绿色荧光蛋白基因在白血病细胞K562中的转导效率

目的：探讨慢病毒载体在白血病细胞中的基因转导效率，为白血病基因治疗提供关键依据。方法：应用1型人类免疫缺陷病毒(HIV-1)改造而成的第3代自身失活(SIN）慢病毒载体（lentiviral vector）系统，与鼠白血病病毒（MLV）SIN载体进行比较，通过荧光显微镜观察细胞内标记基因绿色荧光蛋白（GFP）的表达情况，应用流式细胞术检测基因导入细胞百分比，评价两种载体系统在人白血病细胞系K562中的基因转导效率。结果：通过荧光显微镜可定性观察GFP在K562细胞中的表达情况，在同样的基因转导条件下，HIV载体转导的白血病细胞中被转导的标记基因GFP的表达强度及GFP阳性细胞数明显高于MLV载体转导的细胞。流式细胞仪定量检测HIV载体的转导效率接近100%，而MLV低于40%,两组间转导效率比较差异有显著性（P＜0.05）。结论：基于慢病毒载体基因转导的高效性，该载体系统可作为白血病细胞基因转导的极好工具。

Transduction efficiency of green fluorescent protein gene in leukemia cells K562

Objective To investigate the gene transduction efficiency of lentiviral vector in leukemia cells to provide key basis for leukemia gene therapy. Methods A third-generation self-inactivating(SIN) lentiviral vector system based on human immunodeficiency virus type 1(HIV-1) was used to improve transduction efficiency.The transduction efficiency of the HIV-1-based vector was compared directly with the moloney murine leukemia virus(MLV) SIN vector in human leukemia cell line K562.The expression of green fluorescent protein(GFP) in cells was observed by fluorescence microscopy and flow cytometry(FCM) to detect the percentage of gene trasduction cells.Results The GFP expression in K562 cells was observed qualitatively by fluorescence microscopy.At the same gene transduction conditions,the GFP marker gene expression intensity and GFP positive cells in leukemia cells transduced with HIV vectors were significantly higher than those transduced with MLV vectors.Initial transduction efficiencies were almost 100% for the HIV and less than 40% for the MLV vectors. The transduction efficiency had significant difference between HIV vector group and MLV group(P<0.05).Conclusion This lentiviral vector is an excellent gene transduction system for leukemia cells because of its high gene transduction efficiency.