Roles of integral-membrane serine proteinase inhibitors in proliferation, migration and invasion of epithelial cells

整合膜丝氨酸蛋白酶抑制剂在上皮细胞增殖、迁移和侵袭中的作用

• 批准号: 12670209
• 负责人: KATAOKA Hiroaki
• 金额: $ 1.34万
• 依托单位: Miyazaki Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670209/
• 关键词: membrane-type protease inhibitor; Hepatocyte growth factor (HGF); HGF activator; HAI-1; amyloid beta protein precursor; colon cancer; colonic mucosa; knock-out mouse; 膜型プロテアーゼインヒビター; HAI-2

Cell surface proteolysis is an important mechanism for the generation of biologically active proteins that mediates a diverse range of cellular functions. Therefore the interactions between proteinases and cellular surface inhibitors must have important regulatory roles in cellular functions. There is a unique class of SPIs that is synthesized as a transmembrane glycoprotein. These SPIs have one or two extracellular Kunitz-type serine proteinase inhibitor domains, a presumed transmembrane domain near the carboxyl-terminus and a short intracytoplasmic domain. This unique group of SPIs consists of 4 distinct proteins at present. These are APP, APP-like protein 2 (APLP2), HAI-1 and HAI-2. In this project, the functions of these membrane-type inhibitors were extensively analyzed. Following are the results of the project (April, 2000 - March, 2002).(1) Roles of HAI-1 in the regulation of pericellular activation of HGFActivation of hepatocyte growth factor/scatter factor (HGF/SF) is a critic … More al limiting step in the HGF/SF-induced signaling pathway mediated by MET receptor tyrosine kinase. Although HGF/SF-MET signaling could have potentially important roles in the tissue regeneration and tumor progression, little is known about the regulation of HGF/SF activation. In this project, we have shown that HGF activator (HGFA) a recently identified factor XII-like serine proteinase, is critically involved in this process. Furthermore, we also showed that HGF activator inhibitor type 1 (HAI-1) should have an important regulatory role in the pericellular activation of HGF/SF having diverse roles acting as a cell surface specific inhibitor of active HGFA and a reservoir of this enzyme on the cell surface. The latter property might paradoxically ensure the concentrated pericellular HGFA activity in certain cellular conditions in which shedding of HAI-1/HGFA complex from the plasma membrane is upregulated.(2) APP is involved in the growth of colon carcinoma cells in vitro and in vivoIn a panel of human colon carcinoma cell lines, sAPP/PN-II was a major SPI secreted by all the cell lines examined. In this project we found that the downregulation of APP mRNA by an antisense mRNA strategy resulted in significantly reduced cellular proliferation of a colon carcinoma cell line in vitro, indicating that APP is somehow involved in the regulation of cellular proliferation. Of importance was the observation that reduced cellular growth was observed not only in vitro but also in vivo when the antisense colon carcinoma clones were transplanted into nude mice.(3) Generation of HGFA knock-out mouse and HAI-1 knock-out mouse.(4) Identification of a novel nuclear localization signal protein namely HAI-2-related small peptide (H2RSP). Less

