Evaluation of MMP inhibitors for cancer treatment

MMP抑制剂用于癌症治疗的评价

• 批准号: 10557016
• 负责人: SEIKI Motoharu
• 金额: $ 7.62万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557016/
• 关键词: MMP; MT-MMP; MMP inhibitor; Invasion and metastasis; マトリックスメタロプロテアーゼ; 膜型酵素; がんの浸潤・転移; MT1-MMP; 細胞外基質分解酵素

Matrix metalloproteinases (MMPs) degrade extracellular matrix (ECM) and support cancer cell invasion in tissue, and eventually metastasis formation. Inhibition of MMP activity is expected to prevent cancer cell growth, invasion and metastasis, and angiogenesis. Among the MMPs, membrane-type MMPs (MT-MMPs) are thought to be important for cancer cell invasiohbecause that are responsible for perice11ular degradation of ECM that is required for cells to traverse in tissue. So far five MT-MMPs are known. One of them, MT1-MMP, is frequently expressed in cancer cells and activates pro-gelatinase A which is able to degrade type IV collagen in the basement membrane.(1) Localization of MT1-MMP was found to be regulated by associating with actin cytoskelton within the cells.(2) To detect MT-MMP activities at subcellular level, we established an assay system by using either FITC-coated gelatin film or DQ-gelatin of which fuluorecense can be activated by proteolytic cleavage.(3) Preparation of catalitically active MT1, MT2, MT3, MT4 and MT5-MMPs were successful by using an expression system in E. coli.

基质金属蛋白酶（MMP）降解细胞外基质（ECM）并支持癌细胞在组织中的侵袭，并最终形成转移。MMP活性的抑制预期防止癌细胞生长、侵袭和转移以及血管生成。在MMPs中，膜型MMPs（membrane-type MMPs，MT-MMPs）被认为对癌细胞的侵袭是重要的，因为其负责细胞在组织中穿过所需的ECM的细胞外基质的降解。到目前为止，已知五种MT-MMP。其中之一，MT 1-MMP，经常在癌细胞中表达并激活明胶酶A，其能够降解基底膜中的IV型胶原。(1)MT 1-MMP的定位被发现是通过与细胞内的肌动蛋白结合来调节的。(2)为了在亚细胞水平上检测MT-MMP活性，我们建立了一个检测系统，通过使用FITC包被的明胶膜或DQ-明胶，其中fuluorecense可以被蛋白水解裂解激活。(3)利用大肠杆菌表达系统成功地制备了具有催化活性的MT 1、MT 2、MT3、MT4和MT5-MMPs。杆菌

项目成果

清木 元治: "Immunohistochemical study of MT-MMP tissue status in gastric carcinoma and correlation with survival analyzed by univariate and multivariate analysis." OncoL Rep.5. 1485-1488 (1998)

Motoharu Kiyoki：“胃癌 MT-MMP 组织状态的免疫组织化学研究以及通过单变量和多变量分析分析与生存的相关性。”OncoL Rep.5 (1998)。

Shofuda, K., K. Moriyama, A. Nishihashi, S. Higashi, H. Mizushima, H. Yasumitsu, K. Miki, H. Sato, M. Seiki, and K. Miyazaki: "Role of tissue inhibitor of metalloproteinases-2 (TIMP-2) in regulation of pro-gelatinase A activation by membrane-type matrix m

Shofuda，K.，K. Moriyama，A. Nishihashi，S. Higashi，H. Mizushima，H. Yasumitsu，K. Miki，H. Sato，M. Seiki 和 K. Miyazaki：“金属蛋白酶组织抑制剂的作用-

Shofuda, K. et al.: "Role of tissue inhibitor of metalloproteinase-2 in regulation of p@ro-gelatinase A activation catalyzed by membrane-type matrix metalloproteinase-1 in human cancer cells"J. Biochem.. 124. 462-470 (1998)

Shofuda, K. 等人：“金属蛋白酶 2 组织抑制剂在调节人类癌细胞中膜型基质金属蛋白酶 1 催化的 p@ro 明胶酶 A 激活中的作用”J.

Akizawa, T., T. Uratani, M. Matsukawa, A. Kunimatsu, Y. Ito, M. Itoh, Y. Ohshiba, M. Yamada, and M. Seiki: "Development and application of a microplate assay method for the muss screening of MMP inhibitors"Ann N Y Acad Sci. 878. 622-624 (1999)

Akizawa, T.、T. Uratani、M. Matsukawa、A. Kunimatsu、Y. Ito、M. Itoh、Y. Ohshiba、M. Yamada 和 M. Seiki：“贻贝微孔板测定方法的开发和应用

清木 元治: "Overexpression of tissue inhibitor of matrix metalloproteinase-1(TIMP-1) in metastatic MDCK cells transformed by v-src." Clin.Exp.Metastasis. (in press). (1999)

Motoharu Kiyoki：“在 v-src 转化的转移性 MDCK 细胞中过度表达基质金属蛋白酶-1 (TIMP-1)”（正在出版）。