Ohsawa K, Furihata C, Mori M, Ikui E
Department of Molecular Oncology, University of Tokyo, Japan.
Mutat Res. 1993 Jun;287(2):307-19. doi: 10.1016/0027-5107(93)90023-9.
Male F344 rats were given test chemicals orally, and samples of their pyloric mucosa were incubated in vitro. Induction of unscheduled DNA synthesis (UDS) and stimulation of replicative DNA synthesis in the pyloric mucosa were then examined by addition of [3H]thymidine and simultaneous determinations of DNA synthesis in the presence and absence of hydroxyurea, an inhibitor of replicative DNA synthesis. DNA damage was also examined by the alkaline elution method with DNA single-strand scission as a marker. The results showed four types of abilities of the chemicals to affect UDS and replicative DNA synthesis in the pyloric mucosa of rat stomach 1-2 h after their administration: (1) induction of UDS and stimulation of replicative DNA synthesis by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), a glandular stomach carcinogen, (2) induction of only UDS by 4-nitroquinoline 1-oxide (4NQO), a glandular stomach carcinogen, (3) stimulation of only replicative DNA synthesis by NaCl, a glandular stomach tumor promoter, and (4) neither induction of UDS nor stimulation of replicative DNA synthesis by dimethylnitrosamine (DMN), a liver carcinogen. DNA single-strand scission was induced by MNNG and 4NQO, being maximal 2 h after their administration, but was not induced by NaCl or DMN. Thus it correlated well with the induction of UDS. The present results indicate four types of inductive abilities of chemicals on UDS and replicative DNA synthesis in rat stomach pyloric mucosa and show that this method can detect differences in the action mechanisms and organ specificities of glandular stomach carcinogens.
给雄性F344大鼠经口给予受试化学物质,然后将其幽门黏膜样本进行体外培养。接着通过添加[3H]胸腺嘧啶核苷并同时测定在有和没有羟基脲(一种复制性DNA合成抑制剂)存在的情况下的DNA合成,来检测幽门黏膜中不定时DNA合成(UDS)的诱导情况以及复制性DNA合成的刺激情况。还通过以DNA单链断裂为标志物的碱性洗脱法检测DNA损伤。结果显示,在给予化学物质1 - 2小时后,这些化学物质对大鼠胃幽门黏膜中UDS和复制性DNA合成有四种影响能力:(1)腺胃致癌物N - 甲基 - N'- 硝基 - N - 亚硝基胍(MNNG)诱导UDS并刺激复制性DNA合成,(2)腺胃致癌物4 - 硝基喹啉1 - 氧化物(4NQO)仅诱导UDS,(3)腺胃肿瘤促进剂氯化钠仅刺激复制性DNA合成,(4)肝癌致癌物二甲基亚硝胺(DMN)既不诱导UDS也不刺激复制性DNA合成。MNNG和4NQO诱导了DNA单链断裂,在给药后2小时达到最大值,但氯化钠或DMN未诱导。因此,它与UDS的诱导情况密切相关。目前的结果表明化学物质对大鼠胃幽门黏膜中UDS和复制性DNA合成有四种诱导能力,并表明该方法可以检测腺胃致癌物作用机制和器官特异性的差异。