Zhang H, Borman H D, Myhr B C
Genetic and Molecular Toxicology, Covance Laboratories Inc., 9200 Leesburg Pike, Vienna, VA 22182, USA.
Mutat Res. 2004 Apr 14;548(1-2):1-7. doi: 10.1016/j.mrfmmm.2003.05.002.
Syrian hamster embryo (SHE) cell transformation has been used for many years to study chemical carcinogenesis in vitro. It has been shown that this assay is probably the most predictive short-term test system for identifying rodent carcinogens. Although most of the operational difficulties encountered in the early stage of application of this assay have been overcome by culturing the SHE cells under slightly acidic conditions (pH 6.7), a relatively low level of induction of morphological transformation (MT) by known carcinogens still occurs for many cell isolates. In order to improve the response of this assay system to known carcinogens, the effect of incubation time of target SHE cells on the frequency of morphological transformation induced by benzo(a)pyrene (BaP) was investigated. It was shown that the morphological transformation frequency induced by BaP increased significantly (1.4-2.5-fold) when the incubation time of target cells was reduced from the usual 24h to less than 6h prior to seeding onto feeder layers. This improvement in sensitivity was consistent for different cell isolates. In addition, the enhanced response appeared to be a property of carcinogens because treatment with two non-carcinogens, l-ascorbic acid and 4-nitro-o-phenylenediamine, did not induce significant increases in the transformation frequency under the shortened incubation period for target cells. These results suggest that the response of the SHE cell transformation assay may be improved by optimizing the incubation time of the target SHE cells. In addition, the results of the present study provide further evidence to support the idea that morphological transformation of SHE cells results from a block of cellular differentiation of stem or stem-like cells.
叙利亚仓鼠胚胎(SHE)细胞转化试验多年来一直用于体外研究化学致癌作用。已表明该试验可能是鉴定啮齿动物致癌物最具预测性的短期检测系统。尽管通过在略酸性条件(pH 6.7)下培养SHE细胞克服了该试验早期应用中遇到的大多数操作困难,但对于许多细胞分离株而言,已知致癌物诱导的形态转化(MT)水平仍然相对较低。为了提高该检测系统对已知致癌物的反应,研究了靶SHE细胞孵育时间对苯并(a)芘(BaP)诱导的形态转化频率的影响。结果表明,在接种到饲养层之前,将靶细胞的孵育时间从通常的24小时减少到少于6小时,BaP诱导的形态转化频率显著增加(1.4 - 2.5倍)。这种敏感性的提高对不同的细胞分离株是一致的。此外,增强的反应似乎是致癌物的特性,因为用两种非致癌物L-抗坏血酸和4-硝基邻苯二胺处理,在靶细胞孵育期缩短的情况下,并未诱导转化频率显著增加。这些结果表明,通过优化靶SHE细胞的孵育时间,可以提高SHE细胞转化试验的反应。此外,本研究结果提供了进一步的证据支持SHE细胞的形态转化是由干细胞或干细胞样细胞的细胞分化受阻引起的这一观点。