摘要
根据细胞因子之间协同作用的特点，采用重组 Ｄ Ｎ Ａ 技术设计并构建了人 Ｉ Ｌ２ 与 Ｉ Ｆ Ｎα融合基因，并用肝癌组织特异的 Ａ Ｆ Ｐ增强子／ Ａ Ｌ Ｂ启动子调控融合基因在肝癌细胞中的靶向表达．实验结果表明，克隆的 Ｅ Ａ Ｆ Ｐ Ｐ Ａ Ｌ Ｂ联合转录调控序列能调控细胞因子基因在 Ａ Ｆ Ｐ阳性人肝癌细胞中靶向表达， Ｉ Ｌ２／ Ｉ Ｆ Ｎα２ｂ 融合基因的表达水平与感染肝癌细胞的 Ａ Ｆ Ｐ表达水平呈正相关性．实验证明表达的融合蛋白具有 Ｉ Ｌ２ 和 Ｉ Ｆ Ｎ 两种生物学活性的细胞因子．这可能为肝癌基因治疗开辟新途径．
The study explored the use of a tumor specific AFP enhancer and a liver specific albumin promoter to achieve regulated cytokine IL 2/IFNα2b fused gene expression for treatment of hepatocellular carcinoma (HCC).The human AFP enhancer(E AFP )/albumin promoter(P ALB )were amplified from human chromosme DNA by PCR method.A recombinant retrovirus including the selectable marker neo R gene and IL 2/IFNα2b fused gene controled by the E AFP P ALB was constructed.The liver targeted expression pattern of the IL 2/IFNα2b fused gene was observed when the product was tested in culture medium of the infected cells.IL 2 activity was 850 IU/10 6 cells,IFNα activity was 320 IU/10 6 cells.The results showed that E AFP P ALB combind transcriptional regulatory sequences could control the targeted expression of cytokine genes in AFP positive human hepatoma cells,and the expression level of IL 2/IFNα2b fused gene was positively correlated to the level of AFP expression in the infected cells.The expressed IL 2/IFNα2b fused protein had functions of both IL 2 and IFNα.Therefore,this study illustrates the superiority of using transcriptionally targeted recombinant retrovirus vectors in cytokine based gene therapy.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第4期636-641,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金
关键词
IFNΑ
融合基因
AFP增强子
肝癌
血细胞介素2
Human IL 2/IFNα2b fused gene,Human AFP enhancer,Human albumin promoter,Targeted expression,Hepatocellular carcinoma
