DEMONSTRATION OF CATHEPSIN FUNCTION IN TUMOR METASTASIS

组织蛋白酶在肿瘤转移中的功能演示

• 批准号: 2008765
• 负责人: G. Gary Sahagian
• 金额: $ 27.93万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2008765
• 关键词: breast neoplasms; carbohydrate receptor; cathepsin B; cathepsin D; cysteine endopeptidases; enzyme activity; enzyme induction /repression; gene mutation; intracellular transport; laboratory mouse; lysosomes; mannose 6 phosphate; metastasis; neoplasm /cancer genetics; protein biosynthesis; protein transport; tissue /cell culture

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.

描述：（改编自研究者摘要）表达改变 组织蛋白酶B和L与乳腺癌和许多其他人类疾病有关。 恶性肿瘤 这些改变涉及转录和转录水平的变化。 细胞内运输，使得这些蛋白质的量增加， 靶向细胞表面或分泌到细胞外空间。 的 胞外酶被认为促进肿瘤生长或转移， 可能通过修饰细胞表面或细胞外基质 proteins. 虽然已知组织蛋白酶与癌症的关联 一段时间以来，人们对这些蛋白质如何参与 癌症相关的过程。 本研究的目的是提供 组织蛋白酶参与乳腺癌的直接证据， 界定参与的性质，并开始探讨 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， 不同的路线，在他们的能力，通过不同阶段的进展， 转移 在初步研究中，最高转移性的品系， 组被发现分泌金属蛋白酶水平升高， 纤溶酶原激活剂或溶酶体组织蛋白酶。 多种表型 观察到组织蛋白酶分泌，表明多种机制， 涉案 例如，其中一个细胞系（66c14）显示全身性的 分泌组织蛋白酶和其他溶酶体蛋白，而另一条线 （4T07）选择性分泌组织蛋白酶L。 为了证实 组织蛋白酶B、D和L在肿瘤进展中， 或在这些细胞系中消融，以及对体内生长的影响， 将确定转移。 组织蛋白酶过表达将是 通过用组织蛋白酶转染小鼠乳腺肿瘤细胞系完成 cDNAs和组织蛋白酶切除将通过基因靶向来实现 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 合成和运输，并在细胞水平上进一步研究 组织蛋白酶如何参与癌症相关进展。 从这些 研究，他们希望获得（i）更好地了解溶酶体如何 蛋白酶参与癌症的进展，（ii）理解 癌症相关的细胞和分子机制 组织蛋白酶表达和运输的改变和（iii）信息 这可能有助于更有效地诊断和治疗 疾病