title: Antiserum detection of reactive carbonyl species-modified DNA in human colonocytes.
abstract: Polyunsaturated fats have been linked to occurrences of sporadic colon cancer. One possible cause may be degradation of polyunsaturated fats during cooking, resulting in multiple reactive carbonyl species (RCS) that can damage nuclear DNA and proteins, particularly in rapidly dividing colon crypt cells. This study describes a novel antiserum against RCS-modified DNA, with apparent order of reactivity to DNA modified with 4-hydroxy-trans-2-nonenal > glyoxal > acrolein > crotonaldehyde > malondialdehyde; some reactivity was also observed against conjugated Schiff base-type structures. Anti-(RCS-DNA) antiserum was successfully utilised to demonstrate formation of RCS-DNA in a human colon cell model, exposed to RCS insult derived from endogenous and exogenous lipid peroxidation sources. Further utilisation of the antiserum for immunohistochemical analysis confirmed RCS-modified DNA in crypt areas of 'normal' colon tissue. These results fully support a potential role for dietary lipid peroxidation products in the development of sporadic colon cancer.
authors.affiliation: Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK
authors.forename: Nalini
authors.initials: N
authors.lastname: Mistry
authors.forename: Ruth J
authors.initials: RJ
authors.lastname: Bevan
authors.forename: Marcus S
authors.initials: MS
authors.lastname: Cooke
authors.forename: Mark D
authors.initials: MD
authors.lastname: Evans
authors.forename: Eugene P
authors.initials: EP
authors.lastname: Halligan
authors.forename: Damon A
authors.initials: DA
authors.lastname: Lowes
authors.forename: Karen
authors.initials: K
authors.lastname: Nichol
authors.forename: Joseph
authors.initials: J
authors.lastname: Lunec
mesh_terms: D000818:Animals; D002417:Cattle; D045744:Cell Line, Tumor; D003106:Colon / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D004247:DNA / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000378:metabolism; D004848:Epithelium / Q000473:pathology; D005224:Fats, Unsaturated / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D015227:Lipid Peroxidation; D011817:Rabbits; D012545:Schiff Bases / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism
chemical_list: D004273:DNA, Neoplasm; D005224:Fats, Unsaturated; D007074:Immunoglobulin G; D012545:Schiff Bases; D012709:Serum Albumin; D004247:DNA