Bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE) are used as starting substances for the manufacturing of epoxy resins used in internal can coatings. They are obtained by a condensation reaction between epichlorohydrin with bisphenol A and bisphenol F, respectively. These potential endocrine disrupting chemicals are able to enter the food chain and to reach the intestinal epithelium, causing structural and functional damages. The human colorectal adenocarcinoma cell line Caco-2 is a widely used in vitro model of the intestinal cells. The aim of this study was to characterize BADGE and BFDGE toxicity in Caco-2 cells, in particular, at the cellular and molecular level. Using several approaches, we characterized BADGE- and BFDGE-induced cell toxicity in Caco-2 cells. The treatment was done using different concentrations up to cytotoxic doses and different times of exposure to the agents. We evaluated the effect of these compounds on cell morphology, cell detachment, cell proliferation, F-actin disruption and plasma membrane integrity. Both compounds are able to induce morphological changes, cell detachment from the substratum and to inhibit cell proliferation, being these effects time and dose-dependent. Moreover, BADGE and BFDGE induce F-actin depolymerization, this effect is very potent at 24 h of incubation with the agents and a complete F-actin disruption can be observed at 200 microM BADGE or BFDGE. In addition, cell integrity is not damaged, since neither propidium iodide uptake nor LDH release takes place in Caco-2 cells exposed to high doses of these agents for 24 h.

Palytoxin is a marine toxin responsible for a fatal type of poisoning in humans named clupeotoxism, with symptoms such as neurologic disturbances. It is believed that it binds to the Na+/K+‐ATPase from the extracellular side and modifies cytosolic ions; nevertheless, its effects on internal cell structures, such as the cytoskeleton, which might be affected by these initial events, have not been fully elucidated. Likewise, ostreocin‐D, an analog of palytoxin, has been only recently found, and its action on excitable cells is therefore unknown. Therefore, our aim was to investigate the modifications of ion fluxes associated with palytoxin and ostreocin‐D activities, and their effects on an essential cytoskeletal component, the actin system. We used human neuroblastoma cells and fluorescent dyes to detect changes in membrane potential, intracellular Ca2+ concentration, cell detachment, and actin filaments. Fluorescence values were obtained with spectrofluorymetry, laser‐scanning cytometry, and confocal microscopy; the last of these was also used for recording images. Palytoxin and ostreocin‐D modified membrane permeability as a first step, triggering depolarization and increasing Ca2+ influx. The substantial loss of filamentous actin, and the morphologic alterations elicited by both toxins, are possibly secondary to their action on ion channels. The decrease in polymerized actin seemed to be Ca2+‐independent; however, this ion could be related to actin cytoskeletal organization. Palytoxin and ostreocin‐D alter the ion fluxes, targeting pathways that involve the cytoskeletal dynamics of human excitable cells.

Palytoxin isolated from the genus Palythoa is the most potent marine toxin known. The aim of the present study was to quantify palytoxin-induced cellular injury in the human intestinal cell line Caco-2. Cellular damage was measured by evaluating cell proliferation, cell membrane permeability, cell morphology and apoptotic markers. Furthermore, changes in F-actin were studied after exposure of cells to increasing amounts of palytoxin. The results show that cell proliferation decreased in a concentration-dependent manner with a mean IC(50) value of about 0.1 nM. A noticeable increase of cell detachment correlated with cell rounding and F-actin depolymerization was observed in palytoxin-treated cells. Moreover LDH was released from the cells in a dose and time dependent manner, although under these conditions there was no propidium iodide uptake. On the other hand, palytoxin impaired mitochondrial activity but other apoptotic markers, such as DNA fragmentation or caspases activation, were not observed. The results obtained in this paper suggest that the effects of palytoxin in Caco-2 cells were very potent and unspecific, since a primary necrosis and a secondary apoptosis seem to occur under these conditions.

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