Accumulation of miscoding etheno-DNA adducts and highly expressed DNA repair during liver fluke-induced cholangiocarcinogenesis in hamsters.
Infection by Opisthorchis viverrini, a risk factor for cholangiocarcinoma (CCA) may act through chronic inflammation, oxidative stress and lipid peroxidation (LPO)-related damage and growth stimuli. 1,N6-etheno-2'-deoxyadenosine (epsilondA), and 3,N4-etheno-2'-deoxycytidine (epsilondC), markers for LPO-derived DNA damage were highly increased in white blood cell and urine of O. viverrini-infected Thai patients. In order to investigate tissue specificity etheno adducts were measured in a cholangiocarcinogenesis model, in O. viverrini-infected hamsters that had received N-nitrosodimethylamine (NDMA, 12.5 ppm in dw) for 2 months. epsilondA- and epsilondC-levels were analyzed in paraffin-embedded liver sections by a novel immunohistochemical method, from 21 up to 180 days post-O. viverrini-infection. In inflamed areas of the liver, etheno adducts were localized in the nuclei of inflammatory cells and in the epithelial lining of the bile duct. Semi-quantitative image analysis showed higher adduct levels in the liver of O. viverrini-infected hamsters, treated with or w/o NDMA when compared with untreated controls. Levels were found highest in the liver of O. viverrini-infected plus NDMA-treated hamsters. Adducts increased in an age-dependent manner from O. viverrini-infection until CCA development. Increased adduct formation paralleled histopathological changes in plasma alkaline phosphatase (ALP) activity, bile duct hyperplasia, dysplasia, precancerous lesions, and CCA appearance. Also elevated expression of alkyladenine DNA glycosylase (AAG), which excises 1,N6-ethenoadenine (epsilonA) was linked to higher adduct formation, suggesting imbalanced repair. Our results implicate accumulation of inflammation-related, promutagenic DNA damage in target tissue and possibly imbalanced repair in the onset of cholangiocarcinogenesis.
Dechakhamphu
Somkid
S
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Pinlaor
Somchai
S
Sitthithaworn
Paiboon
P
Bartsch
Helmut
H
Yongvanit
Puangrat
P
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001650:Bile Duct Neoplasms / Q000235:genetics* / Q000469:parasitology*; D001652:Bile Ducts / Q000378:metabolism; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000469:parasitology* / Q000473:pathology; D006224:Cricetinae; D018736:DNA Adducts / Q000378:metabolism*; D004260:DNA Repair; D003839:Deoxyadenosines / Q000378:metabolism*; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000378:metabolism; D005211:Fascioliasis / Q000150:complications*; D007249:Inflammation / Q000150:complications; D008099:Liver / Q000378:metabolism; D008297:Male
D018736:DNA Adducts; D003839:Deoxyadenosines; D003841:Deoxycytidine; C032221:1,N(6)-ethenodeoxyadenosine; C032222:3,N(4)-ethenodeoxycytidine; D000469:Alkaline Phosphatase