Lysosomes as a possible target of enniatin B-induced toxicity in Caco-2 cells.
Enniatins are cyclic hexadepsipeptidic mycotoxins with ionophoric, antibiotic, and insecticidal activity. Enniatin B (EnnB), the most important analogue, is produced by many Fusarium species and is a common contaminant in grain-based foods. The compound's cytotoxic potential has been shown in different experiments; however, the mode of action has not been detailed so far. In the present study, several mutually confirmative experiments have been performed indicating that EnnB-initiated cytotoxicity could be connected with lysosomal membrane permeabilization (LMP). Lysosomal functionality, as assessed by the Neutral Red assay, was already affected after 3 h of toxin exposure. After 24 h, cell proliferation was decreased, and there was indication for a cell cycle arrest in the G(2)/M phase leading to the initiation of apoptosis or necrosis. Intracellular ROS-production was observed. However, antioxidants did not alter the observed EnnB-induced loss of lysosomal functionality leading to the conclusion that ROS was not an initial factor but one produced later in the event cascade. The collected data suggested that lysosomal destabilization is an upstream event in EnnB-initiated cytotoxicity followed by a certain extent of translocation of cathepsins into the cytosol, which was observed using immunological and proteomic methods. It appeared that cell death induced by EnnB was delayed and occurred not as a massive lysosomal breakdown but was probably progressing and leading to partial and selective LMP, starting a nonapoptotic cell death pathway with morphological features that had been previously considered as necrotic. The molecular mechanism of EnnB-triggered lysosomal destabilization, and the cellular processes leading to mitochondrial permeabilization and cell death are still unknown. They may, however, be connected to the compound's ionophoric properties.
Norwegian Veterinary Institute, Oslo, Norway. lada.ivanova@vetinst.no
L
L
Ivanova
W M
WM
Egge-Jacobsen
A
A
Solhaug
E
E
Thoen
C K
CK
Fæste
D017209:Apoptosis / Q000187:drug effects; D018938:Caco-2 Cells; D002403:Cathepsins / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D047630:Depsipeptides / Q000633:toxicity*; D005670:Fusarium / Q000378:metabolism; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism
D047630:Depsipeptides; D017382:Reactive Oxygen Species; C100264:enniatins; D002403:Cathepsins