Effects of HAl-i andHAI-2 on the growth and invasion of human glioblastoma cells

HAl-i和HAI-2对人胶质母细胞瘤细胞生长和侵袭的影响

• 批准号: 13671449
• 负责人: NAKANO Shinichi
• 金额: $ 1.09万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671449/
• 关键词: Hepatocyte growth factor activator inhibitor (HAI); HAI-1; HAI-2; glioblastoma; Glioma; HGF; ヒト神経膠腫; HGFA; 腫瘍浸潤能抑制

Hepatocyte growth factor activator inhibitor (HAl) was initially identified as a potent endogenous inhibitor of hepatocyte growth factor activator (HGFA) that is responsible for the activation of hepatocyte growth factor/scatter factor. To date two kinds of HAI have been identified, namely HAI-1 and HAI-2. Both are Kunitz-type serine proteinase inhibitors and inhibit not only HGFA but also other serine proteinases such as plasmin and trypsin. Notably, each HAI has a transmembrane domain near the carboxyl terminal end, and thus, HAI is an integral-membrane proteinase inhibitor that must have important regulatory role on the cellular surface. To explore the possible role of HAI-1 and HAI-2 in invasive growth of tumor cells, we analyzed the effects of experimental over-expression of each HAI on glioblastoma (GBM), a malignant brain tumor that is highly invasive. Two cultured human GBM cell lines (U251 and YKG-1) were stably transfected with an expression vector harboring human HAI-1 or HAI-2 cDNA. Subsequent in vitro analyses indicated that the over-expression of HAI-2 significantly suppressed the fibrinolytic activity, as well as the invasion into the fibrin gel, of both GBM cell lines. On the other hand, the expression of HAI-1 showed much less effect relative to HAI-2 on the fibrinolytic activity of GBM cells. Then, we analyzed the effects of HAIs on in vivo invasive growth of these cell lines. The GBM cells were injected into the nude mice brain. Eight weeks after the injection, the mice were sacrificed and necropsied in order to determine the development of tumor. Rather surprisingly, HAI-2-transfected clones showed enhanced frequency of the tumor formation in vivo and formed larger tumor than control clones. These results indicated that HAI, particularly HAI-2, might have undetermined important function on the survival and/or growth of tumor cells in vivo, which is dependent or independent of its proteinase-inhibitory activity.

肝细胞生长因子激活物抑制物(Hal)最初被认为是肝细胞生长因子激活物(HGFA)的内源性抑制物，负责激活肝细胞生长因子/分散因子。到目前为止，已鉴定出两种HAI，即HAI-1和HAI-2。这两种都是库尼茨类型的丝氨酸酶抑制剂，不仅能抑制HGFA，还能抑制其他丝氨酸蛋白酶，如纤溶酶和胰蛋白酶。值得注意的是，每个HAI在羧基末端附近都有一个跨膜结构域，因此，HAI是一种完整的膜蛋白水解酶抑制剂，必须对细胞表面起到重要的调节作用。为了探讨HAI-1和HAI-2在肿瘤细胞侵袭性生长中的可能作用，我们分析了实验性过表达HAI-1和HAI-2对恶性脑肿瘤胶质母细胞瘤(GBM)的影响。将含有人HAI-1和HAI-2基因的真核表达载体分别稳定地导入人GBM细胞系U251和YKG-1。随后的体外分析表明，HAI-2的过表达显著抑制了两种GBM细胞的纤溶活性和对纤维蛋白凝胶的侵袭。另一方面，与HAI-2相比，HAI-1的表达对GBM细胞纤溶活性的影响要小得多。然后，我们分析了HAIs对这些细胞系体内侵袭性生长的影响。将GBM细胞注射到裸鼠脑内。注射8周后，处死小鼠，进行尸检，以确定肿瘤的发展情况。更令人惊讶的是，HAI-2转基因克隆在体内显示出更高的肿瘤形成频率，并形成了比对照克隆更大的肿瘤。这些结果表明，HAI，尤其是HAI-2，可能对体内肿瘤细胞的存活和/或生长具有未知的重要作用，这种作用依赖于或不依赖于其抑制蛋白水解酶的活性。

项目成果

濱砂亮一, 他: "Reduced expression of hepatocyte growth factor activator inhibitor type-2/placental bikunin(HAI-2/PB) in human glioblastomas : implication for anti-invasive role of HAI-2/PB in glioblastoma cells"Int J Cancer. 93(3). 339-345 (2001)

Ryoichi Hamasa 等人：“人胶质母细胞瘤中肝细胞生长因子激活剂抑制剂 2 型/胎盘 Bikunin (HAI-2/PB) 的表达降低：暗示 HAI-2/PB 在胶质母细胞瘤细胞中的抗侵袭作用”Int癌症杂志93（3）。

Hiroaki Kataoka, Hiroshi Itoh, Takeshi Shimomura, Yoshitsugu Nuki, Seiji Naganuma, Keiji Miyazawa: "Regulation of Hepatocyte Growth Factor (HGF) Activation on Cell Surface : Insights into an Emerging Class of Cell Surface Protease Inhibitors."LIfeXY. 1. 1

Hiroaki Kataoka、Hiroshi Itoh、Takeshi Shimomura、Yoshitsugu Nuki、Seiji Naganuma、Keiji Miyazawa：“细胞表面肝细胞生长因子 (HGF) 激活的调节：对新兴细胞表面蛋白酶抑制剂的见解。”LIfeXY。

Kataoka H, Itoh H, Nuki Y, Hamasuna R, Naganuma S, Kitamura N, Shimomura T: "Mouse Hepatocyte Growth Factor (HGF) Activator Inhibitor Type 2 Lacking the First Kunitz Domain Potently Inhibits the HG Activator."Biochem Biophys Res Commun. 290(3). 1096-1100

Kataoka H、Itoh H、Nuki Y、Hamasuna R、Naganuma S、Kitamura N、Shimomura T：“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 HG 激活剂。”Biochem Biophys Res Commun。

Itoh H, Kataoka H, Yamauchi M, Naganuma S, Akiyama Y, Nuki Y, Shimomura T, Miyazawa K, Kitamura N, Koono M: "Identification of hepatocyte growth factor activator inhibitor type 2 (HAI-2)-related small peptide (H2RSP) : its nuclear localization and generat

Itoh H、Kataoka H、Yamauchi M、Naganuma S、Akiyama Y、Nuki Y、Shimomura T、Miyazawa K、Kitamura N、Koono M：“肝细胞生长因子激活剂抑制剂 2 型 (HAI-2) 相关小肽的鉴定（