Activation of MMP-9 by membrane type-1 MMP/MMP-2 axis stimulates tumor metastasis.
Activation of MMP-9 by membrane type-1 MMP/MMP-2 axis stimulates tumor metastasis.
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DOI：
10.1111/cas.13134
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发表时间：
2017-03
期刊：
影响因子：
5.7
通讯作者：
Sato H
中科院分区：
文献类型：
--
作者：
Li Z;Takino T;Endo Y;Sato H
通过将MMP-1的组织抑制剂（TIMP-1）与II型跨膜镶嵌丝氨酸蛋白酶（MSP-T1）融合来产生proMMP-9的人工受体。293 T细胞中MSP-T1的表达诱导proMMP-9的结合，proMMP-9被膜型1 MMP（MT 1-MMP）激活的MMP-2加工。用MSP-T1基因转染的HT 1080细胞在胶原凝胶中产生活化的MMP-9，并且向培养物中添加proMMP-2增强了它，这导致了强烈的胶原消化。这些细胞转移到鸡胚肝比对照细胞。在存在外源性proMMP-2的情况下，用伴刀豆球蛋白A处理HT 1080细胞不仅诱导proMMP-2的活化，还诱导proMMP-9的活化。用siRNA敲低MT 1-MMP或TIMP-2表达可抑制proMMP-2和proMMP-9的活化。TIMP-1 siRNA的转染抑制了proMMP-9的细胞结合和活化，但不抑制proMMP-2的活化。去整合素和金属蛋白酶10（ADAM 10）表达的敲低降低了proMMP-9的细胞结合和加工。这些结果表明，与含有TIMP-1和ADAM 10的受体复合物结合的proMMP-9被MT 1-MMP/MMP-2轴激活，因此激活的MMP-9刺激细胞蛋白水解和转移。
An artificial receptor for proMMP‐9 was created by fusing tissue inhibitor of MMP‐1 (TIMP‐1) with type II transmembrane mosaic serine protease (MSP‐T1). Expression of MSP‐T1 in 293T cells induced binding of proMMP‐9, which was processed by MMP‐2 activated by membrane type 1 MMP (MT1‐MMP). HT1080 cells transfected with the MSP‐T1 gene produced activated MMP‐9 in collagen gel, and addition of proMMP‐2 to the culture augmented it, which resulted in intensive collagen digestion. These cells metastasized into chick embryonic liver more than control cells. Treatment of HT1080 cells with concanavalin A in the presence of exogenous proMMP‐2 induced activation of not only proMMP‐2 but also proMMP‐9. Knockdown of MT1‐MMP or TIMP‐2 expression with siRNA suppressed activation of both proMMP‐2 and proMMP‐9. Transfection of TIMP‐1 siRNA suppressed cell binding and activation of proMMP‐9, but not proMMP‐2 activation. Knockdown of a disintegrin and metalloproteinase 10 (ADAM10) expression reduced cell binding and processing of proMMP‐9. These results suggest that proMMP‐9, which binds to a receptor complex containing TIMP‐1 and ADAM10, is activated by the MT1‐MMP/MMP‐2 axis, and MMP‐9 thus activated stimulates cellular proteolysis and metastasis.