酪氨酸羟化酶和GTP环水解酶-1基因修饰永生化成纤维细胞及其表达研究

Expression of immortalized fibroblasts genetically modified by tyrosine hydroxylase gene and GTP cyclohydrolase-1 gene

目的 探讨酪氨酸羟化酶 (TH)和GTP环水解酶 1(GCH)基因修饰的永生化成纤维细胞的可行性和表达能力 ,为其混合移植治疗帕金森病 (PD)作准备。方法 SV40大T抗原转化成纤维细胞使其永生化。构建TH和GCH基因真核表达质粒并转染永生化成纤维细胞 ,利用免疫组化和RT PCR检测其在体内外的表达。结果 TH和GCH基因修饰的永生化成纤维细胞在体外可长期传代培养且可有效表达 ,脑内移植后TH基因至少可表达 8周以上。

Objective To explore the expression of tyrosine hydroxylase(TH) and GTP cyclohydrolase 1 (GCH) gene in genetically modified immortalized fibroblasts for potential use in the treatment of Parkinson's disease (PD). Methods Rat fibroblasts were immortalized by SV40 large antigen transformation. Eukaryotic plasmid vector encoding TH gene or GCH gene was constructed and introduced into the fibroblasts respectively, and immunohistochemistry or RT PCR was performed to identify the exogeneous gene expression in vitro and in vivo . Results Immortalized fibroblasts genetically modified by TH gene and GCH gene could survive long termly and express the exogenous gene in vitro . When the gene modified fibroblasts were intracerebrally transplanted, TH gene was expressed for at least 8 weeks without tumor forming or immune repelling phenomena. Conclusion Immortalized fibrblasts genetically modified by TH and GCH gene may be developed as a useful therapy for PD.