脉冲超声辐照微泡增强脂质体介导基因转染的体外实验研究

目的 探讨脉冲超声辐照微泡超声造影剂提高脂质体介导增强型绿色荧光蛋白质粒在肝癌,细胞(HepG2)中转染率的可行性.方法 将HepG2接种在24孔板中,随机分为以下6组:①空白对照组;②质粒+脂质体组;③超声+质粒组;④超声+质粒+微泡组;⑤超声+质粒+脂质体组;⑥超声+质粒+脂质体+微泡组.荧光显微镜及流式细胞仪检测增强型绿色荧光蛋白表达及转染效率;MTT法检测此转染方法对HepG2生长活性的影响.结果 超声+质粒+脂质体组的转染效率最高,并对细胞活性无明显影响;而超声+质粒+脂质体+微泡组转染效率与超声+质粒+脂质体组比较差异无统计学意义(P＞0.05),但细胞活性与空白组比较差异有统计学意义(P＜0.05).结论 脉冲超声辐照可介导基因转染,并能提高脂质体基因转染效率.同时,在一定条件下,超声微泡造影剂可提高超声介导基因转染效率,此方法可为基因转染提供新的思路.

The study of pulsed-ultrasound destructed microbubble to enhance liposome gene transfection in vitro

Objective To investigate whether pulsed ultrasound-mediated microbubbles destruction could effectively enhance the efficiency of liposome delivery plasmid to hepatoma cells.Methods The cultured HepG2 cells were divided into six groups.The first group was as contrast group and the second group was given proper dose of liposome and plasmid.Pulsed-ultrasound exposed liposome was the third group.Add plasmid and microbubbles to the fourth group and exposed to pulsed-ultrasound.Add plasmid and liposome to the fifth group and exposed to pulsed-ultrasound,too.Liposome with plasmid and microbubbles were applied to the sixth group and exposed to ultrasound.After 24 hours,the EGFP expression in the hepatoma cells was detected by fluorescence microscopy and MTT.Results Green fluorescence intensity and transfection efficiency of the fifth group were higher than that of other groups,and cell viability had no significant difference.Although they were high in the sixth group,cells were partly died.Conclusions The liposome delivered EGFP expression efficiency in hepatoma cells was increased with the administration of pulsed ultrasound-mediated microbubbles destruction.In certain condition,microbubbles can enhance the efficiency of plasmid.