COMPUTER ANALYSIS OF CARCINOGENESIS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

本项目的目的是研究 应用定量二维凝胶的致癌作用 电泳 该技术允许分离总的 细胞多肽在一个单一的凝胶，让我们检查 蛋白质模式的质和量的变化 合成A细胞经历肿瘤转化。 研究 重点是（1）计算机系统的持续发展 （称为ELSIE 4）用于自动分析凝胶和（2）使用 ELSIE 4用于分析需要计算机分析的实验， 二维凝胶 在过去的一年里，我们继续 开发软件工具，以帮助调查人员识别 有趣的地方。 用于搜索斑点的统计测试 在实验过程中可能会发生变化的方法已经被改进了， 新的测试，如启发式聚类，已经包括在内。 该系统目前在一些中型计算机上运行， 以及IBM PS/2 60型个人电脑。 ELSIE 4 该系统正在实验室的一些项目中使用， 包括一个旨在研究不同的 转化癌基因对大鼠肝脏蛋白质合成的影响 来源的上皮（RLE）细胞。 RLE细胞具有正常染色体 计数（2N = 42），染色体核型正常，除了 在1号染色体的q臂上复制 不同 含有转化癌基因的逆转录病毒，如v-ras，v- RAF和V-myc已被用于感染和转化这些细胞。 v-ras转化细胞已被单细胞克隆化。 所有 测试的转化细胞克隆在软琼脂中生长，并且高度 致瘤的 二维凝胶已经在正常和 转化的细胞和一些显著的变化， 多肽合成已经被注意到。

The objective of this project is to study the mechanism of carcinogenesis using quantitative two-dimensional gel electrophoresis. This technique allows the separation of total cellular polypeptides on a single gel and lets us examine both qualitative and quantitative changes in the pattern of protein synthesis a the cell undergoes neoplastic transformation. Research is focused on (1) continued development of the computer system (dubbed ELSIE 4) used to automatically analyze gels and (2) use of ELSIE 4 to analyze experiments requiring computerized analysis of two-dimensional gels. In the past year we have continued developing software tools to aid the investigator in identifying interesting spots. Statistical tests designed to search for spots that may vary over the course of an experiment have been refined, and new tests, such as heuristic clustering, have been included. The system is now running on a number of mid-sized computers as well as the IBM PS/2, model 60 personal computer. The ELSIE 4 system is being used in a number of projects in the Laboratory, including one designed to study the effects of different transforming oncogene on the synthesis of proteins in rat liver- derived epithelial (RLE) cells. RLE cells have a normal chromosome count (2N=42), the chromosomes appear karyotypically normal except for a duplication in q arm of chromosome #1. Different retroviruses containing transforming oncogenes, such as v-ras, v- raf, and v-myc have been used to infect and transform these cells. The v-ras-transformed cell have been single-cell cloned. All transformed cell clones tested grow in soft agar and are highly tumorigenic. Two-dimensional gels have been run on the normal and transformed cells and a number of significant variations in polypeptide synthesis have been noted.