Ferric iron is genotoxic in non-transformed and preneoplastic human colon cells.
Iron could be a relevant risk factor for carcinogenesis since it catalyses the formation of reactive oxygen species (ROS), which damage DNA. We previously demonstrated genotoxic effects by ferric iron using the human colon cancer cell line HT29. Here we investigated ferric iron in primary non-transformed colon cells and in a preneoplastic colon adenoma cell line (LT97), which both are suitable models to study effects of carcinogens during early stages of cell transformation. Genetic damage was determined using the Comet assay. Comet FISH (fluorescence in situ hybridization) was used to assess specific effects on TP53. Fe-NTA (0-1000 microM, 30 min, 37 degrees C) significantly induced single strand breaks in primary colon cells (500 microM Fe-NTA: Tail intensity [TI] 22.6%+/-5.0% versus RPMI control: TI 10.6%+/-3.9%, p<0.01) and in LT97 cells (1000 microM Fe-NTA: TI 26.8%+/-7.3% versus RPMI control: TI 11.1%+/-3.7%, p<0.01). With the Comet FISH protocol lower concentrations of Fe-NTA significantly increased DNA damage already at 100 and 250 microM Fe-NTA in primary colon and LT97 adenoma cells, respectively. This damage was detected as an enhanced migration of TP53 signals into the comet tail in both cell types, which indicates a high susceptibility of this tumor relevant gene towards Fe-NTA. In conclusion, Fe-NTA acts genotoxic in non-transformed and in preneoplastic human colon cells, in which it also enhances migration of TP53 at relatively low concentrations. Translated to the in vivo situation these results suggest that iron overload putatively contributes to a genotoxic risk during early stages of colorectal carcinogenesis on account of its genotoxic potential in non-tumorigenic human colon cells.
Knöbel
Y
Y
Institute of Nutrition, Department of Nutritional Toxicology, Friedrich-Schiller-University, Dornburger Str. 25, 07743 Jena, Germany.
Weise
A
A
Glei
M
M
Sendt
W
W
Claussen
U
U
Pool-Zobel
B L
BL
D000236:Adenoma / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D002869:Chromosome Aberrations; D003106:Colon / Q000187:drug effects*; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005290:Ferric Compounds / Q000633:toxicity*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D009566:Nitrates / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
D005290:Ferric Compounds; D009566:Nitrates; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; C025302:ferric nitrate