Transmembrane/cytoplasmic domain-mediated membrane type 1-matrix metalloprotease docking to invadopodia is required for cell invasion
Transmembrane/cytoplasmic domain-mediated membrane type 1-matrix metalloprotease docking to invadopodia is required for cell invasion
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DOI：
10.1073/pnas.94.15.7959
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发表时间：
1997-07-22
影响因子：
11.1
通讯作者：
Chen, WT
中科院分区：
文献类型：
--
作者：
Nakahara, H;Howard, L;Chen, WT
人类恶性黑色素瘤细胞侵入细胞外基质（ECM）涉及在ECM降解位点的蛋白酶积累，其中基质金属蛋白酶（MMP）激活发生。这里，我们表明，当膜型1 MMP （MT-MMP）在RPMI7951人类黑色素瘤细胞中过表达时，细胞与ECM接触，激活可溶性和ECM结合的MMP-2，降解并侵入ECM。进一步的实验证明了MT-MMP定位对侵过性的重要性，未侵过性定位的MT-MMP过表达会激活可溶性MMP-2，但不会促进ECM降解或细胞侵袭性。用刀豆蛋白A上调内源性MT-MMP引起MMP-2的激活。然而，concanavin A的处理阻止了MT-MMP的侵入性定位和ECM的降解，缺乏跨膜（TM）和细胞质结构域的MT-MMP的截短突变体（Delta TMMT-MMP）和含有白细胞介素2受体α链（IL-2R） TM和细胞质结构域（Delta TMMT-MMP/TMIL-2R）的嵌合MT-MMP都没有定位于侵入性或ECM的降解。MT-MMP的TM/胞质结构域（TMMT-MMP）与mmp1的组织抑制剂（TIMP-1/TMMT-MMP）的嵌合体将TIMP-1分子引导到侵过体。因此，MT-MMP TM/细胞质结构域介导MT-MMP的空间组织进入侵殖体并随后降解ECM。
The invasion of human malignant melanoma cells into the extracellular matrix (ECM) involves the accumulation of proteases at sites of ECM degradation where activation of matrix metalloproteases (MMP) occurs, Here, we show that when membrane type 1 MMP (MT-MMP) was overexpressed in RPMI7951 human melanoma cells, the cells made contact with the ECM, activated soluble and ECM-bound MMP-2, and degraded and invaded the ECM, Further experiments demonstrated the importance of localization of the MT-MMP to invadopodia, Overexpression of MT-MMP without invadopodial localization caused activation of soluble MMP-2, but did not facilitate ECM degradation or cell invasiveness. Up-regulation of endogenous MT-MMP with concanavalin A caused activation of MMP-2. However, concanavalin A treatment prevented invadopodial localization of MT-MMP and ECM degradation, Neither a truncated MT-MMP mutant lacking transmembrane (TM) and cytoplasmic domains (Delta TMMT-MMP), nor a chimeric MT-MMP containing the interleukin 2 receptor alpha chain (IL-2R) TM and cytoplasmic domains (Delta TMMT-MMP/TMIL-2R) were localized to invadopodia or exhibited ECM degradation, Furthermore, a chimera of the TM/cytoplasmic domain of MT-MMP (TMMT-MMP) with tissue inhibitor of MMP 1 (TIMP-1/TMMT-MMP) directed the TIMP-1 molecule to invadopodia. Thus, the MT-MMP TM/cytoplasmic domain mediates the spatial organization of MT-MMP into invadopodia and subsequent degradation of the ECM.