A potential approach for gene therapy targeting hepatoma using a liver-specific promoter on a retroviral vector.-猫眼课题宝

使用逆转录病毒载体上的肝脏特异性启动子进行针对肝癌的基因治疗的潜在方法。

DOI：

10.1247/csf.16.503

发表时间：

1991

期刊：

Cell structure and function

影响因子：

1.5

通讯作者：

中科院分区：

生物学4区

文献类型：

作者：

S. Kuriyama;M. Yoshikawa;S. Ishizaka;T. Tsujii;K. Ikenaka;T. Kagawa;N. Morita;K. Mikoshiba

最近在分子生物学和人类及其他哺乳动物细胞的体外培养方面取得的技术进步，使医学和科学界广泛接受基因治疗遗传疾病的可行性。癌症实际上可能是这种治疗的有吸引力的目标之一。对于癌症的治疗，重要的是操纵感兴趣的基因，使其仅在癌细胞中表达。我们已经开发了一个组织特异性的基因表达系统，基于逆转录病毒载体上的组织特异性启动子。构建了小鼠亲嗜性逆转录病毒载体，其中大肠杆菌β-半乳糖苷酶基因作为报告基因;它在白蛋白增强子元件和启动子的控制下表达。首先在体外评估该载体的组织特异性，并且仅在肝癌细胞系中检测到β-半乳糖苷酶活性。将该重组逆转录病毒直接注射到由可移植的小鼠MH-134肝癌细胞组成的皮下肿瘤中，并在体内观察该基因的表达。然后将该重组逆转录病毒经脾或直接注射入肝，导致部分肝切除小鼠的分裂肝细胞中的基因表达，但在正常小鼠的非分裂肝细胞中不表达。通过逆转录病毒载体转移特异于分裂肝细胞的基因和表达，应该具有选择性消除被非分裂正常肝细胞包围的肝癌细胞的高潜力。

Recent technological advances made in molecular biology and in vitro culture of human and other mammalian cells have led to broad medical and scientific acceptance of the feasibility of gene therapy for genetic diseases. Cancer might practically be one of the attractive targets for such therapy. For the treatment of cancer, it is important to manipulate the gene of interest such that it is expressed solely in cancer cells. We have developed a tissue-specific gene expression system, based on a tissue-specific promoter on a retroviral vector. A murine ecotropic retroviral vector was constructed in which the Escherichia coli beta-galactosidase gene served as a reporter; it was expressed under control of the albumin enhancer element and promoter. The tissue specificity of this vector was first assessed in vitro, and beta-galactosidase activity was detected exclusively in hepatoma cell lines. This recombinant retrovirus was injected directly into a subcutaneous tumor composed of transplantable murine MH-134 hepatoma cells, and expression of the gene was observed in vivo. Then this recombinant retrovirus was injected via the spleen or directly into the liver, resulting in the gene expression in dividing hepatocytes in partially hepatectomized mice, but not in nondividing hepatocytes in normal mice. Gene transfer specific to dividing hepatocytes and expression by means of retroviral vectors should possess high potential for selective elimination of hepatoma cells surrounded by nondividing normal hepatocytes.