DEMONSTRATION OF CATHEPSIN FUNCTION IN TUMOR METASTASIS

描述：(改编自调查员的摘要)改变表达方式 组织蛋白酶B，而L与乳腺癌和许多其他人类 恶性肿瘤。这些改变涉及转录和转录的改变 细胞内的运输，使这些蛋白质的数量增加 靶向细胞表面或分泌到细胞外空间。这个 细胞外酶被认为促进了肿瘤的生长或转移， 可能是通过修饰细胞表面或细胞外基质 蛋白质。虽然组织蛋白酶与癌症之间的联系已为人所知 一段时间以来，人们对这些蛋白质如何参与 与癌症相关的过程。拟议研究的目标是提供 组织蛋白参与乳腺癌的直接证据 界定参与的性质，并开始探索 组织蛋白酶在细胞水平发挥作用。 拟议的研究将利用一组独特的小鼠乳腺肿瘤细胞 在经历不同阶段的过程中能力不同的线条 转移。在初步研究中，最高转移率的株系 SET被发现分泌升高的金属蛋白酶水平和 纤溶酶原激活剂，或溶酶体组织蛋白。有多种表型 观察到组织蛋白酶的分泌，表明有多种机制 牵涉其中。例如，其中一个细胞系(66c14)显示为泛化 分泌组织蛋白和其他溶酶体蛋白，而另一条线 (4T07)选择性分泌组织蛋白酶L。确定…的牵连 组织蛋白B、D和L在肿瘤进展中的作用组织蛋白会过度表达 或在这些细胞系中被消融，以及对体内生长和 转移情况将被确定。组织蛋白酶的过度表达将 用组织蛋白酶转染鼠乳腺肿瘤细胞系实现的 CDNA和组织蛋白酶的消融将通过基因打靶实现 方法论。分子和细胞生物学技术将用于 确定观察到的组织蛋白酶变化的分子基础 合成和贩运，并在细胞水平上进一步调查 组织蛋白如何参与癌症相关进展。从这些 通过研究，他们希望(I)更好地了解溶酶体是如何 蛋白水解酶参与癌症的进展，(Ii)一个理解 与癌症相关的细胞和分子机制 组织蛋白表达和贩运的变化和(3)信息 这可能有助于更有效地诊断和治疗 疾病。

DESCRIPTION: (adapted from the investigator's abstract) Altered expression of cathepsins B, and L is associated with breast cancer and many other human malignancies. These alterations involve changes in both transcription and intracellular trafficking such that increased amounts of these proteins are targeted to the cell surface or secreted into the extracellular space. The extracellular enzymes are thought to facilitate tumor growth or metastasis, perhaps through modification of cell surface or extracellular matrix proteins. While the association of cathepsins with cancer has been known for some time, little is known about how these proteins participate in cancer-related processes. The goals of the proposed research are to provide direct evidence for the involvement of cathepsins in breast cancer, to define the nature of the involvement, and to begin to explore the basis for cathepsin function at the cellular level. The proposed studies will utilize a unique set of mouse breast tumor cell lines which differ in their ability to progress through various stages of metastasis. In preliminary studies, the most highly metastatic lines of the set were found to secrete either elevated levels of metalloproteinases and plasminogen activators, or lysosomal cathepsins. Multiple phenotypes were observed for cathepsin secretion, indicating that multiple mechanisms are involved. For example, one of the cell lines (66c14) displayed generalized secretion of cathepsins and other lysosomal proteins, while another line (4T07) secreted cathepsin L selectively. To establish the involvement of cathepsins B, D and L in tumor progression, cathepsins will be overexpressed or ablated in these cell lines and the effects on in vivo growth and metastasis will be determined. Cathepsin overexpression will be accomplished by transfection of mouse breast tumor cell lines with cathepsin cDNAs, and cathepsin ablation will be achieved by gene targeting methodology. Molecular and cell biologic techniques will be used to determine the molecular basis for the observed alterations in cathepsin synthesis and trafficking and to further investigate at the cellular level how cathepsins participate in cancer-related progresses. From these studies, they hope to acquire (i) a better understanding of how lysosomal proteinases participate in the progression of cancer, (ii) an understanding of the cellular and molecular mechanisms responsible for cancer-related alterations in cathepsin expression and trafficking and (iii) information that may be of use for more effective diagnosis and treatment of the disease.