Induction of DNA damage by risk factors of colon cancer in human colon cells derived from biopsies.
In order to increase the understanding of the factors responsible for causing human colon cancer, a technique was developed to detect genotoxic effects of chemicals in human colon cells. Risk factors suspected to be associated with the aetiology of human colon cancer were subsequently investigated: the method is based on the measurement of DNA damage in primary cells freshly isolated from human colon biopsies with the single cell microgel ectrophoresis technique ('Comet Assay'). 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3-methyl-3H-imidazo[4,5f]quinoline (IQ), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), dinitrosocaffeidine (DNC) lithocholic acid (LCA), hydrogen peroxide (H2O2) and benzo[a]pyrene (B[a]P) were investigated for their genotoxic and cytotoxic effects following 30 min incubation with colon cells of human, and for comparative purposes also of the rat colon. The nitrosamides (MNNG, DNC) were very genotoxic in human colon cells. MNNG was more genotoxic in human than in rat colon cells. In contrast, the rat colon carcinogens PhIP and IQ were not genotoxic in human colon cells. PhIP did induce DNA damage in rat colon cells, which correlates to its capacity of inducing tumors in this animal tissue. LCA was toxic (rat > human) and concomitantly caused DNA damage in higher concentrations. The widespread contaminant B[a]P was not genotoxic in colon cells of either species using this system. H2O2 was found to be a potent genotoxic agent to both rat and human colon cells (human > rat). In summary, those compounds chosen as representatives of endogenously formed risk factors (MNNG, H2O2, LCA) have a higher toxic and/or genotoxic potency in human colon tissue than in rat colon. They are also more effective in this system than the contaminants tested so far (B[a]P, PhIP, IQ). The newly developed technique is rapid and yields relevant results. It is a novel and useful approach to assess different chemical compounds for genotoxic activities in tumour target tissues of the human.
Pool-Zobel
B L
BL
Institute for Nutritional Physiology, Federal Research Center for Nutrition, Karlsruhe, Germany.
Leucht
U
U
D000328:Adult; D000368:Aged; D000818:Animals; D001564:Benzo(a)pyrene / Q000494:pharmacology; D001706:Biopsy; D002478:Cells, Cultured; D003106:Colon; D004247:DNA / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007413:Intestinal Mucosa / Q000166:cytology*; D008095:Lithocholic Acid / Q000494:pharmacology; D008297:Male; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008875:Middle Aged; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000494:pharmacology*; D011804:Quinolines / Q000494:pharmacology; D051381:Rats; D012307:Risk Factors
D007093:Imidazoles; D009153:Mutagens; D011804:Quinolines; D008769:Methylnitronitrosoguanidine; D001564:Benzo(a)pyrene; D008095:Lithocholic Acid; D004247:DNA; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D006861:Hydrogen Peroxide; C036989:2-amino-3,4-dimethylimidazo(4,5-f)quinoline