CD9 participates in endothelial cell migration during in vitro wound repair
CD9 participates in endothelial cell migration during in vitro wound repair
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DOI：
10.1161/01.atv.20.2.360
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发表时间：
2000-02-01
影响因子：
8.7
通讯作者：
Lanza, F
中科院分区：
文献类型：
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作者：
Klein-Soyer, C;Azorsa, DO;Lanza, F
CD 9是一种广泛表达的四跨膜蛋白家族的膜蛋白，具有多种功能，如信号转导、细胞粘附和细胞运动。我们检测了抗CD 9单克隆抗体（ALMA.1）在模拟血管生成过程的体外机械创伤修复过程中对人血管内皮细胞（EC）迁移和增殖的影响。ALMA.1诱导伤口修复的剂量依赖性抑制，在20 μ g/mL时降低35 +/- 1.5%。只有细胞迁移受到影响，因为在病变边缘的内皮细胞的增殖率没有修改，因为抑制修复也观察到非增殖照射的内皮细胞。抗CD 63四跨膜蛋白（H5 C6）和对照小鼠IgG（MOPC-21）的单克隆抗体无活性。CD 9是EC表面最丰富的蛋白质之一，在用ALMA.1和抗β（1）（4 B4）或抗β（3）（SDF. 3）单克隆抗体进行的双标记免疫荧光实验中，CD 9与EC膜上的β（1）或β（3）整联蛋白共定位。此外，ALMA.1和4 B4对损伤修复具有累加抑制作用，而4 B4单独也抑制EC增殖。在跨膜Boyden型测定中，ALMA.1诱导了EC向纤连蛋白和玻连蛋白迁移的剂量依赖性抑制，在100 μ g/mL时抑制率分别为45+/-6%和31+/-10%。4 B4在10 μ g/mL时抑制向纤连蛋白的迁移，但对玻连蛋白没有影响。EC与固定化抗CD 9单克隆抗体的粘附诱导酪氨酸磷酸化蛋白水平，类似于与β（1）或β（3）整联蛋白相互作用期间观察到的水平。这些结果表明，在损伤修复和血管生成过程中，CD 9可能通过与整合素的合作参与EC粘附和迁移。因此，CD 9是在转移和动脉粥样硬化过程中抑制血管生成的潜在靶标。
CD9, a widely expressed membrane protein of the tetraspanin family, has been implicated in diverse functions, such as signal transduction, cell adhesion, and cell motility, We tested the effects of an anti-CD9 monoclonal antibody (ALMA.1) on the migration and proliferation of human vascular endothelial cells (ECs) during repair of an in vitro mechanical wound mimicking angiogenic processes. ALMA.1 induced dose-dependent inhibition of wound repair with a 35 +/- 1.5% decrease at 20 mu g/mL. Only cell migration was affected, because the rate of proliferation of ECs at the lesion margin was not modified and because the inhibition of repair was also observed for nonproliferating irradiated ECs. Monoclonal antibodies against CD63 tetraspanin (H5C6) and control mouse IgG (MOPC-21) were inactive. CD9, one of the most abundant proteins at the surface of ECs, colocalized with beta(1) or beta(3) integrins on EC membranes in double-labeling immunofluorescence experiments with ALMA.1 and an anti-beta(1) (4B4) or anti-beta(3) (SDF.3) monoclonal antibody. Moreover, ALMA.1 and 4B4 had additive inhibitory effects on lesion repair, whereas 4B4 alone also inhibited EC proliferation In transmembrane Boyden-type assays, ALMA.1 induced dose-dependent inhibition of EC migration toward fibronectin and vitronectin with 45+/-6% and 31+/-10% inhibition, respectively, at 100 mu g/mL. 4B4 inhibited migration toward fibronectin at 10 mu g/mL but had no effect in the case of vitronectin, Adhesion of ECs to immobilized anti-CD9 monoclonal antibodies induced tyrosine-phosphorylated protein levels similar to those observed during interactions with beta(1) or beta(3) integrins. These results point to the involvement of CD9 in EC adhesion and migration during lesion repair and angiogenesis, probably through cooperation with integrins. As such, CD9 is a potential target to inhibit angiogenesis in metastatic and atherosclerotic processes.