超声靶向微泡破坏联合脂质体介导pSilencer 3.1-SATB1基因转染前列腺癌细胞效果观察

摘要

Objective To investigate the effects of ultrasound-targeted microbubble destruction (UTMD) combined with liposome-mediated silencing specific nuclear matrix protein 1 gene (pSilencer3.1-SATB1) on transfection of human prostate cancer DU145 cells.Methods The DU145 cells were cultured in vitro and divided into 6 groups: the control group, microbubble group, ultrasound group, ultrasound+microbubble group, liposome group, and ultrasound+microbubble+liposome group.The control group did not do any treatment, the microbubble group was added with the appropriate amount of microbubbles, the ultrasound group only received ultrasonic irradiation, the ultrasound+microbubble group was added with microbubbles, followed by ultrasonic irradiation, the liposome group was added with liposome, and the ultrasound+microbubble+liposome group was added with microbubbles and liposomes, followed by ultrasonic irradiation.After 24-hour treatment, the expression of enhanced green fluorescent protein (EGFP) was observed by flow cytometry, the percentage of EGFP fluorescent cells was detected, and the transfection efficiency was evaluated.Results Fluorescence microscopy showed that the expression of green fluorescent protein in the ultrasound+microbubble+liposome group was numerous, which was significantly higher than that of the other groups.The transfection efficiencies of the control group, microbubble group, ultrasound group, ultrasound+microbubble group, liposome group, and ultrasound+microbubble+liposome group were 0.18%±0.09%, 0.25%±0.11%, 1.14%±0.13%, 2.96%±0.16%, 3.99%±0.12%, and 7.16%±0.17%, respectively, and the transfection efficiency of the ultrasound+microbubble+liposome group was higher than that of the other groups (all P<0.05).Conclusion UTMD can enhance the transfection efficiency of liposome-mediated plasmid pSilencer3.1-SATB1 to DUl45 cells, and improve the transfection efficiency, so it provides a new way for the treatment of prostate cancer.%目的 探讨超声靶向微泡破坏(UTMD)联合脂质体介导的沉默特异性核基质蛋白1基因(pSilencer3.1-SATB1)对人前列腺癌细胞转染的效果.方法 体外培养人前列腺癌雄激素非依赖型细胞株DU145,分为对照组、微泡组、超声组、超声+微泡组、脂质体组、超声+微泡+脂质体组.微泡组加入适量微泡,超声组仅给予超声辐照,超声+微泡组加入微泡后予以超声辐照,脂质体组加入脂质体,超声+微泡+脂质体组加入微泡、脂质体后予以超声辐照,对照组不做任何处理.24 h后采用流式细胞仪观察各组增强型绿色荧光蛋白(EGFP)表达,并检测EGFP荧光细胞的比例,计算基因转染率以评价转染效果.结果 荧光显微镜下,超声+微泡+脂质体组可见大量EGFP表达,明显多于其他各组.对照组、微泡组、超声组、超声+微泡组、脂质体组、超声+微泡+脂质体组基因转染率分别为0.18%±0.09%、0.25%±0.11%、1.14%±0.13%、2.96%±0.16%、3.99%±0.12%、7.16%±0.17%,超声+微泡+脂质体组的基因转染率高于其他各组(P均<0.05).结论 UTMD可增强脂质体介导质粒pSilencer3.1-SATB1对DU145细胞的转染效果,提高转染率,为前列腺癌治疗的研究提供了一个新的途径.