逆转录病毒载体携带报告基因在色素上皮细胞的转染与表达  

Transfection and expression of retrovirus-mediated reporter genes in retinal pigment epithelial cells in vitro

作　　者：万光明[1] 邢杉杉[1] 

机构地区：[1]郑州大学第一附属医院眼科,河南省郑州市450052

出　　处：《眼科新进展》2008年第2期94-95,99,共3页Recent Advances in Ophthalmology

基　　金：河南省杰出青年基金资助(编号:512000600)~~

摘　　要：目的观察逆转录病毒载体携带报告基因在视网膜色素上皮（retinal pigment epithelium，RPE）细胞的表达情况，以探讨基因疗法用于抑制增生性玻璃体视网膜病变的可行性。方法选第3～4代培养的已长满融合的RPE细胞，以1：3的分种率分种传代。随机分为对照组与实验组，每组12孔。实验组用绿色荧光蛋白基因作为报告基因，以逆转录病毒载体携带来转染RPE细胞，重组合绿色荧光蛋白基因的逆转录病毒上清液浓度为1．2×10^9efu·L^-1对照组RPE细胞以同样方法直接转染绿色荧光蛋白基因质粒DNA。观察绿色荧光蛋白基因在2组RPE细胞的表达情况。结果基因转染后可见实验组RPE细胞的绿色荧光蛋白基因表达率可达56％-72％，对照组未见有任何表达。结论逆转录病毒载体可以携带目的基因转染于RPE细胞，并达到较高的转染率。[眼科新进展2008；28（2）：94-95，99]Objective To observe the expression of retrovirus-mediated re- porter genes in retinal pigment epithelial（RPE） cells and discuss the possibility of gene therapy to inhibit the proliferative vitreoretinopathy. Methods RPE ceUs,confluenced at the 3rd or 4th generation,were passaged at 1 ： 3 and divided into control group and experimental group,12 holes in each group. In experimental group,green fluorescence protein （GFP） genes served as reporter genes and were taken by retrovirus genes. Supernatant liquid of retrovirus with GFP was 1.2 × 10^9 cfu · L^-1. In control group, RPE trabsfered GFP DNA plasmid directly in the same way, and GFP expression was observed in 2 groups. Results Rate of green fluorescence protein gene expression was from 56% to 72%. No GFP gene expression was seen in control group. Conclusion Retrovirus-mediated gene can be transfered into RPE cells efficiently. [ Rec Adv Ophthalmol 2008;28（ 2 ） ：94-95,99 ]

关 键 词：视网膜色素上皮细胞 增生性玻璃体视网膜病变 报告基因 基因治疗 

分 类 号：R774.1[医药卫生—眼科]