title: Reactive oxygen species involvement in apoptosis and mitochondrial damage in Caco-2 cells induced by enniatins A, A₁, B and B₁.
abstract: The cytotoxic effects, the generation of reactive oxygen species (ROS) and lipid peroxidation (LPO) as well as the cell cycle disruption, the induction of apoptosis and changes in mitochondrial membrane potential (ΔΨm) as a function of increasing time have been determined in human colorectal adenocarcinoma (Caco-2) cells after exposure to enniatins (ENs) A, A₁, B and B₁. IC₅₀ values obtained by the MTT and Neutral Red assay, after 24, 48 and 72 h of exposure ranged from 0.5±0.1 to >15 μM. A significant increase (p≤0.05) in ROS generation and LPO production, as determined by the fluorescent probe H2-DCFDA and TBARS method respectively, was observed for all mycotoxins tested at 3.0 μM concentration. The highest increase in ROS generation (2.6 fold higher than control) and LPO production (111%, as compared to control) was observed with EN A. Cell cycle was significantly arrested at G2/M phase after 24 h of exposure to EN A, A₁, B₁, whereas after 72 h of exposure an arrest in S phase was observed almost for all mycotoxins tested. Moreover, after 24 and 48 h of exposure, ENs increased the early apoptotic cells, whereas after 72h of exposure necrosis was observed. In addition the loss of ΔΨm was produced on Caco-2 cells after ENs exposure. ENs A, A₁, B and B₁ cytotoxicity involved early ROS generation that induced LPO oxidative damage, apoptosis and necrosis via the mitochondrial pathway. ENs A, A₁ and B₁ induced DNA damage. However the same effects cannot be proposed for EN B. Further studies on the toxicological effects induced by ENs A, A₁, B and B₁ are needed.
authors.affiliation: Laboratori de Toxicologia, Facultat de Farmacia, Universitat de Valencia, Av. Vincent Andres Estelles, 46100 Burjassot, Valencia, Spain. alessandra.prosperini@uv.es
authors.forename: A
authors.initials: A
authors.lastname: Prosperini
authors.forename: A
authors.initials: A
authors.lastname: Juan-García
authors.forename: G
authors.initials: G
authors.lastname: Font
authors.forename: M J
authors.initials: MJ
authors.lastname: Ruiz
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D018938:Caco-2 Cells; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D020552:Comet Assay; D047630:Depsipeptides / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D007202:Indicators and Reagents; D015227:Lipid Peroxidation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects*; D009942:Organometallic Compounds; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D004396:Coloring Agents; D047630:Depsipeptides; D007202:Indicators and Reagents; D009942:Organometallic Compounds; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; D013844:Thiazoles; C100264:enniatins; C110932:tetramethyl rhodamine ethyl ester; C022616:thiazolyl blue
keywords: Apoptosis; Caco-2 cells; Cell cycle; Cytotoxicity; DNA damage; Enniatins; Mitochondrial membrane potential; Oxidative stress