title: Cytotoxicity and DNA-protein crosslink formation in rat nasal tissues exposed to vinyl acetate are carboxylesterase-mediated.
abstract: Vinyl acetate is used in the paint, adhesive, and paper board industries. Vinyl acetate is a nasal carcinogen in rats exposed by inhalation for 2 years to 200 and 600 ppm, but not 50 ppm. Previous studies from our laboratory suggest that rat liver microsome-activated vinyl acetate induces plasmid DNA-histone crosslinks, in vitro, through esterase-mediated metabolism. Since nasal tissues contain high levels of carboxylesterase, tumorigenesis may be related to in situ production of the hydrolysis products acetaldehyde and acetic acid. Vinyl acetate was cytotoxic to both respiratory and olfactory tissues in vitro at 50-200 mM, but not 25 mM, after 2 hr exposure. Pretreatment of rats with the carboxylesterase inhibitor, bis-(p-nitrophenyl) phosphate (BNPP), attenuated the cytotoxic effects and metabolism of vinyl acetate in both tissue types. Semicarbazide, an aldehyde scavenger, was unable to protect the tissues from vinyl acetate-induced cytotoxicity. When the metabolites were tested, acetic acid, but not acetaldehyde, was cytotoxic to both tissues. The induction of DNA-protein crosslink (DPXL) formation by acetaldehyde and vinyl acetate in rat nasal epithelial tissues was detected using a sodium dodecyl sulfate/KCl precipitation technique. Endogenous crosslink levels ranged from 0.5 to 2.0% of total DNA and were considered background. Epithelial cells isolated from both olfactory and respiratory turbinates exhibited dose- and time-dependent increases in DPXL formation when exposed to 10-150 mM acetaldehyde for 1-2 hr at 37 degrees C. Similarly, respiratory and olfactory epithelial cells exposed to 5-75 mM vinyl acetate for 1-2 hr accumulated up to 12- and 15-fold higher crosslink levels than untreated cells, respectively. However, vinyl acetate appears to induce much higher levels of DPXLs at equimolar doses than acetaldehyde. This is thought to be related to stimulation of acetaldehyde-induced DPXL formation by the pH lowering effect of acetic acid production (via vinyl acetate hydrolysis). Pretreatment of the nasal turbinates with 1 mM BNPP reduced 25 mM vinyl acetate-induced DPXL formation by over 75% in both tissues. These data support a hypothesis that carboxylesterase-mediated hydrolysis of vinyl acetate is necessary to generate the active intracellular cross-linking agent, acetaldehyde, and the cytotoxic metabolite, acetic acid.
authors.affiliation: Haskell Laboratory for Toxicology and Industrial Medicine, E. I. du Pont de Nemours and Co., Inc., Newark, Delaware 19714.
authors.forename: J R
authors.initials: JR
authors.lastname: Kuykendall
authors.forename: M L
authors.initials: ML
authors.lastname: Taylor
authors.forename: M S
authors.initials: MS
authors.lastname: Bogdanffy
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism / Q000633:toxicity; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D008297:Male; D009297:Nasal Mucosa / Q000378:metabolism; D009596:Nitrophenols / Q000494:pharmacology; D009666:Nose / Q000187:drug effects* / Q000473:pathology; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D012663:Semicarbazides / Q000494:pharmacology; D014420:Turbinates / Q000187:drug effects; D014753:Vinyl Compounds / Q000378:metabolism / Q000633:toxicity*
chemical_list: D009596:Nitrophenols; D011506:Proteins; D012663:Semicarbazides; D014753:Vinyl Compounds; C010059:carbamylhydrazine; C002887:bis(4-nitrophenyl)phosphate; D004247:DNA; D002265:Carboxylic Ester Hydrolases; D000079:Acetaldehyde; C011566:vinyl acetate