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Activation of matrix metalloproteinases on cell surface

细胞表面基质金属蛋白酶的激活

基本信息

批准号：
10044244
负责人：
SEIKI Motoharu
金额：
$ 4.74万
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-10044244/
关键词：
Extracellular matrix Invasion and metastasis Matrix metalloproteinase Gelatinase A MT-MMP MMP MT-1-MMP ゼラチナーゼA がんの浸潤・転移

项目摘要

(1) Malignant melanoma cells degrade gelatin gel beneath the cells by extending protrusions called invadpodia. We foud that MT1-MMP localizes on the invadpodia. To monitor the cell surface. MT1-MMP more easily, we made MT1MMP fused with GFP and expressed it in the melanoma cells. Localization of GFP/MT1-MMP was clearly observed at invade podia where actin bundle can be seen. MT1-MMP was also detected along with actin stress fibers when it is expressed in on invasive CHO-K1 cells. Thus, localization of MT1-nMMP seems to be regulated through actin cytoskelton within the cells.(2) MT1-MMP activates pro-gelatinase Aby introducing cleavage in the propeptide domain in vitro. Expression of MT1-MMP in cultured cells induce cell-mediated activation of pro-gelatinase A. To confirm whether MT1-MMP can act as the activator of pro-gelatinase A in vivo, we generatedgene knockout mice by substituting the 1st to 4th exons of mouse genome to LacZ and Neo gene. Wild type mouse express MT1-MMP at high levels in various tissue during embryogenesis and activation of pro-gelatinase A in the tissue can be observed. In the knockout mice, however, such actibation of pro-gelatinase A could not be observed anymore. Thus, we conclued that MT1-MMP is the physiological activator of pro-gelatinase A.

(1)恶性黑色素瘤细胞通过延伸称为侵袭足的突起来降解细胞下的明胶凝胶。我们发现MT1-MMPs定位于入侵足纲。来监测细胞表面。MT1-MMPs更容易与GFP融合，并在黑色素瘤细胞中表达。GFP/MT1-MMP在侵袭性足部有明显的定位，可见肌动蛋白束。在侵袭性CHO-K1细胞上表达MT1-MMP时，它与肌动蛋白应激纤维一起被检测到。因此，MT1-NMMP的定位似乎是通过细胞内的肌动蛋白来调节的。(2)MT1-MMP在体外通过在前肽区域引入裂解来激活明胶酶原A。MT1-MMPs在培养细胞中的表达诱导明胶酶原A的激活。为了证实MT1-MMPs在体内是否可以作为明胶酶原A的激活剂，我们用LacZ和neo基因替换小鼠基因组的第1~4外显子，建立了基因敲除小鼠。在胚胎发育过程中，野生型小鼠在不同组织中高水平表达MT1-MMPs，并可观察到组织中明胶酶原A的激活。然而，在基因敲除的小鼠中，不能再观察到这种前明胶酶A的活性。因此，我们认为MT1-MMPs是明胶酶原A的生理激活剂。