超声靶向微泡破碎介导EGFP质粒转染肝癌细胞的研究

Ultrasound targeted microbubble destruction enhances gene transfection into HepG2 cells in vitro

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。

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 eytometry. The survival rate was measured with MTT. Results An exposure time of 60 s at 2 W/era2 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.