myosmine and Adenocarcinoma

The incidence of esophageal adenocarcinoma is rapidly rising in Western populations. Gastroesophageal reflux disease (GERD) is thought to be one of the most important risk factors. However, the mechanisms by which GERD enhances tumor formation at the gastroesophageal junction are not well understood. Myosmine is a tobacco alkaloid which has also a wide spread occurrence in human diet. It is readily activated by nitrosation and peroxidation giving rise to the same hydroxypyridylbutanone-releasing DNA adducts as the esophageal carcinogen N'-nitrosonornicotine. Therefore, the genotoxicity of myosmine was tested in a human esophageal adenocarcinoma cell line (OE33). DNA damage was assessed by single-cell gel electrophoresis (Comet assay). DNA strand breaks, alkali labile sites and incomplete excision repair were expressed using the Olive tail moment (OTM). The Fapy glycosylase (Fpg) enzyme was incorporated into the assay to reveal additional oxidative DNA damage. DNA migration was determined after incubation of the cells for 1-24h. Under neutral conditions high myosmine concentrations of 25-50mM were necessary to elicit a weak genotoxic effect. At pH 6 genotoxicity was clearly enhanced giving a significant increase of OTM values at 5mM myosmine. Lower pH values could not be tested because of massive cytotoxicity even in the absence of myosmine. Co-incubation of 25 mM myosmine with 1mM H(2)O(2) for 1h significantly enhanced the genotoxicity of H(2)O(2) but not the oxidative lesions additionally detected with the Fpg enzyme. In the presence of the peroxynitrite donor 3-morpholinosydnonimine (SIN-1) a dose-dependent significant genotoxic effect was obtained with 1-10mM myosmine after 4h incubation. NS-398, a selective inhibitor of cyclooxygenase 2, did not affect the SIN-1 stimulated genotoxicity of myosmine. Finally, the 23 h repair of N-methyl-N'-nitro-N-nitrosoguanidine-induced DNA lesions was significantly inhibited in the presence of 10mM myosmine. In conclusion, myosmine exerts significant genotoxic effects in esophageal cells under conditions which may prevail in GERD such as increased oxidative and nitrosative stress resulting from chronic inflammation.

Topics: Adenocarcinoma; Alkaloids; Cell Line, Tumor; Comet Assay; DNA Damage; DNA-Formamidopyrimidine Glycosylase; Esophageal Neoplasms; Humans

Myosmine has been regarded as a specific tobacco alkaloid until investigations pointed out that nuts and nut products constitute a significant source of myosmine. In the present study it is shown that the occurrence of myosmine is widespread throughout a large number of plant families. Using a method for extraction practicable for all examined foods, quantitative analysis through internal standard addition showed nanograms per gram amounts. Positively tested edibles were staple foods such as maize, rice, wheat flour, millet, potato, and milk and also cocoa, popcorn, tomato, carrot, pineapple, kiwi, and apples. No myosmine was detectable in other vegetables and fruits such as lettuce, spinach, cucumber, onion, banana, tangerines, and grapes. Myosmine is easily nitrosated giving rise to a DNA adduct identical to the esophageal tobacco carcinogen N-nitrosonornicotine. Therefore, the role of dietary myosmine in esophageal adenocarcinoma should be further investigated.

Topics: Adenocarcinoma; Alkaloids; Animals; Edible Grain; Esophageal Neoplasms; Fruit; Gas Chromatography-Mass Spectrometry; Milk; Vegetables

The tobacco alkaloid myosmine was detected in nut and nut products [J. Agric. Food Chem. 46 (1998) 2703]. Upon nitrosation, myosmine yields 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) and N-nitrosonornicotine (NNN) [J. Agric. Food Chem. 48 (2001) 392]. NNN is a strong oesophageal carcinogen in rats. Metabolic activation of NNN leads to formation of DNA and protein adducts which release HPB upon hydrolysis. In the present study the time, pH and dose-dependent nitrosation of myosmine and its covalent binding to DNA was investigated. [5-(3)H]myosmine was incubated with nitrite for 1-24 h in buffer solutions adjusted to pH 1-6. At pH 2-4 myosmine was easily nitrosated and gave rise to two major products, HPB and NNN, and five minor not yet identified products. Maximal formation was achieved for HPB at pH 2 after 8 h (72% of total radioactivity) and for NNN at pH 3 after 8 h (16%). For DNA binding studies labeled myosmine was incubated under nitrosation conditions with calf thymus DNA. Within 3 h up to 0.1% of the radioactivity was covalently bound to DNA. Endogenous nitrosation of myosmine, present in nuts and other dietary components could constitute a significant risk factor for tumours in the upper intestinal tract such as oesophageal adenocarcinoma, which are unrelated to tobacco smoking and alcohol abuse.

Topics: Adenocarcinoma; Alkaloids; Butanones; Chromatography, High Pressure Liquid; DNA Adducts; Dose-Response Relationship, Drug; Esophageal Neoplasms; Humans; Hydrogen-Ion Concentration; Nicotiana; Nitrosamines; Nitrosation; Nuts; Pyridines; Risk Factors; Time Factors