Tissue and cell specific methylation, repair and synthesis of DNA in the upper gastrointestinal tract of Wistar rats treated with N-methyl-N'-nitro-N-nitrosoguanidine via the drinking water.

Abstract

Several potential cancer risk factors have been monitored concurrently in the upper gastrointestinal tract of young male Wistar rats given N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) via the drinking water, a regimen that induces a high yield of tumours in the pylorus and to a lesser extent in the duodenum. Radioimmunoassay was used to determine the amounts of O6-methyl-2'-deoxyguanosine (O6-MedG) formed in the tissue DNA of rats given MNNG at doses of 40 or 80 micrograms/ml for periods of 3, 6 and 12 weeks. The highest adduct concentration was found in the pylorus with progressively lower concentrations in the corpus and duodenum, jejunum, forestomach and oesophagus. Between 3 and 12 weeks these adduct levels decreased in all tissues and there was no evidence of a dose dependent accumulation of O6-MedG. When analysed by immunohistochemistry the distribution of cells with nuclei containing O6-MedG was seen to be heterogeneous in the various tissues. O6-Alkylguanine-DNA alkyltransferase activity increased during the 12 weeks of MNNG treatment in oesophagus and forestomach, but decreased to approximately 50% of the initial value in the corpus, pylorus, duodenum and jejunum. The major changes in DNA synthesis and cell proliferation were the marked upward expansion (i.e. towards the lumen) of the zone of replicating cells in the glands of the pylorus and the greatly increased numbers of replicating damaged cells (i.e. cells that contained O6-MedG whilst undergoing DNA synthesis) as determined by sequential immunohistochemical analysis and autoradiography. Such cells are the probable target cells in this chronic dose carcinogenesis regime. Although similar changes also occurred in the glands of the corpus these were of lesser extent and the changes of labelling index in the oesophagus and forestomach were relatively minor. In the duodenum, MNNG treatment led to erosion of the upper part of the glands so that the zone of cells containing O6-MedG overlapped with the zone of proliferating cells resulting in the formation of many replicating damaged cells. Thus, as in the single dose study (see preceding paper) the distribution of replicating damaged cells coincides with the tumour yield in the tissues of the upper gastrointestinal tract. As in the case of single doses of MNNG the risk factors for carcinogenesis are, a significant level of DNA damage, a lower capacity for DNA repair and an increased DNA synthetic activity, again suggesting that carcinogenic risk cannot readily be determined by studying risk factors individually.