MMP-9 and -12 cause N-cadherin shedding and thereby β-catenin signalling and vascular smooth muscle cell proliferation
MMP-9 and -12 cause N-cadherin shedding and thereby β-catenin signalling and vascular smooth muscle cell proliferation
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DOI：
10.1093/cvr/cvn278
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发表时间：
2009-01-01
影响因子：
10.8
通讯作者：
George, Sarah J.
中科院分区：
文献类型：
--
作者：
Dwivedi, Amrita;Slater, Sadie C.;George, Sarah J.
血管平滑肌细胞（VSMC）增殖有助于再狭窄和动脉粥样硬化中的内膜增厚。以前，我们证明，基质降解金属蛋白酶（MMP）依赖脱落的细胞外部分的N-钙粘蛋白增加VSMC增殖通过升高β-连环蛋白信号和细胞周期蛋白D1的表达。在这项研究中，我们的目的是确定MMP-2，-9，-12，MMP-9或-14通过N-钙粘蛋白脱落调节VSMC增殖，MMP-9缺陷的增殖小鼠主动脉VSMC的N-钙粘蛋白脱落明显受损MMP-9（-/-））和MMP-12（MMP-12（-/-））与野生型对照相比（1.1 +/-0.7-和1.0 +/-0.1-对比2.0 +/-0.2-倍）。此外，与对照组相比，MMP-9或-12 siRNA敲低或MMP-9或-12缺陷的增殖VSMC显示出显著增加的N-钙粘蛋白细胞水平（分别为1.7 +/-0.2-、2.7 +/-0.6-和3.5 +/-1.6-、1.7 +/-0.2-倍）。与活性MMP-9或-12孵育的VSMC独立地增加N-钙粘蛋白裂解。此外，在MMP-9（-/-）和-12（-/-）增殖的VSMC中，β-连环蛋白信号传导分别显著降低了52 +/-17和81 +/-12%，这一点通过MMP-9和-12的siRNA敲低得到证实。MMP-9（-/-）和MMP-12（-/-）细胞中β-连环蛋白信号的降低与增殖和细胞周期蛋白D1蛋白水平的显著降低相一致。在所有实验中，用MMP-9和-12的组合调节观察到很少或没有累加效应。MMP-2和-14对N-cadherin脱落和VSMC增殖无明显影响。结论：MMP-9和-12促进内膜增厚是通过N-cadherin的独立切割，而N-cadherin的切割通过β-catenin信号通路促进VSMC增殖。
Vascular smooth muscle cell (VSMC) proliferation contributes to intimal thickening in restenosis and atherosclerosis. Previously, we demonstrated that matrix-degrading metalloproteinase (MMP)-dependent shedding of the extracellular portion of N-cadherin increased VSMC proliferation via elevation of beta-catenin signalling and cyclin D1 expression. In this study, we aimed to determine whether MMP-2, -9, -12, or -14 regulates VSMC proliferation via N-cadherin shedding.N-cadherin shedding was significantly impaired in proliferating mouse aortic VSMCs deficient in MMP-9 (MMP-9(-/-)) and MMP-12 (MMP-12(-/-)) compared with wild-type controls (1.1 +/- 0.7- and 1.0 +/- 0.1- vs. 2.0 +/- 0.2-fold). Furthermore, proliferating VSMCs subjected to MMP-9 or -12 siRNA knockdown or deficient in MMP-9 or -12 showed significantly increased cellular levels of N-cadherin compared with controls (1.7 +/- 0.2-, 2.7 +/- 0.6-, and 3.5 +/- 1.6-, 1.7 +/- 0.2-fold, respectively). Incubation of VSMCs with active MMP-9 or -12 independently increased N-cadherin cleavage. Additionally, beta-catenin signalling was significantly reduced by 52 +/- 17 and 81 +/- 12% in MMP-9(-/-) and -12(-/-) proliferating VSMCs, respectively, and this was corroborated by siRNA knockdown of MMP-9 and -12. Decreased beta-catenin signalling coincided with significantly reduced proliferation and cyclin D1 protein levels in MMP-9(-/-) and -12(-/-) cells. Little or no additive effect was observed with combined modulation of MMP-9 and -12 in all experiments. In contrast, N-cadherin shedding and VSMC proliferation were not modulated by MMP-2 and -14.In conclusion, we propose that MMP-9 and -12 promote intimal thickening by independent cleavage of N-cadherin, which elevates VSMC proliferation via beta-catenin signalling.