超声靶向微泡破裂介导EGFP质粒转染肝癌细胞的研究  

The Research of EGFP Plasmid Transfection in Hepatoma Cells Mediated by Ultrasound Targeted Microbubble Destruction

作　　者：刘同刚[1] 沙凯辉[2] 李云华[1] 张立国[1] 刘贤贤[1] 

机构地区：[1]滨州医学院附属医院,滨州256603 [2]滨州医学院护理学院

出　　处：《肝胆外科杂志》2012年第3期215-217,共3页Journal of Hepatobiliary Surgery

摘　　要：目的探讨超声靶向微泡破裂(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,在不同治疗超声的声强、占空比和辐照时间作用下,观察pEGFP-N3质粒在HepG2细胞中的转染。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7转染率最高,达到11.53%±2.15%,且细胞生存率大于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 Investigate the effectiveness and safety of EGFP plasmid transfected HepG2 cells mediated ultrasound targeted microbubble destruction（UTMD） and Optimize the ultrasound parameters. Methods Culture the HepG2 cells in vitro. On the different of sound intensity of treating ultrasound, the duty cycle and the irradiation time, observe the HepG2 cells transfected by the pEGFP-N3 plasmid. Under the fluorescence microscope, observe the HepG2 cells expressing the green fluorescent protein, detect the transfection rate in the flow cytometry, assay the cell activity by MTT. Results When the ultrasonic intensity is 2w/cm2, the duty cy- cle is 20% , and the exposure time is 60s, HepG2 cells have the highest transfected rate reaching 11.53% ± 2. 15% , and the cell survival rate is greater than 85%. Conclusion UTMD is an effective method of gene transfection, on the cell viability and gene trans- fer efficiency, the different parameters of ultrasound transfection have a large impact, when they are optimized,it can reduce the damage on the cells, improve the rate of gene transfection.

关 键 词：超声靶向微泡破裂 肝癌细胞 基因转染 

分 类 号：R735.7[医药卫生—肿瘤]