A GENE-THERAPY STRATEGY USING A TRANSCRIPTION FACTOR DECOY OF THE E2F BINDING-SITE INHIBITS SMOOTH-MUSCLE PROLIFERATION IN-VIVO
A GENE-THERAPY STRATEGY USING A TRANSCRIPTION FACTOR DECOY OF THE E2F BINDING-SITE INHIBITS SMOOTH-MUSCLE PROLIFERATION IN-VIVO
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DOI：
10.1073/pnas.92.13.5855
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发表时间：
1995-06-20
影响因子：
11.1
通讯作者：
DZAU, VJ
中科院分区：
文献类型：
--
作者：
MORISHITA, R;GIBBONS, GH;DZAU, VJ
应用DNA技术在体内调控疾病相关基因的转录具有重要的治疗潜力，转录因子E2 F在参与血管损伤后病变形成的细胞周期调控基因如c-myc、cdc 2和编码增殖细胞核抗原（PCNA）的基因的协同反式激活中起关键作用，我们假设，与E2 F的高亲和力的双链DNA可以引入体内作为诱饵结合E2 F，并阻断介导细胞周期进展和血管损伤后内膜增生的基因的激活。凝胶迁移率变化分析显示E2 F诱饵对E2 F结合蛋白的完全竞争。E2 F decoy转染可抑制c-myc、cdc 2和PCNA基因的表达以及血管平滑肌细胞的增殖。体内转染后2周，E2 F decoy可显著抑制新生内膜的形成，并以剂量依赖性方式持续至单次转染后8周。因此，E2 F诱饵的转移可以在体内调节基因表达并抑制平滑肌增殖和血管病变形成。
The application of DNA technology to regulate the transcription of disease-related genes in vivo has important therapeutic potentials, The transcription factor E2F plays a pivotal role in the coordinated transactivation of cell cycle-regulatory genes such as c-myc, cdc2, and the gene encoding proliferating-cell nuclear antigen (PCNA) that are involved in lesion formation after vascular injury, We hypothesized that double-stranded DNA with high affinity for E2F may be introduced in vivo as a decoy to bind E2F and block the activation of genes mediating cell cycle progression and intimal hyperplasia after vascular injury, Gel mobility-shift assays showed complete competition for E2F binding protein by the E2F decoy. Transfection with E2F decoy inhibited expression of c-myc, cdc2, and the PCNA gene as well as vascular smooth muscle cell proliferation both in vitro and in the in vivo model of rat carotid injury, Furthermore, 2 weeks after in vivo transfection, neointimal formation was significantly prevented by the E2F decoy, and this inhibition continued up to 8 weeks after a single transfection in a dose-dependent manner. Transfer of an E2F decoy can therefore modulate gene expression and inhibit smooth muscle proliferation and vascular lesion formation in vivo.