Department of Basic Oncology, Institute of Oncology, Hacettepe University, 06100, Sihhiye, Ankara, Turkey,
Cytotechnology. 2009 Nov;61(1-2):45-53. doi: 10.1007/s10616-009-9241-9. Epub 2009 Dec 6.
Gene transfer into haematopoietic cells is a challenging approach. The extracellular matrix component fibronectin has been known to modulate the cell cycle dynamics, viability and differentiation of leukaemia cells. Thus, our aim was to investigate the influence of fibronectin substratum on the liposomal transfection of myeloid leukaemia cell lines. Liposomal transfection was performed with K562 and HL-60 as representative lines of transfection-competent and -incompetent myeloid leukaemia cells, respectively. Flow cytometry analyses were performed to determine transfection efficiency monitored by green fluorescent protein (GFP) expression and to assess cell viability and cell cycle status. Quantitation of GFP gene expression and DNA uptake was assayed by real time PCR. The current data showed that the adhesion to fibronectin deteriorated the transfection of K562 cells. In contrary, it enhanced the delivery of plasmid DNA into HL-60 cells. Correspondingly, the adhesion to fibronectin influenced the transfection efficiency mainly by modulating the intracellular presence of plasmid DNA. The cell cycle and viability which is regulated by fibronectin had a minor impact on the success of gene delivery. This phenomenon may be considered as an important factor which may modulate the potential gene transfer approaches for myeloid leukaemia.
基因转移到造血细胞是一种具有挑战性的方法。细胞外基质成分纤维连接蛋白已被证明可以调节白血病细胞的细胞周期动态、活力和分化。因此,我们的目的是研究纤维连接蛋白基质对髓系白血病细胞系脂质体转染的影响。脂质体转染采用 K562 和 HL-60 作为转染能力和转染能力差的髓系白血病细胞的代表性系进行。通过流式细胞术分析确定通过绿色荧光蛋白 (GFP) 表达监测的转染效率,并评估细胞活力和细胞周期状态。通过实时 PCR 定量测定 GFP 基因表达和 DNA 摄取。目前的数据表明,纤维连接蛋白的黏附降低了 K562 细胞的转染。相反,它增强了质粒 DNA 向 HL-60 细胞的传递。相应地,纤维连接蛋白的黏附主要通过调节细胞内质粒 DNA 的存在来影响转染效率。由纤维连接蛋白调节的细胞周期和活力对基因传递的成功影响较小。这种现象可能被认为是调节髓系白血病潜在基因转移方法的一个重要因素。