Morishita R, Gibbons G H, Horiuchi M, Ellison K E, Nakama M, Zhang L, Kaneda Y, Ogihara T, Dzau V J
Division of Cardiovascular Medicine, Falk Cardiovascular Research Center, Stanford University School of Medicine, CA 94305-5246, USA.
Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5855-9. doi: 10.1073/pnas.92.13.5855.
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.
将DNA技术应用于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E2F在细胞周期调控基因（如c-myc、cdc2以及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，这些基因参与血管损伤后病变的形成。我们推测，对E2F具有高亲和力的双链DNA可作为诱饵引入体内，以结合E2F并阻断血管损伤后介导细胞周期进程和内膜增生的基因的激活。凝胶迁移率变动分析显示，E2F诱饵与E2F结合蛋白完全竞争。在大鼠颈动脉损伤的体外和体内模型中，用E2F诱饵转染可抑制c-myc、cdc2和PCNA基因的表达以及血管平滑肌细胞的增殖。此外，在体内转染2周后，E2F诱饵可显著预防新生内膜形成，且这种抑制在单次转染后持续长达8周，呈剂量依赖性。因此，E2F诱饵的转导可在体内调节基因表达并抑制平滑肌增殖和血管病变形成。
