Particle-mediated gene transfer of opioid growth factor receptor cDNA regulates cell proliferation of the corneal epithelium
Particle-mediated gene transfer of opioid growth factor receptor cDNA regulates cell proliferation of the corneal epithelium
复制标题
DOI：
10.1097/01.ico.0000153561.89902.57
复制
发表时间：
2005-07-01
期刊：
影响因子：
2.8
通讯作者：
McLaughlin, PJ
中科院分区：
文献类型：
--
作者：
Zagon, IS;Sassani, JW;McLaughlin, PJ
目的：本研究旨在从分子水平上确定阿片样生长因子（OGF）和OGF受体（OGFr）之间的相互作用是否在调节角膜上皮内稳态中的DNA合成中发挥作用。将含有C-末端血凝素（HA）标记的大鼠OGFr cDNA表位的质粒pcDNA3.1+OGFr-HA或空载体pcDNA3.1+OGFr-HA与大鼠成纤维细胞生长因子受体（OGFr）进行比较。（pcDNA3.1+），通过Helios基因枪系统以300 psi向麻醉大鼠的角膜递送两次。对侧（未处理）角膜用作初始样本。结果：转染后18 h内，HA染色在角膜中央和周边的基底层和基底上层均呈阳性（阳性率> 90%）。用罗丹明偶联的抗OGFr抗体进行的定量免疫组化显示，相对于幼稚角膜，转染角膜的中央和外周上皮中的OGFr表达是幼稚角膜的两倍。BrdU阳性基底细胞的转染角膜的外周上皮细胞的数量是在幼稚corne.Conclusions的三分之一：这些数据表明，OGFOGFr系统在确定哺乳动物角膜上皮细胞更新的直接作用。此外，成功建立了一种新的递送系统的cDNA的眼表面表明，在眼睛中的基因治疗的治疗作用。
Purpose: This study was designed to determine at the molecular level whether interactions between the opioid growth factor (OGF) and OGF receptor (OGFr) play a role in regulating DNA synthesis in the homeostasis of the corneal epithelium.Methods: The plasmid pcDNA3.1+OGFr-HA, carrying the rat OGFr cDNA epitope-tagged with a C-terminal hemagglutinin (HA), or the empty-vector (pcDNA3.1+), was delivered twice by the Helios Gene Gun System at 300 psi to the cornea of anesthetized rats. The contralateral (untreated) cornea served as the naive specimen. BrdU was used to determine whether the recombinant OGFr was effective in regulating DNA synthesis in the rat peripheral corneal epithelium.Results: Within 18 hours of transfection, positive HA staining was apparent in both the basal and suprabasal layers (efficiency > 90% of the cells) throughout the central and peripheral cornea. Quantitative immunohistochemistry with rhodamine-conjugated anti-OGFr antibodies revealed twofold more OGFr expression in the central and peripheral epithelium of transfected corneas relative to naive corneas. The number of BrdU-positive basal cells in the peripheral epithelium of the transfected cornea was one-third of that in the naive cornea.Conclusions: These data demonstrate the direct role of the OGFOGFr system in determining cellular renewal in the mammalian corneal epithelium. Moreover, the successful establishment of a novel delivery system of cDNAs to the ocular surface suggests a therapeutic role for gene therapy in the eye.