TGF-β1基因转染修饰人退变腰椎间盘髓核细胞

摘要

Objective To test the possibility of modification of human degenerative lumbar disc cells by the exogenous growth factor gene, transforming growth factor β1 (TGF-β1) cDNA, and the expression of the encoded protein. Methods Nucleus pulposus samples were surgically obtained from 8 patients with degenerative lumbar disc disease. The cells were cultured and directly infected by two adenoviral constructs, Ad/CMV-EGFP containing the enhanced green fluorecence protein (EGFP) gene (marker gene) and Ad/CMV-TGF-β1 containing the potentially therapeutic TGF-β1 gene. Transgene expression was analyzed by fluorescence production and immunohistochemical staining (Ad/CMV-TGF-β1). Results Culture cells transducted by Ad/CMV-EGFP showed specific green fluorescence under the fluoroscope and expression sustained for at least 4 weeks. When infected by Ad/CMV-TGF-β1, approximally 30% of cultured cells were staind brown (+) with TGF-β1 staining. Conclusion This study established the strategy of delivering a potentially therapeutic gene, TGF-β1, by using an adenoviral vector to human degenerative lumbar intervertebral 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.