sodium-nitrite and Cell-Transformation--Neoplastic

Data from literature as well as the results of the author's investigations on the genotoxic and carcinogenic effects of sodium nitrite (SN) are presented. Carcinogenicity of SN detected in animal experiments proved to be connected with the formation of nitroso compounds from endogenous nitrosable precursors. Sodium nitrite possessed the transforming and promoting effects in the cell cultures as well as the mutagenic effects in the bacterial systems where predominant effect of SN as compared with N-nitrosodimethylamine (NDMA) was registered. The prolonged pretreatment with SN amplified the liver DNA damage in rats in case of NDMA endogenous synthesis.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Dose-Response Relationship, Drug; Neoplasms, Experimental; Nitrites; Nitrogen; Nitrosation; Sodium Nitrite

The luminal microenvironment including acid and nitric oxide (NO) has been implicated in Barrett's esophagus carcinogenesis. We investigated the ability of acid and NO to induce DNA damage in esophageal cells.. Transformed and primary Barrett's esophagus and adenocarcinoma cells were exposed to either acid, (pH 3.5), +/- antioxidant or NO from a donor or generated by acidification of nitrite in the presence of ascorbate +/- NO scavenger. Phosphorylation of histone H2AX and the neutral comet assay were used to detect DNA double-strand breaks (DSBs). Intracellular levels of reactive oxygen species and NO were detected with fluorescent dyes. Mitochondrial viability was measured with a rhodamine dye. Long-term survival was assessed by clonogenic assay.. Exposure to acid (pH 3.5) for > or =15 minutes induced DSBs in all cell lines (P < .05). There was a concomitant increase in intracellular reactive oxygen species in the absence of mitochondrial damage, and pretreatment with antioxidants inhibited DNA damage. Exposure to physiologic concentrations of NO produced from the NO donor or acidification of salivary nitrite induced DSBs in a dose- (>25 micromol/L) and cell-dependent manner (adenocarcinoma >Barrett's esophagus, P < .05). This occurred preferentially in S-phase cells consistent with stalled replication forks and was blocked with a NO scavenger. NO also induced DSBs in primary Barrett's esophagus cells treated ex vivo. Cells were able to survive when exposed to acid and NO.. Both acid and NO have the potential to generate DSBs in the esophagus and via distinct mechanisms.

Topics: Adenocarcinoma; Antioxidants; Ascorbic Acid; Barrett Esophagus; Cell Line, Tumor; Cell Survival; Cell Transformation, Neoplastic; Comet Assay; DNA Breaks, Single-Stranded; Dose-Response Relationship, Drug; Esophageal Neoplasms; Gastric Acid; Histones; Humans; Hydrazines; Hydrogen-Ion Concentration; Mitochondria; Nitric Oxide; Nitric Oxide Donors; Phosphorylation; Reactive Oxygen Species; S Phase; Sodium Nitrite; Time Factors

Combined effects of sodium nitrite (NaNO2) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) on liver, colon and Zymbal's gland carcinogenesis were assessed using a rat two-stage carcinogenesis model, with a focus on involvement of oxidative stress. Male 6-week-old F344 rats were given a single intraperitoneal injection of 200 mg/kg of diethylnitrosamine and 4 subcutaneous injections of 40 mg/kg of 1,2-dimethylhydrazine for initiation. Then, they were administered 0 or 300 ppm IQ in the diet or 0, 0.1 or 0.2% NaNO2 in their drinking water for 27 weeks. The treatment with NaNO2+IQ significantly enhanced colon and Zymbal's gland carcinogenesis and tended to enhance hepatocarcinogenesis. The incidence of lung tumors in the IQ-treated groups was significantly increased as compared with the initiation alone group. In a second experiment, male rats were given IQ or NaNO2 under the same conditions as before for 1 week, and at sacrifice, their liver and colon tissue or mucosa were collected for analysis of 8-hydroxydeoxyguanosine (8-OHdG), thiobarbituric acid reactive substances (TBARS), acrolein-modified protein and the bromodeoxyuridine-labeling index (BrdU-LI) (in the colon). In the colon, 8-OHdG, acrolein-modified protein levels and BrdU-LI were significantly increased by the combined treatment. These results indicate that the treatment with NaNO2 enhances IQ-induced colon and Zymbal's gland carcinogenesis in rats and that oxidative DNA damage and lipid peroxidation may partly be involved, especially in the colon. In addition, this experiment showed that IQ can act as a potent lung carcinogen in rats.

