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Metabolism of Proteoglycans in the Extracellular Matrix

细胞外基质中蛋白多糖的代谢

基本信息

批准号：
11307040
负责人：
YANAGISHITA Masaki
金额：
$ 22.65万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11307040/
关键词：
extracellular matrix Proteoglycan heparan sulfate degradation enzyme heparanase

项目摘要

The extracellular matrix is one of the major constituents of connective tissues and functions as a molecular complex providing proper environments necessary for cell development, differentiation and metabolism. Together with collagens, proteoglycans play major constitutive roles in the extracellular matrix. We have studied the metabolic regulation of extracellular matrix, especially focusing on the catabolism of heparan sulfate proteoglycans and a degrading enzyme, heparanase, necessary for this process. We have isolated a large quantity of the enzyme from cell cultures of rat parathyroid cell line (PTr) and characterized its enzymatic properties in detail. We have determined the nucleotide sequence of the mRNA encoding heparanase. We also investigated roles of extracellular matrix degradation by heparanase in the mechanism of cancer cell invasion and metastasis, using clinical specimens of lung cancers, oral squamous cell carcinomas and esophageal squamous cell carcinomas. The major results are summarized as follow.1. Heparanase, a specific enzyme which degrades heparan sulfate proteoglycans, was isolated and its enzymatic properties, including the substrate structural requirements were studied in detail.2. Nucleotide sequence of the gene encoding heparanase was determined.3. Monoclonal and polyclonal antibodies against heparanase have been prepared. Characterization of these antibodies is still under way by immunohistochemical and Western blot procedures.4. Good correlation between the invasiveness and the production of heparanase in lung cancers, oral squamous cell carcinomas and esophageal squamous cell carcinomas has been established. Studies on roles of heparanase in the mechanism of cancer invasion are under way.

细胞外基质是结缔组织的主要成分之一，并且作为分子复合物发挥功能，为细胞发育、分化和代谢提供必要的适当环境。与胶原蛋白一起，蛋白聚糖在细胞外基质中发挥主要的组成性作用。我们已经研究了细胞外基质的代谢调节，特别是集中在硫酸乙酰肝素蛋白多糖和降解酶，乙酰肝素酶，这一过程所必需的催化剂。我们已经从大鼠甲状旁腺细胞系（PTr）的细胞培养物中分离了大量的酶，并详细地表征了其酶学性质。我们已经确定了编码乙酰肝素酶的mRNA的核苷酸序列。我们还研究了乙酰肝素酶降解细胞外基质在癌细胞侵袭和转移机制中的作用，使用肺癌，口腔鳞状细胞癌和食管鳞状细胞癌的临床标本。主要研究结果如下：1.分离纯化了一种特异性的硫酸乙酰肝素蛋白多糖降解酶--乙酰肝素酶，并对其酶学性质进行了研究，包括底物结构要求.测定了乙酰肝素酶基因的核苷酸序列.制备了抗乙酰肝素酶的单克隆和多克隆抗体。这些抗体的特征仍在通过免疫组织化学和Western印迹程序进行中。在肺癌、口腔鳞状细胞癌和食管鳞状细胞癌中，乙酰肝素酶的产生与侵袭性之间已经建立了良好的相关性。对乙酰肝素酶在肿瘤侵袭机制中的作用的研究正在进行中。