Li G C, Ling C C, Endlich B, Mak J Y
Department of Radiation Oncology, University of California, San Francisco 94143.
Cancer Res. 1990 Aug 1;50(15):4515-21.
Rat embryo cells or Rat-1 fibroblasts were transfected with either an activated c-myc or a c-Ha-ras from the T24/EJ bladder carcinoma, or they were cotransfected with both. A gene conferring neomycin or hygromycin resistance was also cotransfected so that independent cell lines could be selected by growth in medium containing the antibiotic. Certain isolates from cells transfected with only one type of oncogene were further transformed by exposure to 600 cGy of 250-kVp X-rays. Successful transfection and transformation were characterized by altered morphology, increased plating efficiency, shorter doubling time, longer life span, foci formation, anchorage-independent growth, and Southern and Northern hybridization analysis. The thermal response of these cells at different stages of oncogenic transformation was examined by exposing exponentially growing cells to 45 degrees C for 0 to 45 min and measuring cellular survivals using colony formation assay. We found that cells transfected with myc oncogene, singly or in combination with ras, were more sensitive to thermal stress. Aside from that, the cells' thermal sensitivity was not affected by the degree or the nature of transformation.
将来自T24/EJ膀胱癌的活化c-myc或c-Ha-ras转染大鼠胚胎细胞或大鼠-1成纤维细胞,或者将二者共转染。同时共转染赋予新霉素或潮霉素抗性的基因,以便通过在含有抗生素的培养基中生长来选择独立的细胞系。仅用一种癌基因转染的细胞的某些分离株通过暴露于600 cGy的250-kVp X射线进一步转化。成功的转染和转化通过形态改变、接种效率提高、倍增时间缩短、寿命延长、集落形成、不依赖贴壁生长以及Southern和Northern杂交分析来表征。通过将指数生长的细胞暴露于45℃ 0至45分钟并使用集落形成试验测量细胞存活率,来检测这些细胞在致癌转化不同阶段的热反应。我们发现,单独或与ras共转染myc癌基因的细胞对热应激更敏感。除此之外,细胞的热敏感性不受转化程度或性质的影响。