Forkhead transcription factor FOXO3a is a negative regulator of angiogenic immediate early gene CYR61, leading to inhibition of vascular smooth muscle cell proliferation and neointimal hyperplasia
Forkhead transcription factor FOXO3a is a negative regulator of angiogenic immediate early gene CYR61, leading to inhibition of vascular smooth muscle cell proliferation and neointimal hyperplasia
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DOI：
10.1161/01.res.0000257945.97958.77
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发表时间：
2007-02-16
影响因子：
20.1
通讯作者：
Kim, Hyo-Soo
中科院分区：
文献类型：
--
作者：
Lee, Hae-Young;Chung, Jae-Woong;Kim, Hyo-Soo
富含半胱氨酸的血管生成蛋白 61（CYR61，CCN1）是一种在生长因子刺激下在血管平滑肌细胞（VSMC）中表达的立即早期基因，其表达被认为与血管成形术后再狭窄有关。据报道，叉头转录因子在细胞增殖、细胞凋亡、甚至适应细胞应激中发挥多种作用。我们假设叉头转录因子FOXO3a可能调节VSMC中CYR61的表达，并研究FOXO3a在血管对血管活性信号的反应和血管损伤过程中的CYR61调节作用。为了评估 FOXO3a 对 CYR61 表达的影响，用表达组成型活性 FOXO3a 的腺病毒载体 (Ad-TM-FOXO3a) 感染大鼠 VSMC。持续活跃的 FOXO3a 基因转导抑制了 CYR61 的表达。使用 CYR61 启动子区域叉头因子结合基序的缺失构建体进行荧光素酶测定，与完整构建体相比，导致荧光素酶表达显着降低，染色质免疫沉淀分析证实了 FOXO3a 对 CYR61 的转录调节。血清和血管紧张素 II 快速诱导 CYR61 表达，Ad-TM-FOXO3a 显着降低 CYR61 表达。共转染表达 CYR61 的腺病毒载体可显着逆转与 FOXO3a 激活相关的 VSMC 增殖和迁移减少，而 FOXO3a 诱导的细胞凋亡不受影响。在大鼠颈动脉球囊损伤模型中，CYR61在损伤早期在VSMC中快速诱导，并保持升高状态直至14天，而Ad-TM-FOXO3a转染可抑制CYR61的升高。 14 天后，与对照组相比，FOXO3a 转导后的新内膜显着减少（0.06 +/- 0.02 对比 0.20 +/- 0.07 mm(2)，P < 0.01）。 Ad-TM-FOXO3a 对内膜增生的这种减少可通过补充 CYR61 来逆转。这些数据表明 FOXO3a 是 CYR61 的负转录因子，并且抑制 CYR61 是 FOXO3a 抑制 VSMC 增殖和新内膜增生的多种机制之一。
Cysteine-rich angiogenic protein 61 (CYR61, CCN1) is an immediate early gene expressed in vascular smooth muscle cells (VSMCs) on growth factor stimulation, and its expression has been suggested to be associated with postangioplasty restenosis. The forkhead transcription factors are reported to play various roles in cellular proliferation, apoptosis, and even adaptation to cellular stress. We hypothesized that the forkhead transcription factor FOXO3a may regulate CYR61 expression in VSMCs and investigated the CYR61-modulating effect of FOXO3a in the process of vascular response to vasoactive signals and vascular injury. To evaluate the effect of FOXO3a on CYR61 expression, rat VSMCs were infected with adenoviral vectors expressing constitutively active FOXO3a (Ad-TM-FOXO3a). Constitutively active FOXO3a gene transduction suppressed CYR61 expression. Luciferase assay with the deletion constructs of the forkhead factor binding motif in CYR61 promoter region, which resulted in a significant decrease in luciferase expression compared with the intact construct, and chromatin immunoprecipitation analysis confirmed transcriptional regulation of CYR61 by FOXO3a. Serum and angiotensin II rapidly induced CYR61 expression, which was significantly reduced by Ad-TM-FOXO3a. Reduction of VSMC proliferation and migration associated with FOXO3a activation was significantly reversed by cotransfection of adenoviral vector expressing CYR61, whereas apoptosis induction by FOXO3a was not influenced. In a rat balloon carotid arterial injury model, CYR61 was rapidly induced in VSMCs in the early stage of injury and remained elevated until 14 days, which was suppressed by Ad-TM-FOXO3a transfection. After 14 days, there was a significant reduction in neointima by FOXO3a transduction compared with the control group (0.06 +/- 0.02 versus 0.20 +/- 0.07 mm(2), P < 0.01). Such reduction of neointimal hyperplasia by Ad-TM-FOXO3a was reversed by CYR61 replenishment. These data suggest that FOXO3a is a negative transcription factor of CYR61 and that suppression of CYR61 is among several mechanisms by which FOXO3a inhibits VSMC proliferation and neointimal hyperplasia.