COMPUTER ANALYSIS OF CARCINOGENESIS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

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在过去的一年中， 合成数以千计的细胞。多肽作为细胞经历 肿瘤性转化研究的重点是（1）持续发展 用于自动分析凝胶的计算机系统（称为ELSIE 4） 和（2）使用ELSIE 4分析需要计算机处理的实验 二维凝胶分析。在过去的一年里，我们的实验室 经历了一个重大的变化，现在包括四个太阳微系统计算机 工作站，在局域网中联网在一起。重大进展 是为了让系统不再使用专门的硬件 该系统已成功地移植到许多不同的计算机上。 统计测试，旨在寻找可能在不同的地点， 一个实验的过程已经被改进了。一旦有趣的景点 标记后，可以使用标准图像处理软件对其进行检查 在这个实验室里开发的。ELSIE 4系统用于 实验室研究不同转化癌基因对 蛋白质的合成。大鼠肝上皮（RLE）细胞已被分离 单细胞克隆这些细胞具有正常的二倍体核型， 代表了一个同质的，克隆系统的研究转化。 不同的逆转录病毒含有转化癌基因，如v-H-ras， V-raf和V-myc已被用于转化这些细胞。所有 v-H-ras转化的克隆在软琼脂中生长并且是高度致瘤性的。一 已经注意到多肽合成中的许多显著变化 在转化和非转化细胞的2DG模式之间。进一步 研究正在进行中，以确定蛋白质是否存在共同的模式， 其合成被不同的致癌基因改变。一系列类似的 已经进行了利用人成纤维细胞的实验。

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) 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, our laboratory has undergone a major change and consists now of four Sun Microsystems computer workstations, networked together in a local area network. Major progress has been made in weaning the system from the use of specialized hardware. The system has been successfully ported to a number of different computers. Statistical tests designed to search for spots that may vary over the course of an experiment have been refined. Once interesting spots are flagged, they may be examined using standard image processing software developed in this laboratory. The ELSIE 4 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 normal diploid karyotype and represent a homogeneous, clonal system for the study of transformation. Different retroviruses containing transforming oncogenes, such as v-H-ras, v-raf, and v-myc, have been used to transform these cells. All v-H-ras-transformed clones grow in soft agar and are highly tumorigenic. A number of significant variations in polypeptide synthesis have been noted between the 2DG patterns of transformed and non-transformed cells. Further studies are underway to determine if a common pattern exists for proteins whose synthesis are altered by the different oncogenes. A similar series of experiments, utilizing human fibroblasts, has been undertaken.