Topics: 1,2-Dimethylhydrazine; Administration, Oral; Alkylating Agents; Animals; Carcinogens; Cell Transformation, Neoplastic; Colonic Neoplasms; Diethylnitrosamine; DNA Damage; Drug Interactions; Indicators and Reagents; Injections, Subcutaneous; Lipid Peroxidation; Liver Neoplasms; Lung Neoplasms; Male; Oxidative Stress; Quinolines; Rats; Rats, Inbred F344; Sodium Nitrite

GGN-MRP is an extract from the Maillard reaction products of nitrite with glucose and glycine in the Maillard browning system. No genotoxicity of GGN-MRP in culture hepatocyte was found. A two-stage transformation protocol was used to transform chemically mouse embryo fibroblast C3H10T1/2 cells. To initiate transformation, the cells were treated with benzo[a]pyrene [B(a)P; 0.1 microg/mL], and GGN-MRP (0.01, 0.1, and 1.0 mg/mL) was employed to subsequently complete the transformation process. Malignant transformed foci were formed in B(a)P-initiated and GGN-MRP-promoted C3H10T1/2 cells after 8 weeks. Cells treated with GGN-MRP alone failed to induce transformation. However, cells initiated with B(a)P and promoted by GGN-MRP demonstrated oncogenic properties. Transformed colonies derived from GGN-MRP-treated cells exhibited enhanced growth rate, anchorage independence, and tumorgenicity in animals relative to parent cells. These results indicated that GGN-MRP contains a tumor promoter and may induce tumor promotion by two-stage oncogenesis.

Topics: Animals; Cell Division; Cell Line; Cell Transformation, Neoplastic; Glucose; Glycine; Maillard Reaction; Mice; Sodium Nitrite

N-Nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (NO-NTA) is a product of a model browning system in the presence of sodium nitrite. In this study, the chemical structure is confirmed by spectral studies, including UV, mass spectrometry, nuclear magnetic resonance and infrared spectroscopy. NO-NTA is strongly genotoxic to the rat hepatocyte and is moderately cytotoxic to mouse C3H10T1/2 cells. Results obtained in this study indicate that NO-NTA inflicted DNA damage through the formation of a DNA adduct. In addition, C3H10T1/2 cells were treated with NO-NTA and, following addition of 12-O-tetradecanoylphorbol-13-acetate (TPA) as promotor, the increase of transformed foci indicated that NO-NTA could possibly be an initiator [corrected] of TPA tumor promotion. A transformed cell line from NO-NTA initiated and TPA promoted foci increased saturation density and growth ability in soft agar reactive to the control line. These results suggest that the formation of a genotoxic agent of nitroso-derivatives may take place in a nitrite-containing food system during processing and cooking.

Topics: Animals; Cell Division; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Female; Glucose; Maillard Reaction; Mice; Mutagens; Nitrosamines; Rats; Sepharose; Sodium Nitrite; Tetrazolium Salts; Tyramine; Tyrosine

