Yasunaga Katsuaki, Kiyonari Akiko, Oikawa Takeshi, Abe Naoki, Yoshikawa Kunie
Genetic Toxicology Group, Toxicology Division II, Mitsubishi Chemical Safety Institute Ltd., Ibaraki 314-0255, Japan.
Environ Mol Mutagen. 2004;44(4):329-45. doi: 10.1002/em.20053.
There is a need for simple rapid tests for evaluating the carcinogenic potential of the thousands of chemical compounds that are developed each year. The DNA-damaging effects of 83 National Toxicology Program (NTP) chemicals, including noncarcinogens and carcinogens, were examined in the umu test by measuring the expression of the umuDC-lacZ genes in Salmonella typhimurium TA1535/pSK1002. Salmonella were exposed to individual NTP chemicals at 37 degrees C for 2 hr both with and without a rat liver S9 mix; the treated cells were then diluted and incubated for a further 2 hr (posttreatment assay). O-nitrophenyl-beta-D-galactoside was added to the cultures and the beta-galactosidase activity driven by the Salmonella umuDC-lacZ genes was determined by measurement of the OD(420 nm) and OD(550 nm) of the cultures. Salmonella cell number was simultaneously determined by measurement of OD(600 nm). The overall concordance between genotoxicity in the umu test and carcinogenicity was 67%, which was similar to the concordance between Ames' test results and carcinogenicity (63%) using the same 83 NTP chemicals. The results of this study indicate that the umu test with a single Salmonella strain is a simple rapid system, with accuracy comparable to existing, more time-consuming assays.
每年都会研发出数千种化合物,因此需要简单快速的检测方法来评估它们的致癌潜力。通过测量鼠伤寒沙门氏菌TA1535/pSK1002中umuDC-lacZ基因的表达,在umu试验中检测了83种美国国家毒理学计划(NTP)化学品的DNA损伤效应,这些化学品包括非致癌物和致癌物。将沙门氏菌在37摄氏度下分别暴露于单一的NTP化学品中2小时,一组添加大鼠肝脏S9混合物,另一组不添加;然后将处理过的细胞稀释并进一步培养2小时(处理后测定)。向培养物中加入邻硝基苯基-β-D-半乳糖苷,并通过测量培养物的OD(420nm)和OD(550nm)来测定由沙门氏菌umuDC-lacZ基因驱动的β-半乳糖苷酶活性。同时通过测量OD(600nm)来确定沙门氏菌细胞数量。umu试验中的遗传毒性与致癌性之间的总体一致性为67%,这与使用相同的83种NTP化学品时艾姆斯试验结果与致癌性之间的一致性(63%)相似。这项研究的结果表明,使用单一沙门氏菌菌株的umu试验是一个简单快速的系统,其准确性与现有的耗时更长的检测方法相当。