摘要
目的探讨不同超声辐照时间和微泡剂量与基因转染效率之间的关系，初步摸索适宜转染的参数条件。方法质粒DNA用增强型绿色荧光蛋白质粒（pEGFP），以人肝癌细胞（HepG2）为研究对象。超声频率1MHz，声强0．5w／cm^2，对加入pEGFP和不同剂量微泡的HepG2细胞辐照不同的时间，24h后用荧光显微镜观察绿色荧光蛋白表达情况，流式细胞仪检测基因转染率，台盼蓝染色检测细胞存活率。结果各实验组均可见不同程度荧光蛋白表达，辐照时间为20S和微泡剂量为60μl时可达到较理想的转染效率。结论在超声频率和声强一定的条件下，不同超声辐照时间和微泡剂量有不同的基因转染效率，20S、60μl是一个较理想的参数，为基因治疗进一步研究提供了参考。
Objective To investigate the relationship of gene transfection efficiency with different ultrasound exposure time and different dose of microbubble,and to find the appropriate ultrasound parameters for gene transfection. Methods Plasmid encoding enhanced green fluorescent protein（pEGFP） was chosen as a report gene and HepG2 cells were chosen as research object. The HepG2 cells plus pEGFP and different dose of microbubble were exposed to ultrasound（1 MHz,0.5 W/cm^2） with varying time. Twenty-four hours later, the expression of EGFP in the cells was observed by fluorescence microscope,the transfection efficiency was assessed by FACS and the cell viability was observed by trypan blue exclusion. Results The expression of EGFP in all experimental groups was different,and the approving transfeetion efficiency was got by ultrasound exposed for 20 s when the dose of microbubble was 60 μ1. Conclusions With fixed ultrasound frequency and power, different transfection efficiency was got when the exposure time and dose of microbubble were different. The appropriate parameter was 20 s,60 μ1, which can supply information for further study.
出处
《中华超声影像学杂志》
CSCD
北大核心
2009年第9期805-808,共4页
Chinese Journal of Ultrasonography
基金
湖北省自然科学基金（2008CDB148）
关键词
超声检查
微气泡
转染
细胞系
肿瘤
Ultrasonography
Microbubbles
Transfection
Cell line, tumor