The addition of sodium nitrite (NaNO2; 5-20 mM) for 72 h to mouse BALB/c3T3 cells resulted in the induction of transformed foci (type III foci) in a dose-dependent manner. The cells isolated from the NaNO2-induced transformed foci produced progressively growing tumors when inoculated into nude mice subcutaneously at an inoculum size of 1 X 10(6) cells per site. In contrast, the original untreated cells did not take even at an inoculum size of 1 X 10(7) cells per site. The possibility that NaNO2 might react with cellular or medium components to make carcinogenic N-nitrosamines and that these might induce cell transformation was examined and almost excluded. Thus, nitrite itself seems to have a cell transforming activity. Recent evidence suggests that NO2- is produced by the activated macrophages of mammals. We also detected NO2- production in culture media in the mouse macrophage-like cell line J774-A1 after lipopolysaccharide (LPS) treatment, and also in the human promyeloleukemia cell line HL60 after differentiation into macrophage-like cells by 12-O-tetradecanoyl phorbol-13-acetate and further activation by LPS.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Culture Media; Dose-Response Relationship, Drug; Macrophages; Mice; Mice, Inbred BALB C; Nitrites; Nitrosamines; Sodium Nitrite

On the primary rat lung cell cultures, a study was made of the transforming action of sodium nitrite (NN) and amidopyrine combination with the control for formation of carcinogen--N-nitrosodimethylamine (NDMA) in growth medium. The manifestation of transformation was registered by appearance of morphological changes and multilayer growth foci. As criteria for evaluation after 48 hour treatments with NDMA, NN and AP, were used DNA-synthetizing activity of cells, mitotic index, frequency of mitoses pathology, monolayer density. The effects of transforming dose of NN alone and in combination with AP were the same. But malignization (tumor development in newborn rats in the points of cell suspension inoculation) took place only after administration of NN in combination with AP, when carcinogen was formed. Theophylline decreased the action of NN-AP combination.

Topics: Aminopyrine; Animals; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Cells, Cultured; Lung; Mitosis; Nitrites; Nitroso Compounds; Rats; Sodium Nitrite; Theophylline

The toxic and transformation actions of sodium nitrite (NN) were studied on the primary lung cell cultures of 3-day old noninbred rats. The monolayer density, mitotic index, DNA-synthetizing activity of cells, frequency of pathological mitoses changes in the character of growth, appearance of the multilayer growth foci serve as a criteria for evaluation. It is established that sodium nitrite has a transforming effect on lung cells and that the concentration of 1.035 mg/ml of nutrient medium is a transforming dose at 48 h exposition. Summation of the effect by prolongation of cell exposition with the NN prethreshold doses is possible.

Topics: Animals; Cell Survival; Cell Transformation, Neoplastic; Cells, Cultured; Lung; Mitosis; Nitrites; Rats; Sodium Nitrite

Topics: Aminopyrine; Animals; Azaguanine; Cell Transformation, Neoplastic; Cells, Cultured; Cricetinae; Drug Resistance; Embryo, Mammalian; Female; Mesocricetus; Mutation; Nitrites; Pregnancy; Sodium Nitrite

Hamster embryos were treated with various doses of NaNO2 in utero, by its oral administration to the mothers, and then the embryonic cells were examined for micronucleus formation, chromosomal aberrations, morphological or malignant transformation and drug-resistant mutations. For induction of resistant mutations, the cells were cultured in normal medium for 72 h, and then selected in media containing 8-azaguanine (10 or 20 microgram/ml) or 1 mM ouabain. This treatment with NaNO2 caused marked dose-dependent induction of 8-azaguanine- and ouabain-resistant mutations. Cultured embryonic fibroblasts in the resting state also showed a marked dose-dependent increase in micronucleus formation but not an increase in chromosomal aberrations. This treatment also caused morphological and neoplastic transformation of the cells. Transplacental oral treatment with DMN, as a positive control, caused changes of similar extent in biological effects of embryonic fibroblasts, and in addition it caused chromosomal aberrations in metaphase plates. On the contrary, transplacental oral application of NaNO2 did not induce any biological change in cultured embryonic fibroblasts.

Topics: Abnormalities, Drug-Induced; Animals; Azaguanine; Carcinogens; Cell Transformation, Neoplastic; Chromosome Aberrations; Cricetinae; Drug Evaluation, Preclinical; Drug Resistance; Embryo, Mammalian; Female; Maternal-Fetal Exchange; Mesocricetus; Mutagens; Nitrites; Ouabain; Pregnancy; Sodium Nitrite; Teratogens