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首页> 外文期刊>Molecular and Cellular Biology >Establishment of two rabbit mammary epithelial cell lines with distinct oncogenic potential and differentiated phenotype after microinjection of transforming genes.
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Establishment of two rabbit mammary epithelial cell lines with distinct oncogenic potential and differentiated phenotype after microinjection of transforming genes.

机译：显微注射转化基因后，建立两种具有独特致癌潜能和分化表型的兔乳腺上皮细胞系。

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The goal of this work was to establish an assay for transformation of epithelial cells. Two epithelial cell lines were obtained after microinjecting transforming genes into primary rabbit mammary secretory cells. The cell lines were analyzed for their oncogenic potential and for the maintenance of a differentiated phenotype. A fully transformed cell line, which retained epithelial cell organization, was obtained by coinjecting simian virus 40 DNA and the activated human c-Ha-ras gene. The proliferation rate of these cells was high, with a doubling time of 16 h. Their growth was anchorage independent, and they had lost contact inhibition. The cells were tumorigenic in nude mice, but had no metastatic potential. Both microinjected DNAs were efficiently transcribed and translated, in contrast to the casein genes, which were expressed in primary cells but not in the transformed cell line. An immortalized cell line established after injection with simian virus 40 DNA alone was characterized by a moderate rate of proliferation with a doubling time of approximately 30 h. The growth of these cells was contact inhibited and anchorage dependent. The cells were not tumorigenic in nude mice. The viral DNA was expressed during early passages, as shown by the presence of the large T antigen in cell nuclei, but not at later passages. A high number of lactogenic hormone receptors were found associated with the cell surface. Despite the presence of these receptors, no induction of genes coding for milk proteins was observed after addition of prolactin. These data demonstrate that this assay system can be used to assess the immortalizing and transforming potential of candidate oncogenes in epithelial cells.
机译：这项工作的目的是建立一种转化上皮细胞的检测方法。将转化基因显微注射到原代兔乳腺分泌细胞中后，获得了两种上皮细胞系。分析细胞系的致癌潜能和分化表型的维持。通过同时注射猿猴病毒40 DNA和激活的人c-Ha-ras基因，获得了保留了上皮细胞组织的完全转化的细胞系。这些细胞的增殖速率很高，倍增时间为16小时。它们的生长与锚定无关，并且失去了接触抑制作用。这些细胞在裸鼠中具有致瘤性，但没有转移潜力。与酪蛋白基因相反，两种微量注射的DNA均能有效转录和翻译，酪蛋白基因在原代细胞中表达，而在转化细胞系中不表达。单独注射猿猴病毒40 DNA后建立的永生细胞系的特征在于适度的增殖，倍增时间约为30小时。这些细胞的生长受到接触抑制和锚定依赖性。这些细胞在裸鼠中没有致瘤性。病毒DNA在早期传代期间表达，如细胞核中大T抗原的存在所表明，但在后来的传代中没有表达。发现大量促乳激素受体与细胞表面有关。尽管存在这些受体，但添加催乳激素后未观察到编码乳蛋白的基因的诱导。这些数据表明，该测定系统可用于评估上皮细胞中候选致癌基因的永生化和转化潜能。

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