title: Norcantharidin-induced Apoptosis of AGS Human Gastric Cancer Cells Through Reactive Oxygen Species Production, and Caspase- and Mitochondria-dependent Signaling Pathways.
abstract: Norcantharidin (NCTD) was purified from mylabris, the dried body of the Chinese blister beetle. NCTD has been shown to exhibit anticancer activities in many human cancer cell lines, but there are no reports to show whether it induces apoptosis of human gastric cancer cells. Therefore, in the present study, we investigated NCTD-induced cell death and associated protein expression in human gastric cancer AGS cells in vitro. Cell morphological changes, viability and cell-cycle distribution were examined and analyzed by phase-contrast microscopy and flow cytometric assays. Flow cytometry was also used to measure the levels of reactive oxygen species (ROS), Ca2+, mitochondrial membrane potential (Ψm) and activity of caspases. The results indicated that NCTD induced cell morphological changes, reduced total viable cell number and induced G0/G1phase arrest. NCTD also increased ROS production and reduced the Ψmand increased caspase-9 activity in AGS cells. Western blotting also found that NCTD increased the pro-apoptotic proteins such as BCL2-associated X protein (BAX) and BH3 interacting-domain death agonist (BID) and increased the release of cytochrome c, apoptosis inducing factor (AIF) and endonuclease G (Endo G) release from mitochondria in AGS cells. NCTD also significantly increased the expression of active forms of caspase-3 and -8 and -9 and reduced the expression of caspase-4 and -12 in AGS cells. Based on these observations, we suggest that NCTD-induced apoptotic cell death may be through mitochondria- and caspase-dependent pathways.
authors: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
authors: Li-Cheng
authors: LC
authors: Zheng
authors: Department of Surgery, China Medical University Hospital, Taichung, Taiwan, R.O.C.|Department of Clinical Nutrition, China Medical University Hospital, Taichung, Taiwan, R.O.C.|Terry Fox Cancer Research Laboratory, China Medical University Hospital, Taichung, Taiwan, R.O.C.
authors: Mei-Due
authors: MD
authors: Yang
authors: Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan, R.O.C.
authors: Chao-Lin
authors: CL
authors: Kuo
authors: Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan, R.O.C.
authors: Chia-Hsin
authors: CH
authors: Lin
authors: Department of Biotechnology, Asia University, Taichung, Taiwan, R.O.C.
authors: Ming-Jen
authors: MJ
authors: Fan
authors: Division of Neurosurgical Oncology, Neurological Institute, Taichung Veterans General Hospital, Taichung, Taiwan, R.O.C.
authors: Yu-Cheng
authors: YC
authors: Chou
authors: Department of Clinical Pathology, Cheng Hsin General Hospital, Taipei, Taiwan, R.O.C.
authors: Hsu-Feng
authors: HF
authors: Lu
authors: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C.
authors: Wen-Wen
authors: WW
authors: Huang
authors: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C. jgchung@mail.cmu.edu.tw sfpeng@mail.cmu.edu.tw.
authors: Shu-Fen
authors: SF
authors: Peng
authors: Department of Biological Science and Technology, China Medical University, Taichung, Taiwan, R.O.C. jgchung@mail.cmu.edu.tw sfpeng@mail.cmu.edu.tw.|Department of Biotechnology, Asia University, Taichung, Taiwan, R.O.C.
authors: Jing-Gung
authors: JG
authors: Chung
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology*; D020169:Caspases / Q000378:metabolism*; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D013274:Stomach Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D017382:Reactive Oxygen Species; C069741:norcantharidin; D020169:Caspases
keywords: AGS human gastric cancer cells; Norcantharidin; ROS; apoptosis; mitochondria; reactive oxygen species