【摘要】 目的探讨超声靶向微泡破碎(ultrasound targeted microbubble destruction,UTMD)介导EGFP质粒转染肝癌细胞株HepG2的有效性、安全性并优化超声辐照参数。方法体外培养HepG2细胞,在治疗超声的不同声强、占空比和辐照时间作用下,观察pEGFP-N3质粒在HepG2细胞中的表达。荧光显微镜下观察绿色荧光蛋白在HepG2细胞中的表达,流式细胞仪检测细胞的转染率,MTT法检测细胞活性。结果在超声声强为2 W/cm2、占空比为20%、照射时间为60 s时,HepG2细胞的转染率最高,达到(11.53±2.15)%,且细胞存活率>85%。结论 UTMD是一种有效的基因转染方法,不同的超声转染参数对细胞活力和基因传输效率有较大影响,对其进行优化后可减少细胞损伤,增强基因转染。更多还原

【Abstract】 Objective To investigate the transfection efficiency and safety,and the optimal conditions of delivering plasmid pEGFP-N3 to HepG2 by ultrasound targeted microbubble destruction(UTMD). Methods HepG2 cells were transfected with the plasmid pEGFP-N3 under different therapeutic ultrasound(TUS) parameters(different intensities,duty cycles,exposure times).The transfection efficiency was assessed by fluorescent microscopy and flow cytometry.The survival rate was measured with MTT. Results An exposure time of 60 s at 2 W/cm2 and 20% duty cycles resulted in the optimal efficacy(11.53±2.15)% on delivering the plasmid pEGFP-N3 into the HepG2 cells,with over 85% of the transfected cells viable. Conclusion UTMD is an effective method of gene delivery,with optimal condition it enhances transfection efficiency without significant cell death.更多还原