细胞表面蛋白水解是产生生物活性蛋白的重要机制，其介导多种细胞功能。因此，蛋白酶和细胞表面抑制剂之间的相互作用必须在细胞功能中具有重要的调节作用。有一类独特的SPI是作为跨膜糖蛋白合成的。这些SPI具有一个或两个细胞外Kunitz型丝氨酸蛋白酶抑制剂结构域，羧基末端附近的假定跨膜结构域和短胞质内结构域。目前，这组独特的SPI由4种不同的蛋白质组成。这些是APP，APP样蛋白2（APLP 2），HAI-1和HAI-2。本项目对这些膜型抑制剂的功能进行了广泛的分析。以下是该项目的结果（2000年4月至2002年3月）。(1)HAI-1在肝细胞生长因子活化中的作用肝细胞生长因子/分散因子（HGF/SF）的活化是一个重要的研究领域 ...更多信息 MET受体酪氨酸激酶介导的HGF/SF诱导的信号通路中的一个限制步骤。尽管HGF/SF-MET信号在组织再生和肿瘤进展中可能具有潜在的重要作用，但对HGF/SF活化的调控知之甚少。在这个项目中，我们已经表明，肝细胞生长因子激活剂（HGFA），最近确定的因子XII样丝氨酸蛋白酶，是关键参与这一过程。此外，我们还表明，肝细胞生长因子激活抑制剂1型（HAI-1）应该有一个重要的调节作用，肝细胞生长因子/SF作为一个细胞表面特异性抑制剂的活性HGFA和水库的细胞表面上的这种酶具有不同的作用的细胞周围的激活。后一种性质可能矛盾地确保了在某些细胞条件下浓缩的细胞周HGFA活性，其中HAI-1/HGFA复合物从质膜的脱落被上调。(2)APP参与结肠癌细胞的体外和体内生长。在一组人结肠癌细胞系中，sAPP/PN-II是所有检测的细胞系分泌的主要SPI。在这个项目中，我们发现，APP mRNA的下调的反义mRNA的策略，导致显着降低细胞增殖的结肠癌细胞系在体外，表明APP在某种程度上参与调节细胞增殖。重要的是，当将反义结肠癌克隆移植到裸鼠中时，不仅在体外而且在体内观察到细胞生长减少。(3)HGFA敲除小鼠和HAI-1敲除小鼠的产生。(4)一种新的核定位信号蛋白HAI-2相关小肽（H2 RSP）的鉴定。少

项目成果

Meng JY, Kataoka H, Itoh H, Koono M:: "Amyloid β protein precursor is involved in the growth of human colon carcinoma cell in vitro and in vivo"International Journal of Cancer. 92. 31-39 (2001)

孟JY，Kataoka H，Itoh H，Koono M：：“β淀粉样蛋白前体参与体外和体内人类结肠癌细胞的生长”国际癌症杂志92. 31-39（2001）。

MENG, J-Y., KATAOKA, H., ITOH, H., KOONO, M.: "Amyloid β protein precursor is involved in the growth of human colon carcinoma cell in vitro and in vivo"Int. J. Cancer. 92. 31-39 (2001)

MENG, J-Y.、KATAOKA, H.、ITOH, H.、KOONO, M.：“β 淀粉样蛋白前体参与人结肠癌细胞的体外和体内生长”Int. 92。 31 -39 (2001)

Itoh H, Kataoka H, et al.: "Identification of hepatocyte growth factor activator inhibitor type 2 (HAI-2)-related small peptide (H2RSP): Its nuclear localization and generation of chimeric mRNA transcribed from both HAI-2 and H2RSP genes"Biochemical Bioph

Itoh H、Kataoka H 等人：“肝细胞生长因子激活剂抑制剂 2 型 (HAI-2) 相关小肽 (H2RSP) 的鉴定：其核定位以及从 HAI-2 和 H2RSP 基因转录的嵌合 mRNA 的生成

Kataoka H, Itoh H, et al.: "Mouse hepatocyte growth factor (HGF) activator inhibitor type 2 lacking the first Kunitz domain potently inhibits the HGF activator"Biochemical Biophysical Research Coummun.. 290. 1096-1100 (2002)

Kataoka H、Itoh H 等人：“缺少第一个 Kunitz 结构域的小鼠肝细胞生长因子 (HGF) 激活剂抑制剂 2 型可有效抑制 HGF 激活剂”Biochemical Biophysical Research Coummun.. 290. 1096-1100 (2002)

Kataoka H, Itoh H, Koono M :: "Emerging multifunctional aspects of cellular serine proteinase inhibtors in tumor progression and tissue regeneration"Pathlogy International. 52. 89-102 (2002)

Kataoka H、Itoh H、Koono M ::“细胞丝氨酸蛋白酶抑制剂在肿瘤进展和组织再生中的新兴多功能方面”国际病理学。