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: 2683591
• 负责人: G. Gary Sahagian
• 金额: $ 27.02万
• 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/2683591
• 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：（改编自研究者的摘要）改变的表达 组织蛋白酶 B 和 L 与乳腺癌和许多其他人类癌症有关 恶性肿瘤。 这些改变涉及转录和 细胞内运输使得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 细胞外酶被认为促进肿瘤生长或转移， 也许通过细胞表面或细胞外基质的修饰 蛋白质。 虽然组织蛋白酶与癌症的关联已为人所知 一段时间以来，人们对这些蛋白质如何参与 癌症相关过程。 拟议研究的目标是提供 组织蛋白酶参与乳腺癌的直接证据 定义参与的性质，并开始探索参与的基础 组织蛋白酶在细胞水平上发挥作用。 拟议的研究将利用一组独特的小鼠乳腺肿瘤细胞 不同阶段的生产线的进展能力不同 转移。 在初步研究中，转移性最高的细胞系 发现一组分泌升高水平的金属蛋白酶和 纤溶酶原激活剂或溶酶体组织蛋白酶。 多种表型是 观察到组织蛋白酶的分泌，表明有多种机制 涉及。 例如，其中一个细胞系 (66c14) 显示为广义 组织蛋白酶和其他溶酶体蛋白的分泌，而另一条线 (4T07)选择性分泌组织蛋白酶L。 建立参与 组织蛋白酶B、D和L在肿瘤进展过程中，组织蛋白酶会过度表达 或在这些细胞系中被消除以及对体内生长和 将确定转移。 组织蛋白酶过度表达将 通过用组织蛋白酶转染小鼠乳腺肿瘤细胞系来完成 cDNA和组织蛋白酶消融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的进展。 从这些 通过研究，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) 了解 负责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以及 (iii) 信息 可能有助于更有效地诊断和治疗 疾病。