Formation of Acrolein-derived 2′-Deoxyadenosine Adduct in an Iron-induced Carcinogenesis Model

Abstract

Acrolein is a representative carcinogenic aldehyde found ubiquitously in the environment and formed endogenously through oxidation reactions, such as lipid peroxidation and myeloperoxidase-catalyzed amino acid oxidation. It shows facile reactivity toward DNA to form an exocyclic DNA adduct. To verify the formation of acrolein-derived DNA adduct under oxidative stress in vivo, we raised a novel monoclonal antibody (mAb21) against the acrolein-modified DNA and found that the antibody most significantly recognized an acroleinmodified 2 -deoxyadenosine. On the basis of chemical and spectroscopic evidence, the major antigenic product of mAb21 was the 1,N 6 -propano-2 -deoxyadenosine adduct. The exposure of rat liver epithelial RL34 cells to acrolein resulted in a significant accumulation of the acrolein-2 -deoxyadenosine adduct in the nuclei. Formation of this adduct under oxidative stress in vivo was immunohistochemically examined in rats exposed to ferric nitrilotriacetate, a carcinogenic iron chelate that specifically induces oxidative stress in the kidneys of rodents. It was observed that the acrolein-2 -deoxyadenosine adduct was formed in the nuclei of the proximal tubular cells, the target cells of this carcinogenesis model. The same cells were stained with a monoclonal antibody 5F6 that recognizes an acrolein-lysine adduct, by which cytosolic accumulation of acrolein-modified proteins appeared. Similar results were also obtained from myeloperoxidase knockout mice exposed to the iron complex, suggesting that the myeloperoxidase-catalyzed oxidation system might not be essential for the generation of acrolein in this experimental animal carcinogenesis model. The data obtained in this study suggest that the formation of a carcinogenic aldehyde through lipid peroxidation may be causally involved in the pathophysiological effects associated with oxidative stress.Lipid peroxidation leads to the formation of a broad array of different products with diverse and powerful biological activities. Among them are a variety of different aldehydes (1). The primary products of lipid peroxidation, lipid hydroperoxides (2), can undergo carbon-carbon bond cleavage via alkoxyl radicals in the presence of transition metals, giving rise to the formation of short chain, unesterified aldehydes (2, 3) or a second class of aldehydes still esterified to the parent lipid. These reactive aldehydic intermediates readily form covalent adducts with cellular macromolecules, including DNA, leading to disruption of important cellular functions and mutations. The important agents that give rise to the modification of DNA may be represented by ␣,␤-unsaturated aldehydic intermediates, such as 2-alkenals and 4-hydroxy-2-alkenals (4, 5). 2-Alkenals represent a group of highly reactive aldehydes containing two electrophilic reaction centers. A partially positive carbon 1 or 3 in such molecules can attack nucleophiles, such as protein and DNA, which leads to the formation of cyclic adducts or cross-links. Among all of the ␣,␤-unsaturated aldehydes, acro...

Determination of Urine 3-HPMA, a Stable Acrolein Metabolite in a Rat Model of Spinal Cord Injury

“…Taken together, measurement of urine 3-HPMA may represent a novel theranostic approach for SCI. Since acrolein may potentially be linked to many other conditions, such as cancer, 2,[25][26][27][28] aging, 29 pollution, 30,31 and smoking, 32,33 the benefits of this study has the potential to significantly enhance human health.…”

Antiserum detection of reactive carbonyl species-modified DNA in human colonocytes

“…Antibodies, both poly-and mono-clonal, have been generated against RCS-modified DNA bases. For example, MDA-deoxyguanosine [28], deoxycytidine-glyoxal [29], deoxyadenosine-acrolein [30], 1,N(6)-ethenodeoxyadenosine [31] and ethenodeoxyguanosine [32]. These specific RCS-DNA antibodies have been successfully used to measure carbonyl stress in human and animal studies following oxidative and peroxidative insult [33,34].…”

1,N6-α-hydroxypropanoadenine, the acrolein adduct to adenine, is a substrate for AlkB dioxygenase

“…As the lesion is efficiently formed in vivo after exposure to acrolein (12), it is likely to be harmful to living organisms.…”

“…The only (to the best of our knowledge) work describing possible mutagenicity of HPA is reference (11) where 22% of base substitutions induced by ACR in human fibroblasts were found at A•T sites. The formation of this adduct was also shown in rat liver epithelial RL34 cells exposed to acrolein and in rats exposed to Fe 3+ NTA, a carcinogenic iron chelate that specifically induces oxidative stress in the kidney of rodents (12).…”