Schakowski Frank, Buttgereit Peter, Mazur Martin, Märten Angela, Schöttker Björn, Gorschlüter Marcus, Schmidt-Wolf Ingo GH
Medizinische Klinik und Poliklinik I, Rheinische Friedrich-Wilhelms-Universität, Bonn, Germany.
Genet Vaccines Ther. 2004 Jan 12;2(1):1. doi: 10.1186/1479-0556-2-1.
Tumor cells such as leukemia and lymphoma cells are possible targets for gene therapy. However, previously leukemia and lymphoma cells have been demonstrated to be resistant to most of non-viral gene transfer methods. METHODS: The aim of this study was to analyze various methods for transfection of primary leukemia cells and leukemia cell lines and to improve the efficiency of gene delivery. Here, we evaluated a novel electroporation based technique called nucleofection. This novel technique uses a combination of special electrical parameters and specific solutions to deliver the DNA directly to the cell nucleus under mild conditions. RESULTS: Using this technique for gene transfer up to 75% of primary cells derived from three acute myeloid leukemia (AML) patients and K562 cells were transfected with the green flourescent protein (GFP) reporter gene with low cytotoxicity. In addition, 49(+/- 9.7%) of HL60 leukemia cells showed expression of GFP. CONCLUSION: The non-viral transfection method described here may have an impact on the use of primary leukemia cells and leukemia cell lines in cancer gene therapy.
白血病和淋巴瘤细胞等肿瘤细胞是基因治疗的潜在靶点。然而，此前已证明白血病和淋巴瘤细胞对大多数非病毒基因转移方法具有抗性。
本研究的目的是分析原代白血病细胞和白血病细胞系的多种转染方法，并提高基因传递效率。在此，我们评估了一种基于电穿孔的新技术——核转染。这种新技术结合了特殊的电参数和特定溶液，在温和条件下将DNA直接递送至细胞核。
使用该技术进行基因转移，来自三名急性髓系白血病（AML）患者的原代细胞和K562细胞中高达75%用绿色荧光蛋白（GFP）报告基因进行了转染，且细胞毒性较低。此外，49（±9.7%）的HL60白血病细胞显示出GFP表达。
本文所述的非病毒转染方法可能会对原发性白血病细胞和白血病细胞系在癌症基因治疗中的应用产生影响。
