Miller C L, Fuseler J W, Brinkley B R
Cell. 1977 Sep;12(1):319-31. doi: 10.1016/0092-8674(77)90210-0.
Patterns of cytoplasmic microtubules in somatic cell hybrids between transformed mouse cells and nontransformed human skin fibroblasts were examined using antitubulin antibodies as an immunofluorescent probe. Nontransformed cells have been shown to exhibit an extensive cytoplasmic microtubule complex (CMTC), while in transformed cells, this complex is greatly diminished. The hybrid populations contained both types of cells. In addition, they contained cells with previously undescribed intermediate CMTC phenotypes. The percentage of each phenotype present in hybrid populations was determined for sixteen hybrid clones. Seven clones were found which appeared transformed on the basis of their CMTC pattern. The others were comprised of various proportions of all the cell types described. Repeated quantitation of the proportions of these types in the hybrid populations showed them to be stable with time in culture. Growth in vitro of the hybrid clones was assayed by determining their saturation densities, their plating efficiencies on plastic and their colony-forming abilities in soft agar. In vitro growth of a cell population was found to be directly proportional to the percentage of cells in the population which showed the greatly diminished CMTC pattern which has been described for transformed cells. This is strong evidence that a greatly reduced CMTC is associated with transformed behavior, especially the increased capacity of transformed cells for in vitro growth.
利用抗微管蛋白抗体作为免疫荧光探针,检测了转化的小鼠细胞与未转化的人皮肤成纤维细胞之间的体细胞杂种中的细胞质微管模式。已证明未转化的细胞表现出广泛的细胞质微管复合体(CMTC),而在转化细胞中,这种复合体大大减少。杂种群体中包含这两种类型的细胞。此外,它们还包含具有先前未描述的中间CMTC表型的细胞。确定了16个杂种克隆中杂种群体中每种表型的百分比。发现7个克隆根据其CMTC模式似乎已转化。其他克隆由所述所有细胞类型的不同比例组成。对杂种群体中这些类型的比例进行反复定量,结果表明它们在培养过程中随时间保持稳定。通过测定杂种克隆的饱和密度、它们在塑料上的接种效率以及它们在软琼脂中的集落形成能力,检测了杂种克隆的体外生长。发现细胞群体的体外生长与群体中显示出已描述的转化细胞中大大减少的CMTC模式的细胞百分比成正比。这有力地证明,大大减少的CMTC与转化行为有关,特别是与转化细胞体外生长能力的增加有关。