Landolph J R, Fournier R E
Mutat Res. 1983 Feb;107(2):447-63. doi: 10.1016/0027-5107(83)90183-5.
We previously developed a quantitative assay for measuring the induction of ouabain-resistant (Ouar) variants in transformable C3H/20T1/2 Cl 8 mouse fibroblasts following treatment of the cells with chemical carcinogens. To further define the nature of the Ouar phenotype, we conducted microcell-mediated chromosome transfer studies using Ouar cell lines induced by chemical carcinogens in C3H/10T1/2 Cl 8 cells as donors and 8-azaguanine-resistant (Azgr) derivatives of the human cell lines, D98/AH2 and HT 1080, as recipients. Microcells prepared from one spontaneous and two carcinogen-induced Ouar mouse cell lines were able to transfer resistance to 0.01 and 1 mM Oua to ouabain-sensitive D98 and HT 1080 cells. The frequency of microcell hybrid formation ranged from 10(-6) to 10(-5). Karyotypic analysis of the microcell hybrids indicated that the Ouar phenotype of C3H/10T1/2 Cl 8 derivatives mapped to mouse chromosome 3, the chromosome to which the wild-type murine Oua-1 allele had previously been assigned. These studies show that both spontaneous and chemically induced high level Ouar phenotypes of C3H/10T1/2 Cl 8 mouse fibroblasts can be transferred via microcell-mediated chromosome transfer, and provide strong genetic evidence that chemically induced Ouar phenotypes of C3H/10T1/2 Cl 8 cells arise from mutations at Oua-1. In addition, this study sufficiently standardizes microcell-mediated chromosome transfer in the C3H/10T1/2 Cl 8 cell line so that this technique can be used to investigate the nature of other phenotypic changes in these cells, such as the chemically transformed phenotype.
我们之前开发了一种定量检测方法，用于测量在用化学致癌物处理可转化的C3H/20T1/2 Cl 8小鼠成纤维细胞后，哇巴因抗性（Ouar）变体的诱导情况。为了进一步明确Ouar表型的性质，我们进行了微细胞介导的染色体转移研究，使用在C3H/10T1/2 Cl 8细胞中由化学致癌物诱导产生的Ouar细胞系作为供体，以及人细胞系D98/AH2和HT 1080的8-氮杂鸟嘌呤抗性（Azgr）衍生物作为受体。从一个自发产生的和两个由致癌物诱导产生的Ouar小鼠细胞系制备的微细胞，能够将对0.01和1 mM哇巴因的抗性转移到对哇巴因敏感的D98和HT 1080细胞中。微细胞杂种形成的频率范围为10^(-6)至10^(-5)。对微细胞杂种的核型分析表明，C3H/10T1/2 Cl 8衍生物的Ouar表型定位于小鼠3号染色体，野生型小鼠Oua-1等位基因先前已被定位到该染色体上。这些研究表明，C3H/10T1/2 Cl 8小鼠成纤维细胞的自发和化学诱导的高水平Ouar表型都可以通过微细胞介导的染色体转移进行转移，并提供了有力的遗传学证据，证明C3H/10T1/2 Cl 8细胞的化学诱导Ouar表型源于Oua-1处的突变。此外，本研究充分规范了C3H/10T1/2 Cl 8细胞系中微细胞介导的染色体转移，以便该技术可用于研究这些细胞中其他表型变化的性质，如化学转化表型。
