title: Cantharidin induces apoptosis in human bladder cancer TSGH 8301 cells through mitochondria-dependent signal pathways.
abstract: Cantharidin has shown potent anticancer activities on many types of human cancer cells. This study was performed to elucidate whether mitochondria and caspases are involved in the modulation of apoptosis and cell cycle arrest by cantharidin in human bladder cancer cells. The effect of cantharidin on cell cycle arrest, apoptosis, caspases, reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨ(m)) were measured by flow cytometry, and the levels of apoptosis-associated proteins and its regulatory molecules were studied by Western blotting. Cantharidin-induced apoptosis and DNA damage was determined by flow cytometric analysis, DAPI staining and Comet assay. After cantharidin treatment, the active forms of caspase-3, -8 and -9 were promoted. Cantharidin-induced apoptosis was associated with enhanced ROS and Ca(2+) generations, caused DNA damage, decreased the levels of ΔΨ(m) and promoted Endo G and AIF released from mitochondria. Cantharidin-induced G0/G1 arrest was associated with a marked decrease in the protein expressions of cyclin E and Cdc25c but promoted the levels of p21 and p-p53. Cantharidin-induced apoptosis was accompanied with up-regulation of the protein expression of Bax and PARP, but down-regulation of the protein levels of Bcl-2, resulting in dysfunction of mitochondria then led to Endo G and AIF release for causing induction of apoptosis.
authors.affiliation: Special Class of Healthcare, Industry Management, Central Taiwan University of Science and Technology, Taichung, Taiwan, ROC.
authors.forename: Jehn-Hwa
authors.initials: JH
authors.lastname: Kuo
authors.forename: Yung-Lin
authors.initials: YL
authors.lastname: Chu
authors.forename: Jai-Sing
authors.initials: JS
authors.lastname: Yang
authors.forename: Jing-Pin
authors.initials: JP
authors.lastname: Lin
authors.forename: Kuang-Chi
authors.initials: KC
authors.lastname: Lai
authors.forename: Hsiu-Maan
authors.initials: HM
authors.lastname: Kuo
authors.forename: Te-Chun
authors.initials: TC
authors.lastname: Hsia
authors.forename: Jing-Gung
authors.initials: JG
authors.lastname: Chung
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002193:Cantharidin / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002469:Cell Separation; D020552:Comet Assay; D005434:Flow Cytometry; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology*
chemical_list: D000970:Antineoplastic Agents; D017382:Reactive Oxygen Species; D002193:Cantharidin