GENE REGULATION IN BREAST, COLON AND SKIN CANCER

正常细胞转化为恶性肿瘤是一个多步骤的过程 这一过程是由基因改变和环境损害造成的。 大多数恶性细胞具有通过肿瘤转移的能力。 细胞脱离原发肿瘤，内渗，外渗 以及在远端器官中建立次级病灶。细胞外 基质（ECM）和基底膜由多种 组织和器官周围的蛋白质， 积极的细胞流动性的障碍。 转移过程包括 游走的恶性细胞和大分子之间的相互作用 基底膜和细胞外基质， 并通过增加蛋白水解酶的合成获得出口 内切酶 各种调查显示， ECM蛋白在各种恶性细胞中的合成。 的假设 这一假设是ECM的转录调节发生了改变， 蛋白质合成作为对转化状态的直接响应，并且 这种改变增强了转移。在本提案中，我们提出了一个 系统的方法来探索这个假设。 一组成对的 细胞系和肿瘤标本（正常细胞、息肉、原发性肿瘤和 转移性样品）进行分析 北方、南方和狭缝印迹技术， 编码骨连接蛋白、核心蛋白聚糖、I型胶原蛋白、 纤连蛋白和各种癌基因。管家蛋白β-肌动蛋白 和甘油磷酸脱氢酶，将作为这些 实验 核提取、DNA足迹、凝胶迁移率 并进行定点诱变试验以确定 改变转录的分子机制。 数据从这些 研究可以提供一种机制，通过这种机制来确定 肿瘤生长特定阶段的转移，并提供分子水平 标记以确定是否已经发生微转移。

Transformation of normal cells to malignant tumors is a multi-step process which results from genetic alterations and environmental insults. Most malignant cells possess the ability to metastasize via the detachment of cells form the primary tumor, intravasation, extravasation and establishment of secondary foci in distal organs. Extracellular matrices (ECM) and basement membranes are composed of a variety of proteins that surround tissues and organs and present impermeable barriers to active cell mobility. The metastatic process involves interactions between wandering malignant cells and macromolecules of basement membranes and extracellular matrices, whereby the cells attach to the matrix and gain egress via the increased synthesis of proteolytic enzymes. Various investigations have shown altered regulation of the synthesis of ECM proteins in various malignant cells. The hypothesis of this proposal is that there is altered transcriptional regulation of ECM protein synthesis as a direct response to the transformed state, and that this alteration enhances metastasis. In this proposal, we propose a systematic approach to exploring this hypothesis. A battery of paired cell lines and tumor specimens (normal cells, polyps, primary tumors and metastatic samples) from breast, colon and skin will be analyzed by Northern, Southern and slot blot techniques for the increased/decreased transcription of genes encoding osteonectin, decorin, type I collagen, fibronectin, and various oncogenes. The housekeeping proteins, beta-actin and glycerol phosphate dehydrogenase, will serve as controls in these experiments. Nuclear extractions, DNA footprinting, gel shift mobility and site directed mutagenesis assays will be performed to determine the molecular mechanism(s) of the altered transcription. Data from these studies may provide a mechanism by which to determine the probability of metastasis at a specific stage of tumor growth, and provide molecular markers to determine if micro-metastasis has already occurred.