Adenovirus-mediated TGF-β1 gene transfer to human degenerative lumbar intervertebral disc cells  

TGF-β_1基因转染修饰人退变腰椎间盘髓核细胞

作　　者：赵剑[1] 贾连顺[1] 陈德玉[1] 肖剑[1] 潘欣[1] 荆鑫[1] 

机构地区：[1]第二军医大学附属长征医院骨科,上海200003

出　　处：《Chinese Medical Journal》2002年第3期409-412,共4页中华医学杂志（英文版）

摘　　要：Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β_1 (TGF-β_ 1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerat ive lumbar disc disease. The cells were cultured and directly infected by two a denoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence pro tein (EGFP) gene (marker gene) and Ad/CMV-TGF-β_1 containing the potentially therapeutic TGF-β_1 gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-β_1). Results Culture cells transducted by Ad/CMV-EGFP showed specific green fluorescence und er the fluoroscope and expression sustained for at least 4 weeks. When infe cted by Ad/CMV-TGF-β_1, approximally 30% of cultured cells were staind brown (+) with TGF-β_1 staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene , TGF-β_1, by using an adenoviral vector to human degenerative lumbar interve rtebral disc cells.目的　探讨对人退变腰椎间盘细胞以外源性生长因子TGF β1进行基因修饰的可能性 ,同时检测相应编码蛋白的表达情况。方法　髓核细胞取自 8例因腰椎间盘退变手术病人 ,在体外进行原代培养 ,并分作 3组。将构建好的分别携带增荧光绿色蛋白EGFP基因 (报告基因 )和TGF β1基因的腺病毒载体Ad/CMV EGFP、Ad/CMV TGF β1直接转染细胞 ,对转染情况进行观察。结果　第一组 (Ad/CMV EGFP转染组 )经荧光显微镜下观察有特异性绿色荧光表达 ,并持续 4周 ,表示外源基因转入成功 ;第二组 (Ad/CMV TGF β1转染组 )经组化染色证明产生TGF β1,转染率约 30 % ,而作为对照的第三组则染色基本阴性。结论　采用腺病毒载体可以将TGF β1基因转染修饰腰椎退变椎间盘细胞 ,并能表达相应的生长因子TGF β1,为今后腰椎间盘退变疾病的基因治疗提供了实验基础。

关 键 词：adenovirus · degenerative disc cell · lum bar intervertebral disc · growth factor · gene therapy 

分 类 号：R681.53[医药卫生—骨科学]