TWO-DIMENSIONAL GEL ANALYSIS OF ONCOGENE-MEDIATED TRANSFORMATION

本项目的目的是研究癌症发生的机制 使用定量二维凝胶电泳（2DG）。 这 这项技术让我们可以检查质量和数量的变化， 当细胞经历一个过程时， 肿瘤性转化 研究的重点是（1）持续 开发用于分析凝胶的计算机系统（称为ELSIE），以及（2） 使用ELSIE分析需要计算机分析的实验 的2DG。 ELSIE系统正在实验室中用于研究 不同转化癌基因对蛋白质合成的影响。 大鼠肝上皮（RLE）细胞已被分离和单细胞培养。 克隆的 这些细胞具有正常的二倍体核型，并代表了一种 均质、克隆系统转化的研究。 不同 含有转化癌基因的逆转录病毒，如v-H-ras，v-N-ras， v-raf或v-myc/v-raf嵌合体已被用于转化RLE细胞或 人二倍体成纤维细胞系（LG-1）。 的多肽模式， 转化的细胞与其未转化的细胞显著不同， 祖先 虽然质量上的差异很少见，但超过30%的 斑点可以在数量上变化。 非常相似的癌基因，如v-H-ras 和v-N-ras，导致基本上不可区分的2DG模式。 这些 模式与由v-raf和v-myc/v-raf介导的模式不同 转型 v-raf和v-myc/v-raf模式也容易被识别。 彼此之间的区别，但更相似，而不是 未转化的或ras转化的细胞。

The objective of this project is to study the mechanism of carcinogenesis using quantitative two-dimensional gel electrophoresis (2DG). This technique lets us examine both qualitative and quantitative changes in the synthesis of thousands of cellular polypeptides as the cell undergoes neoplastic transformation. Research is focused on (1) the continued development of the computer system (dubbed ELSIE) to analyze gels and (2) the use of ELSIE to analyze experiments requiring computerized analysis of 2DGs. The ELSIE system is being used in the laboratory to study the effects of different transforming oncogenes on the synthesis of proteins. Rat liver epithelial (RLE) cells have been isolated and single-cell cloned. These cells have a normal diploid karyotype and represent a homogeneous, clonal system for the study of transformation. Different retrovirus containing transforming oncogenes, such as v-H-ras, v-N-ras, v-raf, or a v-myc/v-raf chimera have been used to transform RLE cells or a human diploid fibroblast line (LG-1). The polypeptide patterns of the transformed cells differ significantly from their non-transformed progenitors. While qualitative differences are rare, over 30% of the spots may vary quantitatively. Very similar oncogenes, such as v-H-ras and v-N-ras, cause essentially indistinguishable 2DG patterns. These patterns are distinct from those caused by v-raf and v-myc/v-raf mediated transformation. The v-raf and v-myc/v-raf patterns are also readily distinguishable, but are more similar to each other than to non-transformed or to ras transformed cells.