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

已知肿瘤转移是由于金属蛋白水解酶升高所致。 癌细胞产生/活动导致破坏 细胞外基质(ECM)和基底膜，作为屏障 转移细胞通过循环和循环的进出 淋巴系统。几乎没有进行过研究来确定监管 结缔组织蛋白(CTP)在肿瘤中的合成状况 进步。这项研究的目标是确定其他 调控事件发生在癌细胞中，这进一步促进了 转移过程。具体地说，本研究将探索 细胞外基质蛋白质合成下调是一个整体的可能性 基底膜结构完整性丧失的成分和 ECM，从而促进转移。我们之前的研究表明 核心蛋白聚糖、纤维连接蛋白、骨连接蛋白和I型蛋白表达的改变 比较不同乳房结肠组织转录谱中的胶原蛋白 和皮肤(正常、良性、原发癌症和转移)细胞株和 临床样本。此外，限制片段长度多态 (RFLP)分析揭示了突变的基因(核心蛋白， 纤维连接蛋白和骨连接蛋白)在两种转移细胞系(黑色素瘤和骨连接蛋白)中的表达 乳房)。这项建议的具体目标是：(1)进行系统的 正常、良性、原发癌和转移细胞系的分析 和临床样本进一步研究细胞外基质合成减少的相关性 和增加的转移潜能，(2)发展转移概况 细胞外基质产生减少，金属蛋白酶活性增加 临床来源的组织，(3)确定结构和功能 具有RFLP多态性的核心蛋白聚糖和骨连接蛋白结构域 通过单链构象多态(SSCP)分析，RT-PCR 克隆和序列分析以及(4)确定其调节区 精选的ECM基因及其结合蛋白参与了 观察到了变化的规则。上述具体目标的完成应 提供了一种确定原发肿瘤转移能力的方法 原位肿瘤。

It is known that metastasis results from increased metalloprotease production/activity by cancerous cells resulting in the breaching of the extracellular matrix (ECM) and basement membranes, which serve as barriers to ingress and egress of metastatic cells through the circulatory and lymphatic systems. Few studies have been done to ascertain the regulatory and synthetic status of the connective tissue proteins (CTP) during tumor progression. The goal of this research is to determine what other regulatory events occur in cancerous cells which further facilitate the process of metastasis. Specifically, this research will explore the possibility that down-regulation of ECM protein synthesis is an integral component in the loss of structural integrity of the basement membrane and ECM, thus facilitating metastasis. Our previous studies demonstrated altered expression of decorin, fibronectin, osteonectin and type I collagen when transcriptional profiles were compared among breast colon and skin (normal, benign, primary cancerous and metastatic) cell lines and clinical samples. Additionally, restriction fragment length polymorphism (RFLP) analyses have revealed mutationally altered genes (decorin, fibronectin and osteonectin) in two metastatic cell lines (melanoma and breast). The specific aims of this proposal are: (1) perform a systematic analysis of normal, benign, primary cancerous and metastatic cell lines and clinical samples for further correlations of decreased ECM synthesis and increased metastatic potential, (2) develop a metastatic profile of decreased ECM production and increased metalloprotease activity in clinical derived tissues, (3) determine the structural and functional domains of decorin and osteonectin which demonstrate RFLP polymorphisms via single strand conformational polymorphism (SSCP) analysis, RT-PCR cloning and sequence analysis and (4) determine regulatory regions of selected ECM genes and their binding proteins which are involved in the observed altered regulation. Completion of the above specific aims should provide a means of determining the metastatic capability of a primary tumor in situ.