REGULATION OF ECM PROTEIN BIOSYNTHESIS IN NORMAL, MALIGNANT AND METASTATIC CELLS

正常细胞向恶性肿瘤的转化是一个多步骤的过程 由于基因改变和环境侮辱而产生的过程。 大多数恶性细胞都有能力通过 细胞脱离原发肿瘤，向内、向外渗出 在远端器官中建立继发性病灶。胞外 基质(ECM)和基底膜由各种 包裹在组织和器官周围并呈现不透性的蛋白质 活跃细胞移动的障碍。转移过程包括 游走性恶性细胞与肿瘤大分子的相互作用 基底膜和细胞外基质，细胞由此附着 通过增加合成增加到基质并获得出口 蛋白水解酶。各种调查显示， 多种恶性细胞中细胞外基质蛋白合成的调控。 这个提议的假设是，改变的转录 细胞外基质蛋白质合成的调节直接或间接相关 转化后的状态，进而增强转移能力。在这 建议，我们提出了一个系统的方法来探索这一假说。 一组配对的细胞系和肿瘤标本(正常细胞，息肉， 来自不同器官的原发肿瘤和转移样本)将 通过Northern、Southern和Slot印迹技术分析 增加/减少编码骨连蛋白、核心蛋白聚糖、 I型胶原、纤维连接蛋白和各种癌基因。家政服务 蛋白质，B-肌动蛋白，将在这些实验中作为对照。核子 提取、DNA足迹、凝胶移位和定点定向 将进行突变分析以确定分子 改变转录的机制(S)。来自这些研究的数据可能 提供一种确定转移概率的机制 在肿瘤生长的特定阶段，并提供分子标记以 确定微转移是否已经发生。

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 macromolecule 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 altered transcriptional regulation of ECM protein synthesis is directly or indirectly correlated with the transformed state, which in turn, 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 various organs 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 protein, B-actin, will serve as a control 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.