title: The seleno-organic compound ebselen impairs mitochondrial physiology and induces cell death in AR42J cells.
abstract: Ebselen is a seleno-organic compound that causes cell death in several cancer cell types. The mechanisms underlying its deleterious effects have not been fully elucidated. In this study, the effects of ebselen (1 μM-40 μM) on AR42J tumor cells have been examined. Cell viability was studied using AlamarBlue(®) test. Cell cycle phase determination was carried out by flow cytometry. Changes in intracellular free Ca(2+) concentration were followed by fluorimetry analysis of fura-2-loaded cells. Distribution of mitochondria, mitochondrial Ca(2+) concentration and mitochondrial membrane potential were monitored by confocal microscopy of cells loaded with Mitotracker Green™ FM, rhod-2 or TMRM respectively. Caspase-3 activity was calculated following the luorogenic substrate ACDEVD-AMC signal with a spectrofluorimeter. Results show that cell viability decreased in the presence of ebselen. An increase in the number of cells in the S-phase of the cell cycle was observed. Ebselen induced a concentration-dependent mobilization of Ca(2+) from agonist- and thapsigargin-sensitive Ca(2+) pools. Ebselen induced also a transient increase in mitochondrial Ca(2+) concentration, a progressive decrease of the mitochondrial membrane potential and a disruption of the mitochondrial network. Finally, a concentration-dependent increase in caspase-3 activity was detected. We conclude that ebselen exerts deleterious actions on the cells that involve the impairment of mitochondrial physiology and the activation of caspase-3-mediated apoptotic pathway.
authors: Cell Physiology Research Group, Department of Physiology, University of Extremadura, Avenida Universidad s/n, Caceres E-10003, Spain.
authors: Patricia
authors: P
authors: Santofimia-Castaño
authors: Cell Physiology Research Group, Department of Physiology, University of Extremadura, Avenida Universidad s/n, Caceres E-10003, Spain.
authors: Lourdes
authors: L
authors: Garcia-Sanchez
authors: Facultade de Agronomia & Medicina Veterinaria, Universidade de Brasilia, 70900-100, Brasilia DF, Brazil.
authors: Deborah Clea
authors: DC
authors: Ruy
authors: Department of Gastroenterology, San Pedro de Alcantara Hospital, E-10003, Caceres, Spain.
authors: Miguel
authors: M
authors: Fernandez-Bermejo
authors: Cell Physiology Research Group, Department of Physiology, University of Extremadura, Avenida Universidad s/n, Caceres E-10003, Spain.
authors: Gines M
authors: GM
authors: Salido
authors: Cell Physiology Research Group, Department of Physiology, University of Extremadura, Avenida Universidad s/n, Caceres E-10003, Spain. Electronic address: agmateos@unex.es.
authors: Antonio
authors: A
authors: Gonzalez
mesh_terms: D000818:Animals; D001393:Azoles / Q000633:toxicity*; D002118:Calcium / Q000032:analysis; D053148:Caspase 3 / Q000187:drug effects / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry; D054833:Isoindoles; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000737:chemistry / Q000187:drug effects*; D016566:Organoselenium Compounds / Q000633:toxicity*; D010190:Pancreatic Neoplasms / Q000378:metabolism; D051381:Rats
chemical_list: D001393:Azoles; D054833:Isoindoles; D016566:Organoselenium Compounds; C042986:ebselen; D053148:Caspase 3; D002118:Calcium
keywords: Calcium; Cancer; Cell cycle; Ebselen; Proliferation; Red-ox state