ABSTRACT
Fumonisin B1 (FB1), produced by the fungus Fusarium moniliforme, belongs to a class of sphingosine analogue mycotoxins that occur widely in the food chain. Epidemiological studies have associated consumption of Fusarium moniliforme-contaminated food with human oesophageal cancer in China and South Africa. FB1 also causes equine leucoencephalomalacia. Evidence for induction of apoptosis by FB1 was first obtained when C6 glioma cells were incubated with fumonisin B1 (3-27 microM) causing DNA fragmentation profiles showing DNA laddering in gel electrophoresis and apoptotic bodies revealed by chromatin staining with acridine orange and ethidium bromide. Further confirmation experiments and comet assays have been performed under similar conditions. The results of the comet test show that FB1 at 9 and 18 microM induces respectively 50 +/- 2% and 40 +/- 1% of cells with a comet with an increased tail length of 93 +/- 9 microm and 102 +/- 17 microm respectively. Under these concentrations, FB1 induced DNA fragmentation and laddering and many apoptotic bodies. Pre-incubation of the cells with vitamin E (25 microM) for 24 h before FB1 (18 microM) significantly reduced DNA fragmentation and apoptotic bodies induced by FB1.
Subject(s)
Carboxylic Acids/toxicity , Carcinogens, Environmental/toxicity , DNA Fragmentation/drug effects , Fumonisins , Vitamin E/pharmacology , Animals , Apoptosis/drug effects , Comet Assay , DNA Damage/drug effects , DNA, Neoplasm/drug effects , Electrophoresis, Agar Gel , Glioma , Image Processing, Computer-Assisted , Mutagens/toxicity , Rats , Tumor Cells, CulturedABSTRACT
Fumonisin B(1) produced by the fungus Fusarium moniliforme is a member of a new class of sphinganine analogue mycotoxins that occur widely in the food chain. Epidemiological studies associate FB(1) with human oesophageal cancer in China and South Africa. FB(1) also causes acute pulmonary edema in pigs and equine leucoencephalomalacia. This disease is thought to be a consequence of inhibition by FB(1) of cellular ceramide synthesis in cells. To investigate further on this pathogenesis, the effect of FB(1) was studied on cell viability (3 to 54 microM of FB(1)), protein (2.5 to 20 microM of FB(1)) and DNA syntheses (2.5 to 50 microM of FB(1)), and cellular cycle (3 to 18 microM of FB(1)) of rat C6 glioma cells after 24 h incubation. The results of the viability test show that FB(1) induces 10 +/- 2% and 47 +/- 4% cell death with, respectively, 3 and 54 microM, in C6 cells. This cytotoxicity induced by FB(1) was efficiently prevented when the cells were preincubated 24 h with vitamin E (25 microM). FB(1) displays epigenetic properties since it induced hypermethylation of the DNA (9-18 microM). Inhibition of protein synthesis was observed with FB(1) with an IC(50) of 6 microM showing that C6 glioma cells are very sensitive to FB(1); however, the synthesis of DNA was only slightly inhibited, up to 20 microM of FB1. The flow cytometry showed that the number of cells in phase S decreased significantly as compared to the control p = 0.01 from 18. 7 +/- 2.5% to 8.1 +/- 1.1% for 9 microM FB(1). The number of cells in phase G(2)/M increased significantly as compared to the control (p
Subject(s)
Carboxylic Acids/toxicity , DNA Methylation , DNA/drug effects , Fumonisins , Glioma/pathology , Mycotoxins/toxicity , Animals , Cell Cycle/drug effects , Cell Survival/drug effects , DNA/biosynthesis , Protein Biosynthesis , Rats , Tumor Cells, Cultured , Vitamin E/pharmacologyABSTRACT
In 3T3 mouse fibroblasts that were treated by surfactant toxins such as staphylococcal delta toxin, melittin from bee venom and lysolecithin, microsomal phospholipase A2 (PLA2) was activated. However, these toxins did not activate PLA2 when added to cell-free homogenates or microsomal preparations. The maximal activation was achieved at 1 mM Ca2+ pH 8.5. The microsomal PLA2 stimulated by these toxins had a high fatty acid specificity (C-2 position) toward arachidonic acid.
