title: Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)-phthalate (MEHP) on MA-10 Leydig cells and protection by selenium.
abstract: Di(2-ethylhexyl)-phthalate (DEHP) is the most abundantly used phthalate derivative, inevitable environmental exposure of which is suspected to contribute to the increasing incidence of testicular dysgenesis syndrome in humans. Oxidative stress and mitochondrial dysfunction in germ cells are suggested to contribute to phthalate-induced disruption of spermatogenesis in rodents, and Leydig cells are one of the main targets of phthalates' testicular toxicity. Selenium is known to be involved in the modulation of intracellular redox equilibrium, and plays a critical role in testis, sperm, and reproduction. This study was aimed to investigate the oxidative stress potential of DEHP and its consequences in testicular cells, and examine the possible protective effects of selenium using the MA-10 mouse Leydig tumor cell line as a model. In the presence and absence of selenium compounds [30 nM sodium selenite (SS), and 10 μM selenomethionine (SM)], the effects of exposure to DEHP and its main metabolite mono(2-ethylhexyl)-phthalate (MEHP) on the cell viability, enzymatic and non-enzymatic antioxidant status, ROS production, p53 expression, and DNA damage by alkaline Comet assay were investigated. The overall results of this study demonstrated the cytotoxicity and genotoxicity potential of DEHP, where MEHP was found to be more potent than the parent compound. SS and SM produced almost the same level of protection against antioxidant status modifying effects, ROS and p53 inducing potentials, and DNA damaging effects of the two phthalate derivatives. It was thus shown that DEHP produced oxidative stress in MA-10 cells, and selenium supplementation appeared to be an effective redox regulator in the experimental conditions used in this study, emphasizing the critical importance of the appropriate selenium status.
authors.affiliation: CEA Grenoble, INAC/SCIB/LAN, 17 Rue des Martyrs, 38054 Grenoble Cedex 9, France.
authors.forename: Pinar
authors.initials: P
authors.lastname: Erkekoglu
authors.forename: Walid
authors.initials: W
authors.lastname: Rachidi
authors.forename: Ozge Gursoy
authors.initials: OG
authors.lastname: Yuzugullu
authors.forename: Belma
authors.initials: B
authors.lastname: Giray
authors.forename: Alain
authors.initials: A
authors.lastname: Favier
authors.forename: Mehmet
authors.initials: M
authors.lastname: Ozturk
authors.forename: Filiz
authors.initials: F
authors.lastname: Hincal
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D045744:Cell Line, Tumor; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects; D004051:Diethylhexyl Phthalate / Q000031:analogs & derivatives* / Q000633:toxicity; D007985:Leydig Cells / Q000187:drug effects / Q000473:pathology; D008297:Male; D051379:Mice; D009152:Mutagenicity Tests; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012645:Selenomethionine / Q000494:pharmacology*; D018038:Sodium Selenite / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism
chemical_list: D000975:Antioxidants; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D012645:Selenomethionine; D004051:Diethylhexyl Phthalate; C016599:mono-(2-ethylhexyl)phthalate; D018038:Sodium Selenite