Haynes J R, Downing J R
Department of Tumor Cell Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Mol Cell Biol. 1988 Jun;8(6):2419-27. doi: 10.1128/mcb.8.6.2419-2427.1988.
A contact-inhibited revertant of mink cells transformed by the Gardner-Arnstein strain of feline sarcoma virus was isolated by fluorescence-activated sorting of cells stained with the mitochondria-specific dye rhodamine 123. The revertant cell line exhibited a decrease in its proliferative rate and saturation density and a complete loss of its capacity for anchorage-independent growth, but it remained tumorigenic when inoculated into nude mice. The revertant cells retained a rescuable Gardner-Arnstein feline sarcoma provirus, expressed high levels of the v-fes oncogene product and its associated tyrosine kinase activity, manifested elevated levels of phosphotyrosine-containing cellular proteins similar to those observed in v-fes-transformed cells, and were refractory to retransformation by retroviruses containing the v-fes, v-fms, and v-ras oncogenes. Fusion of the revertant and parental cells generated somatic cell hybrids which formed colonies in semisolid medium, indicating that the block in transformation was recessive. These data together with the observation that the revertant phenotype is unstable in continuous culture suggest that the loss of transformation is due to the presence of limiting quantities of a gene product which functions downstream of the v-fes-coded kinase in the mitogenic pathway.
通过用线粒体特异性染料罗丹明123染色的细胞进行荧光激活分选,分离出了由加德纳 - 阿恩斯坦猫肉瘤病毒株转化的水貂细胞的接触抑制回复突变体。该回复突变体细胞系的增殖速率和饱和密度降低,并且完全丧失了其不依赖贴壁生长的能力,但是当接种到裸鼠中时它仍然具有致瘤性。回复突变体细胞保留了可拯救的加德纳 - 阿恩斯坦猫肉瘤前病毒,表达高水平的v - fes癌基因产物及其相关的酪氨酸激酶活性,表现出与在v - fes转化细胞中观察到的相似的含磷酸酪氨酸的细胞蛋白水平升高,并且对含有v - fes、v - fms和v - ras癌基因的逆转录病毒的再转化具有抗性。回复突变体和亲本细胞的融合产生了在半固体培养基中形成集落的体细胞杂种,表明转化中的阻断是隐性的。这些数据以及回复突变体表型在连续培养中不稳定的观察结果表明,转化的丧失是由于在有丝分裂途径中在v - fes编码的激酶下游起作用的基因产物数量有限所致。