Intimal smooth muscle cells of porcine and human coronary artery express S100A4, a marker of the rhomboid phenotype in vitro
Intimal smooth muscle cells of porcine and human coronary artery express S100A4, a marker of the rhomboid phenotype in vitro
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DOI：
10.1161/01.res.0000262654.84810.6c
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发表时间：
2007-04-13
影响因子：
20.1
通讯作者：
Bochaton-Piallat, Marie-Luce
中科院分区：
文献类型：
--
作者：
Brisset, Anne C.;Hao, Hiroyuki;Bochaton-Piallat, Marie-Luce
我们报道了从正常猪冠状动脉中膜分离的平滑肌细胞（SMC）群体表现出不同的表型：梭形（S）和菱形（R）。与中膜相比，R-SMC从支架诱导的内膜增厚中恢复的比例更高，表明它们参与内膜增厚形成。我们的目的是在体外鉴定R-SMCs的标志物，并探索其在体内的可能表达。S-和R-SMC蛋白提取物进行了比较，通过2维聚丙烯酰胺凝胶电泳，然后串联质谱。发现S100 A4主要在R-SMC提取物中表达。使用单克隆S100 A4抗体，我们证实了S100 A4在R-SMC中高度表达，而在S-SMC中几乎检测不到。S100 A4与α-平滑肌肌动蛋白共定位于几个静止细胞的应力纤维中，并在迁移过程中上调。PDGF-BB、FGF-2或与内皮细胞共培养，将S-SMC调节为R-表型，增加了S100 A4在S-和R-SMC中的表达。R-SMC中S100 A4 mRNA的沉默降低了细胞增殖，表明该蛋白具有功能作用。S100 A4在正常猪冠状动脉中膜中不表达，但在支架诱导的内膜增厚的SMC中高度表达。在人类中，S100 A4在冠状动脉中膜中几乎检测不到，而在动脉粥样硬化和再狭窄冠状动脉病变的SMC中显著表达。我们的研究结果表明，S100 A4是猪R-SMCs在体外和内膜平滑肌细胞在内膜增厚的发展过程中的标志物。它也是大量人类动脉粥样硬化和再狭窄SMC的标志物。阐明S100 A4的功能可能有助于了解动脉粥样硬化和再狭窄过程的演变。
We reported that smooth muscle cell (SMC) populations isolated from normal porcine coronary artery media exhibit distinct phenotypes: spindle-shaped (S) and rhomboid (R). R-SMCs are recovered in higher proportion from stent-induced intimal thickening compared with media suggesting that they participate in intimal thickening formation. Our aim was to identify a marker of R-SMCs in vitro and to explore its possible expression in vivo. S-and R-SMC protein extracts were compared by means of 2-dimensional polyacrylamide gel electrophoresis followed by tandem mass spectrometry. S100A4 was found to be predominantly expressed in R-SMC extracts. Using a monoclonal S100A4 antibody we confirmed that S100A4 is highly expressed by R-SMCs and hardly detectable in S-SMCs. S100A4 was colocalized with alpha-smooth muscle actin in stress fibers of several quiescent cells and upregulated during migration. PDGF-BB, FGF-2 or coculture with endothelial cells, which modulate S-SMCs to a R-phenotype, increased S100A4 expression in both S- and R-SMCs. Silencing of S100A4 mRNA in R-SMCs decreased cell proliferation, suggesting a functional role for this protein. In vivo S100A4 was absent in normal porcine coronary artery media, but highly expressed by SMCs of stent-induced intimal thickening. In humans, S100A4 was barely detectable in coronary artery media and markedly expressed in SMCs of atheromatous and restenotic coronary artery lesions. Our results indicate that S100A4 is a marker of porcine R-SMCs in vitro and of intimal SMCs during intimal thickening development. It is also a marker of a large population of human atheromatous and restenotic SMCs. Clarifying S100A4 function might be useful to understand the evolution of atherosclerotic and restenotic processes.