NCK Associated Protein 1 Modulated by miRNA-214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia.
NCK Associated Protein 1 Modulated by miRNA-214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia.
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miRNA-214 调节的 NCK 相关蛋白 1 决定血管平滑肌细胞迁移、增殖和新内膜增生
DOI：
10.1161/jaha.116.004629
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发表时间：
2016-12-07
影响因子：
5.4
通讯作者：
Xiao Q
中科院分区：
文献类型：
--
作者：
Afzal TA;Luong LA;Chen D;Zhang C;Yang F;Chen Q;An W;Wilkes E;Yashiro K;Cutillas PR;Zhang L;Xiao Q
MicroRNA miR-214参与了多种生物学功能，但miR-214及其靶基因对血管平滑肌细胞(VSMC)增殖、迁移和新生内膜平滑肌细胞增殖的影响尚不清楚。MiR-214在血管内皮细胞中的表达受不同的致病刺激通过转录机制密切调控，并且在血管损伤后表达降低。在血清缺乏的VSMC中过表达miR-214显著抑制VSMC的增殖和迁移，而敲除miR-214则显著促进VSMC的增殖和迁移。基因和蛋白质生化分析，包括蛋白质组学分析，表明NCK相关蛋白1(NCKAP1)--调节片状脂膜形成和细胞运动的波复合体的主要成分--在VSMC中受到miR-214的负调控。荧光素酶分析表明，miR-214显著抑制了VSMC中野生型荧光素酶活性，但不能抑制突变的NCKAP1 3‘非翻译区荧光素酶活性。这一结果证实了NCKAP1是miR-214在VSMC中的功能靶点。NCKAP1在VSMCs中的敲除概括了miR-214过表达对肌动蛋白聚合、细胞迁移和增殖的抑制作用。来自共转染实验的数据还显示，抑制NCKAP1对于miR-214介导的片状脂体的形成、细胞运动和生长是必需的。重要的是，局部增强的miR-214在损伤血管中的表达显著降低了NCKAP1的表达水平，抑制了VSMC的增殖，并防止了损伤后新生内膜平滑肌细胞的增殖。我们揭示了miR-214及其靶基因NCKAP1在调节VSMC功能和新生内膜增生中的重要作用。我们的发现表明miR-214是一种潜在的血管疾病治疗靶点。
MicroRNA miR‐214 has been implicated in many biological cellular functions, but the impact of miR‐214 and its target genes on vascular smooth muscle cell (VSMC) proliferation, migration, and neointima smooth muscle cell hyperplasia is unknown. Expression of miR‐214 was closely regulated by different pathogenic stimuli in VSMCs through a transcriptional mechanism and decreased in response to vascular injury. Overexpression of miR‐214 in serum‐starved VSMCs significantly decreased VSMC proliferation and migration, whereas knockdown of miR‐214 dramatically increased VSMC proliferation and migration. Gene and protein biochemical assays, including proteomic analyses, showed that NCK associated protein 1 (NCKAP1)—a major component of the WAVE complex that regulates lamellipodia formation and cell motility—was negatively regulated by miR‐214 in VSMCs. Luciferase assays showed that miR‐214 substantially repressed wild‐type but not the miR‐214 binding site mutated version of NCKAP1 3′ untranslated region luciferase activity in VSMCs. This result confirmed that NCKAP1 is the functional target of miR‐214 in VSMCs. NCKAP1 knockdown in VSMCs recapitulates the inhibitory effects of miR‐214 overexpression on actin polymerization, cell migration, and proliferation. Data from cotransfection experiments also revealed that inhibition of NCKAP1 is required for miR‐214–mediated lamellipodia formation, cell motility, and growth. Importantly, locally enforced expression of miR‐214 in the injured vessels significantly reduced NCKAP1 expression levels, inhibited VSMC proliferation, and prevented neointima smooth muscle cell hyperplasia after injury. We uncovered an important role of miR‐214 and its target gene NCKAP1 in modulating VSMC functions and neointima hyperplasia. Our findings suggest that miR‐214 represents a potential therapeutic target for vascular diseases.