Nishi T, Yoshizato K, Yamashiro S, Takeshima H, Sato K, Hamada K, Kitamura I, Yoshimura T, Saya H, Kuratsu J, Ushio Y
Department of Neurosurgery, Kumamoto University School of Medicine, Honjo, Kumamoto, Japan.
Cancer Res. 1996 Mar 1;56(5):1050-5.
A novel method for high-efficiency and region- controlled in vivo gene transfer was developed by combining in vivo electroporation and intraarterial plasmid DNA injection. A mammalian expression plasmid for the Escherichia coli lacZ gene (driven with a SV40 early promoter) was injected into the internal carotid artery of rats whose brain tumors (from prior inoculation) had been electroporated between two electrodes. The lacZ gene was efficiently transferred and expressed in the tumor cell 3 days after plasmid injection. However, neither any gene transfers nor any elevated lacZ activity was detected in tissues outside the electrodes. The plasmid was not transferred without electroporation. Human monocyte chemoattractant protein-1 cDNA was also transferred by this method, and its long-lasting (3 weeks) expression was confirmed by using the Epstein-Barr virus episomal replicon system. The expressed monocyte chemoattractant protein-1 protein was functional, as evident by the presence of a large number of monocytes in tumor tissue. This method, electrogene therapy, which does not require viral genes or particles, allows genes to be transferred and expressed in desired organs or tissues, and it may lead to the development of a new type of highly effective gene therapy.
通过将体内电穿孔与动脉内注射质粒DNA相结合，开发出一种用于高效且区域可控的体内基因转移的新方法。将一种用于大肠杆菌lacZ基因（由SV40早期启动子驱动）的哺乳动物表达质粒注入大鼠的颈内动脉，这些大鼠的脑肿瘤（先前接种所致）已在两个电极之间进行了电穿孔处理。质粒注射3天后，lacZ基因在肿瘤细胞中得到高效转移并表达。然而，在电极之外的组织中未检测到任何基因转移或lacZ活性升高。未进行电穿孔时，质粒不会转移。人单核细胞趋化蛋白-1 cDNA也通过该方法进行了转移，并使用爱泼斯坦-巴尔病毒附加体质粒复制系统证实了其持久（3周）表达。表达的单核细胞趋化蛋白-1具有功能，肿瘤组织中存在大量单核细胞即证明了这一点。这种方法，即电基因疗法，不需要病毒基因或病毒颗粒，能使基因在期望的器官或组织中转移并表达，可能会引领新型高效基因疗法的发展。
