DNA-damaging potential and glutathione depletion of 2-cyclohexene-1-one in mammalian cells, compared to food relevant 2-alkenals.
2-Cyclohexene-1-one (CHX) occurs as a natural ingredient in some tropical fruits and has been detected as a contaminant in certain artificially sweetened soft drinks. To elucidate its cytotoxic/genotoxic effectiveness, CHX was tested in mammalian cell lines (V79 and Caco-2) and in primary human colon cells in comparison to structurally related 2-alkenals. Inhibition of cell growth (IC(50)) and cytotoxicity (LC(50)) were determined by protein staining with sulforhodamin B (SRB) and by trypan blue exclusion, respectively. DNA damage--both strand breaks and oxidised purines--was quantified by comet assay. Depletion of glutathione was measured in a kinetic assay, based on 5-thio-2-nitrobenzoate (TNB) formation. For CHX, a moderate cytotoxicity was observed after 1h incubation in V79 cells (LC(50): 4.75mM). The 2-alkenals ((E)-2-octenal (OCTE), (2E,4Z)-2,4-hexadienal (HEXDI), (E)-2-nonenal (NONE), (2E,6Z)-2,6-nonadienal (NONDI)) exhibited a distinctly higher cytotoxicity, except for (E)-2-hexenal (HEX) (LC(50): 3.67mM) and cinnamaldehyde (CA) (LC(50): 4.45mM). If the incubation time was prolonged to 24h, an IC(50) of 15microM was obtained for CHX which is well within the range obtained for the 2-alkenals (4 and 17microM). Concentration-dependent DNA damage was observed after 1h incubation with CHX. The respective DC(50) values (concentration inducing DNA damage in 50% of cells) were 272microM (V79) and 455microM (Caco-2). All 2-alkenals were more active under these conditions, except for CA. In primary human colon cells, CHX (800microM, 30min) exhibited a weak, but still significant DNA-damaging potential. Glutathione levels in V79 cells were effectively depleted (down to approximately 20%) by CHX concentrations not yet inducing DNA damage (c < or = 50microM). Incubation with CHX or 2-alkenals (50 and 100microM, 1h), followed by H2O2 treatment (5min, 25microM) resulted in increased levels of oxidised purines in the modified comet assay. CHX and HEX, additionally tested in primary human colon cells, depleted glutathione and increased the sensitivity towards oxidative stress.
Glaab
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University Department of Chemistry, Division of Food Chemistry and Environmental Toxicology, 67663, Kaiserslautern, Germany.
Collins
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Eisenbrand
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Janzowski
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D000447:Aldehydes / Q000633:toxicity; D000475:Alkenes / Q000633:toxicity*; D000818:Animals; D018938:Caco-2 Cells; D002460:Cell Line; D002478:Cells, Cultured; D006224:Cricetinae; D003512:Cyclohexanones / Q000633:toxicity*; D004249:DNA Damage; D005504:Food Analysis; D005506:Food Contamination; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D009153:Mutagens / Q000633:toxicity; D018384:Oxidative Stress; D011687:Purines / Q000737:chemistry / Q000378:metabolism; D013014:SOS Response, Genetics; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics
D000447:Aldehydes; D000475:Alkenes; D003512:Cyclohexanones; D009153:Mutagens; D011687:Purines; C028215:2-cyclohexen-1-one; D005978:Glutathione