspase activation and caspase-dependent apoptotic proteins to trigger correlated pathway mechanisms according to western blotting results. Western blotting was used for examining the changes of G2/M phase arrest and apoptosis-associated protein expression and confocal laser microscopy was used to examine the translocation apoptosis-associated protein. Results showed that CTD increased the protein expression of caspase-3, -8 and -9, cytochrome c, Bax, Bid, Endo G and AIF but inhibited the levels of Bcl-2 and Bcl-x. CTD induced ER stress-associated protein expression such as GRP78, IRE1β, ATF6α and caspase-12. Based on those observations, we suggest that CTD may have potential as a novel anti-cancer agent for the treatment of skin cancer.
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Yu-Ping
authors.initials: YP
authors.lastname: Hsiao
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Chung-Hung
authors.initials: CH
authors.lastname: Tsai
authors.affiliation: School of Pharmacy, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Ping-Ping
authors.initials: PP
authors.lastname: Wu
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: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
authors.forename: Hsin-Chung
authors.initials: HC
authors.lastname: Liu
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: Department of Dermatology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan, R.O.C.
authors.forename: Jen-Hung
authors.initials: JH
authors.lastname: Yang
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; D002193:Cantharidin / Q000008:administration & dosage*; D045744:Cell Line, Tumor; D019925:Cyclin A / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D000091342:Endoplasmic Reticulum Chaperone BiP; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008545:Melanoma / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors / Q000096:biosynthesis*
chemical_list: D019925:Cyclin A; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D009363:Neoplasm Proteins; D017382:Reactive Oxygen Species; C517009:CDC25C protein, human; D020687:cdc25 Phosphatases; D002193:Cantharidin