Membrane type-1 matrix metalloproteinase stimulates tumour cell-induced platelet aggregation: role of receptor glycoproteins
Membrane type-1 matrix metalloproteinase stimulates tumour cell-induced platelet aggregation: role of receptor glycoproteins
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DOI：
10.1038/sj.bjp.0705606
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发表时间：
2004-01-01
影响因子：
7.3
通讯作者：
Radomski, MW
中科院分区：
文献类型：
--
作者：
Alonso-Escolano, D;Strongin, AY;Radomski, MW
1基质金属蛋白酶-2（MMP-2）在激动剂和肿瘤细胞诱导的血小板聚集（TCIPA）中发挥作用。2 MMP-2作为酶原合成，并在细胞表面被膜型1基质金属蛋白酶激活（MT1-MMP，MMP-14）。3肿瘤细胞相关的MT 1-使用稳定共表达整合素α v β 3与MT 1-MMP的人乳腺癌MCF 7细胞研究TCIPA的MMP，单独表达α v β 3的细胞和模拟转染的细胞。4蛋白质印迹和酶谱证实α v β 3/MT 1-MMP细胞表达MT 1-MMP并有效地将proMMP-2加工成MMP-2。5聚集测定法，使用相差显微镜、透射电子显微镜和流式细胞术表征由MCF 7细胞系诱导的TCIPA。6细胞的聚集能力为：α v β 3/MT 1-MMP > α v β 3 =模拟细胞，如聚集测定法和相差显微镜所示。7电子显微镜显示，TCIPA过程中活化血小板和α v β 3/MT 1-MMP细胞之间的膜-膜相互作用。8用中和抗MMP-2抗体抑制MMP-2（5 μ M ml（-1））和邻菲咯啉（100 μ M）可减少α v β 3/MT 1-MMP细胞诱导的聚集。α v β 3/MT 1-MMP细胞诱导的TCIPA也可通过抑制腺苷三磷酸双磷酸酶的生成和作用而减少（250 μ g/ml）和2-甲硫基-AMP（2-MeSAMP）（30 μ M），但不是N-6-甲基-2 '-脱氧腺苷-3'，5 '-二磷酸盐（MRS 2179）（30 μ M）。10流式细胞术表明，TCIPA增强了糖蛋白（GP）Ib和IIb的表达。IIIa受体不仅在血小板上，而且在乳腺癌细胞上。11因此，（a）人乳腺癌细胞表面相关的MT 1-MMP，通过激活proMMP-2，刺激TCIPA;（B）ADP通过刺激P2 Y（12）受体放大MMP的作用，以及（c）肿瘤和血小板来源的GPIb和GPIIb/GPIIb两者。IIIa参与MT 1-MMP的聚集作用。
1 Matrix metalloproteinase-2 (MMP-2) plays a role in agonist- and tumour cell-induced platelet aggregation (TCIPA).2 MMP-2 is synthesized as a proenzyme and is activated at the cell surface by membrane type-1 matrix metalloproteinase (MT1-MMP, MMP-14).3 The significance of tumour cell-associated MT1-MMP for TCIPA was investigated using human breast carcinoma MCF7 cells stably coexpressing the integrin alphavbeta3 with MT1-MMP, cells expressing alphavbeta3 alone and mock-transfected cells.4 Western blot and zymography confirmed that alphavbeta3/MT1-MMP cells expressed MT1-MMP and efficiently processed proMMP-2 to MMP-2.5 Aggregometry, phase-contrast and transmission electron microscopy and flow cytometry were used to characterize TCIPA induced by MCF7 cell lines.6 The aggregating potency of cells was: alphavbeta3/MT1-MMP >alphavbeta3=mock cells, as shown by aggregometry and phase-contrast microscopy.7 Electron microscopy revealed close, membrane-membrane interactions between activated platelets and alphavbeta3/MT1-MMP cells during TCIPA.8 Inhibition of MMP-2 with the neutralizing anti-MMP-2 antibody (5 mug ml(-1)) and o-phenanthroline (100 muM) reduced aggregation induced by alphavbeta3/MT1-MMP cells.9 TCIPA induced by alphavbeta3/MT1-MMP cells was also reduced by inhibiting the generation and actions of ADP with apyrase (250 mug ml(-1)) and 2-methylthio-AMP (2-MeSAMP) (30 muM), but not N-6-methyl-2'-deoxyadenosine-3',5'-bisphosphate (MRS2179) (30 muM).10 Flow cytometry demonstrated that TCIPA enhanced expression of glycoprotein (GP) Ib and IIb/IIIa receptors not only on platelets but also on breast cancer cells.11 Thus, (a) human breast carcinoma cell surface-associated MT1-MMP, via activating proMMP-2, stimulates TCIPA; (b) ADP amplifies the effects of MMPs via stimulation of P2Y(12) receptors and (c) both tumour- and platelet-derived GPIb and GPIIb/IIIa are involved in the aggregatory effects of MT1-MMP.