MARCKS silencing differentially affects human vascular smooth muscle and endothelial cell phenotypes to inhibit neointimal hyperplasia in saphenous vein
MARCKS silencing differentially affects human vascular smooth muscle and endothelial cell phenotypes to inhibit neointimal hyperplasia in saphenous vein
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DOI：
10.1096/fj.08-114173
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发表时间：
2009-02-01
期刊：
影响因子：
4.8
通讯作者：
LoGerfo, Frank W.
中科院分区：
文献类型：
--
作者：
Monahan, Thomas S.;Andersen, Nicholas D.;LoGerfo, Frank W.
内膜增生（IH）限制了所有心血管静脉旁路移植物的通畅性。我们以前发现豆蔻酰化富含丙氨酸的C激酶底物（MARCKS），一种关键的蛋白激酶C（PKC）底物，在犬IH模型中上调。在这里，我们进一步表征的作用，MARCKS在IH和检查的表型后果MARCKS沉默的小干扰RNA（siRNA）转染在体外人血管平滑肌细胞（VSMCs）和内皮细胞（ECs），并使用快速10分钟的非病毒siRNA转染技术，以确定MARCKS沉默在体外培养的人隐静脉的影响。我们证明MARCKS沉默减弱VSMC迁移和逮捕VSMC增殖部分通过上调细胞周期蛋白依赖性激酶抑制剂p27（kip 1）。相反，MARCKS沉默对EC迁移或增殖几乎没有影响。这些表型变化最终导致培养的人隐静脉中新生内膜形成减少。这些数据确定MARCKS为IH的致病因素，并表明治疗性MARCKS沉默可以选择性抑制VSMC的“致动脉粥样硬化”增殖表型，而不会对内皮造成附带损害。这种方法可以很容易地转化为临床沉默MARCKS在静脉旁路移植术前植入。Monahan，T.美国，安徒生，北D、马丁，M。C.的方法，马利克，J.Y.，Shrikhande，G.五、普拉丹湖，Ferran，C.，LoGerfo，F. W. MARCKS沉默对人血管平滑肌和内皮细胞表型的不同影响以抑制隐静脉中的新生内膜增生。FASEB J. 23，557-564（2009）
Intimal hyperplasia (IH) limits the patency of all cardiovascular vein bypass grafts. We previously found the myristoylated alanine-rich C kinase substrate (MARCKS), a key protein kinase C (PKC) substrate, to be up-regulated in canine models of IH. Here, we further characterize the role of MARCKS in IH and examine the phenotypic consequences of MARCKS silencing by small interfering RNA (siRNA) transfection in human vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) in vitro and use a rapid 10-min nonviral siRNA transfection technique to determine the effects of MARCKS silencing in human saphenous vein cultured ex vivo. We demonstrate MARCKS silencing attenuates VSMC migration and arrests VSMC proliferation in part through the up-regulation of the cyclin-dependent kinase inhibitor p27(kip1). Conversely, MARCKS silencing had little or no effect on EC migration or proliferation. These phenotypic changes culminated in reduced neointimal formation in cultured human saphenous vein. These data identify MARCKS as a pathogenic contributor to IH and indicate therapeutic MARCKS silencing could selectively suppress the "atherogenic," proliferative phenotype of VSMCs without collateral harm to the endothelium. This approach could be readily translated to the clinic to silence MARCKS in vein bypass grafts prior to implantation.-Monahan, T. S., Andersen, N. D., Martin, M. C., Malek, J. Y., Shrikhande, G. V., Pradhan, L., Ferran, C., LoGerfo, F. W. MARCKS silencing differentially affects human vascular smooth muscle and endothelial cell phenotypes to inhibit neointimal hyperplasia in saphenous vein. FASEB J. 23, 557-564 (2009)