超声辐照SonoVue增强脂质体介导pEGFP-N1转染乳腺癌细胞

目的 探讨超声辐照SonoVue介导增强型绿色荧光蛋白报告基因质粒(pEGFP-N1)转染乳腺癌MCF-7细胞的最优条件以及超声联合微泡造影剂增强脂质体转染的效果.方法 体外培养人乳腺癌MCF-7细胞,按照不同的辐照条件分组:不处理组,超声辐照十质粒组,微泡浓效组,超声声张组,辐照时间组及工作周期组.分别进行超声辐照,辐照后以MTT法测细胞存活度,流式细胞仪测pEGFP-N1瞬时转染率,取得最佳的辐照条件.重新准备细胞后先以脂质体转染细胞,2 h后加入超声微泡并以最佳条件辐照,用同样的方法测细胞存活率及转染率.结果 20%的造影剂浓度、1.5 W/cm2的声强、90 s的辐照时间、20%的工作周期(占空比)为超声辐照SonoVue介导pEGFP-N1转染MCF-7细胞的最优条件,转染率为(14.21±2.55)%,细胞存活率为(84.60±2.85)%.在增强脂质体转染的实验中,超声微泡脂质体组的转染率最高(22.15±2.69)%,与其他各组相比差异有统计学意义(P<0.05),此时细胞存活率为(80.13±3.61)%.结论 20%的造影剂浓度、1.5 W/cm2的声强、90 s的辐照时间、20%的工作周期是MCF-7细胞的最佳转染条件,超声联合微泡能够增强脂质体转染的效果.

Liposome enhanced transfection of pEGFP-N1 plasmid to MCF-7 in vitro with SonoVue combined with ultrasound

Objective To investigate the optimal irradiation condition of SonoVue combined with ultrasound mediated gene transfection to MCF-7 in vitro, and to observe the efficiency of liposome transfection enhanced by SonoVue combined with ultrasound. Methods The MCF-7 were cultured in 24-well dishes and all cells were divided into 6 groups: no-treatment group, plasmid and ultrasound group, contrast concentration group, ultrasound intensities group, ultrasound radiation time group, and duty cycle group. After radiation, gene transfer was quantified with flow cytometry, and the survival rate of cells was calculated with MTT. Then the cells were cultured again and divide into 4 groups: liposome group, ultrasound and SonoVue group, ultrasound and liposome group, ultrasound combined with SonoVue and liposome group. The transfection rate and the survival rate of cells were calculated with the method mentioned above. Results In the first 6 groups, the optimal irradiation condition of SonoVue with ultrasound mediated gene transfection was 20% SonoVue, 1.5 W/cm², 20% duty cycle and 90 s exposure, transfection rate was 14.21%±2.55%, and survival rate was 84.60%±2.85%. In the study of enhancing efficiency of liposome transfection, the group of ultrasound combined with SonoVue and liposome had the highest transfection rate of 22.15%±2.69%, statistically different from other groups (P<0.05), and the survival rate was 80.13%±3.61%. Conclusion The optimal irradiation condition are 20% SonoVue, 1.5 W/cm², 20% duty cycle and 90 s exposure. SonoVue combined with ultrasound can enhance the efficiency of liposome transfection.