超声联合声诺维辐照增强GFP基因在鼠胶质瘤C6细胞中的转染

目的 探讨超声联合声诺维微泡辐照对增强型绿色荧光蛋白基因(EGFP)在鼠胶质瘤C6细胞中的转染增效作用及安全性.方法 实验分为4组:单纯质粒组、质粒+微泡组、质粒+超声组和质粒+超声+微泡组.辐照条件为超声频率1 MHz,声强1 W/cm2,辐照时间30 s,占空比20%.按不同实验条件辐照后,应用激光共聚焦显微镜和流式细胞仪评估基因转染效率,台盼蓝染色法评估细胞活力.结果 经超声联合微泡辐照C6细胞后,GFP基因转染效率较其他组显著增加,差异有统计学意义(P<0.01),且各组均未发现显著的细胞损伤.结论 超声联合声诺维微泡辐照可显著增强GFP基因在鼠胶质瘤C6细胞的转染,为临床胶质瘤基因治疗提供了一种安全,有效的非病毒基因转染方法.

Transfection of GFP gene in rat C6 glioma cells enhanced by ultrasound-mediated microbubble destruction

Objective To determine whether ultrasound (US) exposure combined with microbubble destruction could be used to enhance non-viral gene delivery in rat C6 glioma cells. Methods Microbubbles were prepared and gently mixed with plasmid DNA. The mixture of the DNA and microbubbles was administered to cultured C6 cells under different US/microbubble conditions. US parameters adopted in this study were frequency 1 MHz, output intensity 1 W/cm2, duty cycle 20%, exposure time 30 seconds. Transfection efficiency and cell viability were assessed by FACS analysis, confocal laser scanning microscopy, and Try pan blue staining. Results Microbubble with US exposure could significantly enhance the reporter gene expression as compared with other groups. No statistical significant difference was observed in the glioma cell viability between different groups. Conclusions US-mediated microbubble destruction has the potential to promote safe and efficient gene transfer into C6 cells,and it may be useful for safe clinical gene therapy of brain cancer without a viral vector system.