COMPUTER ANALYSIS OF CARCINOGENESIS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

本课题的目的是为了研究黄褐斑的发病机制。 定量二维凝胶的致癌作用 电泳法。这项技术允许分离总的 单个凝胶上的细胞多肽，让我们研究这两个 蛋白质结构的质变和量变 合成a细胞经历肿瘤转化。研究 侧重于(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员识别 有趣的地方。为搜索斑点而设计的统计测试 在实验过程中可能会有所不同， 新的测试，如启发式聚类，已被包括在内。 该系统现在在一些中型计算机上运行，如 以及IBM PS/2，型号60的个人计算机。《埃尔西4》 该系统正在实验室的一些项目中使用， 其中包括一项旨在研究不同 转化癌基因对大鼠肝脏蛋白质合成的影响 来源上皮(RLE)细胞。RLE细胞有正常的染色体 计数(2N=42)，核型正常，除 1号染色体Q臂上的重复。不同 含有转化癌基因的逆转录病毒，如v-ras，v-ras RAF和v-myc被用来感染和转化这些细胞。 经v-ras基因转化的细胞已被单细胞克隆。全 测试的转化细胞克隆在软琼脂中生长，并高度 致癌的。二维凝胶已经在正常和 转化的细胞和一些重要的变化 多肽的合成已被注意到。

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.