COMPUTER ANALYSIS OF CARCINOGENESIS BY TWO-DIMENSIONAL GEL ELECTROPHORESIS

本课题的主要目的是研究 使用定量二维凝胶电泳的致癌作用。 该技术允许同时分离总细胞 多肽的单一聚丙烯酰胺凝胶，并允许我们检查 蛋白质模式的质的变化以及量的变化 在单个多肽中，因为细胞经历恶性转化。 目前的研究主要集中在以下几个方面：（1）继续 开发和改进计算机系统（称为ELSIE III） 目前用于自动分析二维凝胶;和（2） 应用ELSIE III分析了克隆变异， 转化细胞和细胞蛋白质模式的时间变化 在类固醇驱动的小鼠成纤维细胞转化期间， MMTV：v-ras嵌合体。 在过去的一年里，我们改进了现场发现 程序和开发的技术，以找到不同的斑点， 在实验过程中，定量或定性。 的 ELSIE III的数据组织已经修改， 可以比较不同实验产生的凝胶。 我们有 研究了9个单细胞克隆的多肽模式，这些克隆来自于 克隆细胞系H-4-II-E。 在982种蛋白质中， 亚克隆，只有5个蛋白质模式的定性变化， 检测到 然而，大约20%的蛋白质是在 数量上不同。 最后，我们检查了多肽 NIH 3 T3细胞克隆的模式，433. 这些细胞含有v-ras 并且可以被地塞米松诱导合成ras-p21 蛋白质并表达转化的表型。 正在努力 鉴定这些凝胶上的p21蛋白产物，并使用ELSIE III， 寻找定性和定量的蛋白质模式， 与转化表型相关。

The main objective of this project is to study the mechansim of carcinogenesis using quantitative two-dimensional gel electrophoresis. This technique allows for the simultaneous separation of total cellular polypeptides on a single polyacrylamide gel and allows us to examine qualitative changes in the protein patterns as well as quantitative changes in individual polypeptides as the cell undergoes malignant transformation. Research is, at present, focused on the following areas: (1) continued development and improvements on the computer system (dubbed ELSIE III) currently used to automatically analyze two-dimensional gels; and (2) application of ELSIE III to analyze both the clonal variation among transformed cells and the temporal changes in cellular protein patterns during steroid driven transformation of mouse fibroblasts transfected with MMTV:v-ras chimeras. In the past year we have improved the spot finding program and developed techniques to find spots which vary, either quantitatively or qualitatively, during the course of an experiment. The organization of data on ELSIE III has been modified so that it is now possible to compare gels generated by different experiments. We have examined the polypeptide patterns of 9 single cell clones derived from a clonal cell line, H-4-II-E. Out of 982 proteins found amond the various subclones, only 5 qualitative changes in the protein patterns were detected. About 20% of the proteins, however, were synthesized at quantitatively different rates. Finally, we have examined the polypeptide pattern of a clone of NIH3T3 cells, 433. These cells contain the v-ras oncogene and can be induced, by dexamethasone, to synthesize the ras-p21 protein and express the transformed phenotype. Efforts are underway to identify the p21 protein product on these gels and, using ELSIE III, to search for both qualitative and quantitative protein patterns that are associated with the transformation phenotype.