低频超声联合微泡造影剂增强脂质体介导pEGFP-N1基因转染体外前列腺癌细胞

目的探讨低频超声联合微泡造影剂增强脂质体介导增强型绿色荧光蛋白质粒(pEGFP-N1)转染前列腺癌细胞的可行性,并对超声微泡浓度参数进行优化。方法将前列腺癌PC-3细胞悬液分为空白对照组、超声组、微泡组、微泡+超声组、脂质体组、脂质体+微泡组、脂质体+超声组、脂质体+微泡+超声组,其中脂质体+微泡+超声组根据微泡体积浓度不同分为(0、10%、20%、30%、40%和50%)6个亚组。经超声辐照,24h后用荧光显微镜观察细胞中基因表达情况,并用流式细胞仪检测转染率。结果脂质体+微泡+超声组基因转染效率最高,与其他组比较差异均有统计学意义(P均<0.05);在脂质体+微泡+超声亚组中,微泡浓度为20%亚组基因转染率最高。结论低频超声联合微泡能有效增强脂质体介导pEGFP-N1基因在体外前列腺癌细胞中的转染率,20%是体外基因转染前列腺癌细胞的最佳微泡浓度。

Liposome enhanced transfection of pEGFP-N1 plasmid to prostate cancer cells in vitro with low frequency ultrasound combined with microbubbles

Objective To investigate the feasibility of liposome enhanced transfection of green fluorescent protein gene (pEGFP-N1) plasmid to prostate cancer cells with low frequency ultrasound combined with microbubbles, and to optimize the parameters of microbubbles concentration. Methods PC-3 prostate cancer cell suspension was divided into 8 groups, i.e. control group, ultrasound group, microbubbles group, microbubbles+ultrasound group, liposome group, microbubbles+liposome group, liposome+ultrasound group and ultrasound+microbubbles+liposome group. The ultrasound+microbubbles+liposome group was classified into 6 sub-groups: 0, 10%, 20%, 30%, 40%, and 50%, based on microbubbles volume concentration. The cell suspension was cultured in 12-well plates for 24 h after irradiation, and fluorescent microscopy was used to observe gene transfection and calculated the rate of gene transfection. Results Ultrasound+microbubbles+liposome group had the best efficiency, which was significantly different compared with the other groups (all P<0.05), while 20% microbubbles concentration sub-group had the highest rate of gene transfection in the ultrasound+microbubbles+liposome group. Conclusion Low-frequency ultrasound in combination with microbubbles can significantly enhance liposome-mediated in vitro pEGFP-N1 gene transfection rate. For microbubbles concentration, 20% is the best.