Low frequency ultrasound combined with microbubbles to promote the transfection of wild-type P53 gene in human prostate cancer cells mediated by liposome

Abstract

Objective To study low frequency ultrasound combined with microbubbles to promote the transfection of P53 gene in human prostate cancer cells mediated by liposome. Methods Ultrasound equipment was used with a frequency of 21 kHz and intensity was 46 mW/cm2 and the working time was controlled at 20% (i.e., 2 sontime and 8 sofftime) lasting 5 minutes. The human prostate cancer cell line PC-3 suspension was prepared, the cell concentration was adjusted to 1×105 cell/ ml, and cells were divided into 8 groups: control group, single microbubbles group, single ultrasound group, ultrasound combined with microbubbles group, single liposome group, liposome combined with microbubbles group, liposome combined with ultrasound group, liposome combined with ultrasound and microbubbles group. Each microbubbles group was added SonoVue 200 μl and the wild type P53 plasmid, plasmid∶liposome is 1∶2. At 24 hours after irradiation, Western-blot and RT-PCR were used to detect the gene transfection efficiency, CCK-8 was used to detect cell proliferation, then cell survival rate was calculated, cell apoptosis was detected by flow cytometry. Results After transfection, compared with single liposome group and control group, liposomes combined with ultrasound and microbubbles group can significantly improve expression of the human wild type p53 gene and protein (P<0.001). After transfection, the apoptosis rate of human prostate cancer PC-3 cells in liposomes combined with ultrasound and microbubbles group was significantly higher than that of the liposome group and control group (P<0.001). And after transfection, cell survival rate of liposome combined with ultrasound and microbubble group decreased significantly than those of single liposome group and control group (P<0.001). Conclusions Low frequency and low energy ultrasound combined with microbubbles can promote the transfection of human wild-type P53 gene mediated by liposome. Key words: Sonication; Microbubbles; Liposomes; Prostatic neoplasms