Matsuoka A, Ozaki M, Takeshita K, Sakamoto H, Glatt H R, Hayashi M, Sofuni T
Division of Genetics and Mutagenesis, National Institute of Health Sciences, Tokyo, Japan.
Mutagenesis. 1997 Sep;12(5):365-72. doi: 10.1093/mutage/12.5.365.
We found that different ploidy effects were induced in four Chinese hamster-derived cell lines (V79-MZ, V79, CHL and CHO-K1) treated through two cell cycles with polycyclic aromatic hydrocarbons in the absence of a metabolic activation system. 5-Bromodeoxyuridine was used to investigate cell cycle delay and sister chromatid exchanges (SCE) induced by the chemicals. Benzo[a]pyrene (BP) induced aneuploidy at 2.5-10 micrograms/ml in V79-MZ cells. 7,12-Dimethylbenz[a]anthracene (DMBA) induced polyploidy at 3.125-6.25 and 6.25-1.25 micrograms/ml in V79-MZ and V79 cells respectively. Higher concentrations caused cell cycle delay and, therefore, did not affect ploidy. BP and DMBA did not induce a significant increase in SCE frequency at the above doses. 3-Methylcholanthrene tested up to its solubility limit (10 micrograms/ml) did not induce numerical aberrations in any cell line. The clastogen mitomycin C, tested up to 0.01 microgram/ml, did not produce numerical aberrations but did significantly increase SCE frequency in all cell lines. The spindle poison colchicine, tested up to 0.1 microgram/ml, induced ploidy changes in the four cell lines that showed different sensitivities. Four cell lines showed no arylhydrocarbon hydroxylase activity, and V79-MZ, but not the other cells lines, showed high glutathione S-transferase activity. Aneuploidy induction by BP and polyploidy induction by DMBA in the absence of S9 mix in vitro have not been described before, and the finding might be due to the effect on tubulin. Due to their specificity and high sensitivity, the V79-MZ and V79 cell lines might be good systems for detecting aneuploidogens.
我们发现，在没有代谢激活系统的情况下，用多环芳烃处理四个中国仓鼠来源的细胞系（V79-MZ、V79、CHL和CHO-K1）两个细胞周期，会诱导出不同的倍性效应。使用5-溴脱氧尿苷来研究化学物质诱导的细胞周期延迟和姐妹染色单体交换（SCE）。苯并[a]芘（BP）在2.5 - 10微克/毫升的浓度下可诱导V79-MZ细胞出现非整倍体。7,12-二甲基苯并[a]蒽（DMBA）在V79-MZ和V79细胞中分别在3.125 - 6.25微克/毫升和6.25 - 1.25微克/毫升的浓度下诱导多倍体形成。更高的浓度会导致细胞周期延迟，因此不会影响倍性。BP和DMBA在上述剂量下未诱导SCE频率显著增加。测试至其溶解度极限（10微克/毫升）的3-甲基胆蒽在任何细胞系中均未诱导染色体数目畸变。测试至0.01微克/毫升的致断裂剂丝裂霉素C未产生染色体数目畸变，但在所有细胞系中均显著增加了SCE频率。测试至0.1微克/毫升的纺锤体毒物秋水仙碱在四个表现出不同敏感性的细胞系中诱导了倍性变化。四个细胞系均未显示芳烃羟化酶活性，且V79-MZ细胞系（而非其他细胞系）显示出高谷胱甘肽S-转移酶活性。在体外无S9混合物的情况下，BP诱导非整倍体和DMBA诱导多倍体的现象此前未见报道，这一发现可能是由于对微管蛋白的影响。由于其特异性和高敏感性，V79-MZ和V79细胞系可能是检测非整倍体诱导剂的良好系统。
