Ohmori Kiyomi, Sasaki Kiyoshi, Asada Shin, Tanaka Noriho, Umeda Makoto
Chemistry Division, Kanagawa Prefectural Institute of Public Health, 1-3-1 Shimomachiya, Chigasaki, Kanagawa 253-0087, Japan.
Mutat Res. 2004 Feb 14;557(2):191-202. doi: 10.1016/j.mrgentox.2003.10.014.
It has become an important task to develop a simple in vitro method for the detection of non-genotoxic carcinogens, among which tumor promoters are included. Bhas 42 cells are v-Ha-ras-transfected BALB/c 3T3 cells and are regarded as initiated cells in the 2-stage transformation paradigm. We designed a method for detecting tumor promoters by the use of Bhas 42 cells at advanced passage generation. In this method, the cells are cultured in six-well plates for 17 days during which test chemicals are added in the medium for 11 days from days 3 to 14. The end-point of the assay is the induction of transformed foci. When the tumor promoter TPA was used, a significant number of transformed foci were induced concentration-dependently, whereas only a few foci were observed in control cultures. When various chemicals were examined by the method, a reasonable correlation was observed with the reported tumor-promoting ability in animal experiments. We propose that the Bhas 42 cell transformation method is practical and useful for the detection of tumor promoters.
开发一种简单的体外检测非遗传毒性致癌物（包括肿瘤促进剂）的方法已成为一项重要任务。Bhas 42细胞是v-Ha-ras转染的BALB/c 3T3细胞，在两阶段转化模式中被视为起始细胞。我们设计了一种利用传代次数较多的Bhas 42细胞检测肿瘤促进剂的方法。在该方法中，将细胞接种于六孔板中培养17天，从第3天到第14天的11天时间里在培养基中加入受试化学物质。检测的终点是转化灶的诱导。当使用肿瘤促进剂佛波酯（TPA）时，可浓度依赖性地诱导出大量转化灶，而在对照培养物中仅观察到少数灶。当用该方法检测各种化学物质时，与动物实验中报道的肿瘤促进能力存在合理的相关性。我们认为Bhas 42细胞转化方法对于检测肿瘤促进剂是实用且有效的。
