Takino Takahisa, Koshikawa Naohiko, Miyamori Hisashi, Tanaka Motohiro, Sasaki Takuma, Okada Yasunori, Seiki Motoharu, Sato Hiroshi
Department of Molecular Virology and Oncology, Cancer Research Institute, Kanazawa University, Kanazawa 920-0934, Japan.
Oncogene. 2003 Jul 24;22(30):4617-26. doi: 10.1038/sj.onc.1206542.
A human placenta cDNA library was screened by the expression cloning method for gene products that interact with matrix metalloproteinases (MMPs), and we isolated a cDNA whose product formed a stable complex with pro-MMP-2 and pro-MMP-9. The cDNA encoded the metastasis suppressor gene KiSS-1. KiSS-1 protein was shown to form a complex with pro-MMP. KiSS-1 protein is known to be processed to peptide ligand of a G-protein-coupled receptor (hOT7T175) named metastin, and suppresses metastasis of tumors expressing the receptor. Active MMP-2, MMP-9, MT1-MMP, MT3-MMP and MT5-MMP cleaved the Gly118-Leu119 peptide bond of not only full-length KiSS-1 protein but also metastin decapeptide. Metastin decapeptide induced formation of focal adhesion and actin stress fibers in cells expressing the receptor, and digestion of metastin decapeptide by MMP abolished its ligand activity. Migration of HT1080 cells expressing hOT7T175 that harbor a high-level MMP activity was only slightly suppressed by either metastin decapeptide or MMP inhibitor BB-94 alone, but the combination of metastin decapeptide and BB-94 showed a synergistic effect in blocking cell migration. We propose that metastin could be used as an antimetastatic agent in combination with MMP inhibitor, or MMP-resistant forms of metastin could be developed and may also be efficacious.
通过表达克隆法筛选人胎盘cDNA文库，以寻找与基质金属蛋白酶（MMPs）相互作用的基因产物，我们分离出一个cDNA，其产物与前MMP-2和前MMP-9形成稳定复合物。该cDNA编码转移抑制基因KiSS-1。KiSS-1蛋白被证明与前MMP形成复合物。已知KiSS-1蛋白可被加工成一种名为metastin的G蛋白偶联受体（hOT7T175）的肽配体，并抑制表达该受体的肿瘤的转移。活性MMP-2、MMP-9、MT1-MMP、MT3-MMP和MT5-MMP不仅切割全长KiSS-1蛋白的Gly118-Leu119肽键，也切割metastin十肽。Metastin十肽在表达该受体的细胞中诱导粘着斑和肌动蛋白应力纤维的形成，MMP对metastin十肽的消化消除了其配体活性。单独使用metastin十肽或MMP抑制剂BB-94对具有高水平MMP活性的表达hOT7T175的HT1080细胞迁移的抑制作用仅略有抑制，但metastin十肽和BB-94的组合在阻断细胞迁移方面显示出协同作用。我们提出，metastin可与MMP抑制剂联合用作抗转移剂，或者可以开发MMP抗性形式的metastin，其可能也有效。
