Helbig R, Gerland E, Zdzienicka M Z, Speit G
Universität Ulm, Abteilung Klinische Genetik, Germany.
Mutat Res. 1995 May;336(3):307-16. doi: 10.1016/0921-8777(95)00005-5.
The Chinese hamster cell line V-E5 is a mutant cell line isolated from V79 cells. The phenotypic characteristics of V-E5 strongly resemble those of cells from patients suffering from the genomic instability syndrome ataxia telangiectasia. In order to further characterize the mutant cell line and to get insight into the underlying genetic defect we compared the clastogenic and mutagenic effects of neocarzinostatin (NCS) and methyl methanesulfonate (MMS) in V-E5 and V79 wild-type cells (V79-LE). V-E5 cells were 2-3 times more sensitive to the cytotoxic effect of NCS or MMS. The clastogenic action of NCS was characterized by the predominant induction of chromosome breaks and dicentrics in both cell lines, whereas MMS mainly induced chromatid-type aberrations. The frequency of mutations induced by NCS as well as MMS was slightly enhanced in V-E5 cells compared to V79 cells treated with the same dose. However, the mutant cell line was found to be hypomutable when considering the same survival level as in the parental cell line. Molecular analysis of mutants induced by NCS revealed a high frequency of total deletions of the hprt gene in both cell lines. In contrast, among MMS-induced mutations only 11% deletion mutations were found in V79-LE, whereas in V-E5 MMS-induced deletions were seen in 52% of the hprt-deficient mutants. These results are discussed with respect to a possible relation between genomic instability, cell cycle control and mutational spectra.
中国仓鼠细胞系V-E5是从V79细胞中分离出来的突变细胞系。V-E5的表型特征与患有基因组不稳定综合征共济失调毛细血管扩张症患者的细胞非常相似。为了进一步表征该突变细胞系并深入了解潜在的遗传缺陷,我们比较了新制癌菌素(NCS)和甲磺酸甲酯(MMS)对V-E5和V79野生型细胞(V79-LE)的致断裂和诱变作用。V-E5细胞对NCS或MMS的细胞毒性作用的敏感性高2至3倍。NCS的致断裂作用在两种细胞系中均以染色体断裂和双着丝粒的主要诱导为特征,而MMS主要诱导染色单体型畸变。与用相同剂量处理的V79细胞相比,NCS和MMS诱导的V-E5细胞中的突变频率略有增加。然而,当考虑与亲代细胞系相同的存活水平时,发现该突变细胞系的诱变率较低。对NCS诱导的突变体进行分子分析发现,两种细胞系中hprt基因完全缺失的频率都很高。相比之下,在MMS诱导的突变中,V79-LE中仅发现11%的缺失突变,而在V-E5中,52%的hprt缺陷突变体中出现了MMS诱导的缺失。针对基因组不稳定、细胞周期控制和突变谱之间的可能关系对这些结果进行了讨论。