不同浓度超声微泡结合共聚物P85对人肝癌细胞HepG2基因转染的影响  

Impact of different microbubble concentration and pluronic P85 under ultrasound irradiation mediated gene transfection to HepG2 cell

作　　者：张喜君[1] 李开艳[1] 崔贤[1] 胡良军[1] 陈云超[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院超声影像科,湖北武汉430030

出　　处：《中国医学影像技术》2011年第4期669-672,共4页Chinese Journal of Medical Imaging Technology

基　　金：湖北省自然科学基金(2008CDB148);国家自然科学基金(30970882)

摘　　要：目的不同浓度超声微泡造影剂(MB)结合共聚物P85后联合一定强度超声(US)辐照,初步摸索最适人肝癌细胞HepG2质粒转染及表达的MB浓度。方法以人肝癌细胞HepG2为研究对象,以绿色荧光蛋白(EGFP)质粒为报告基因,添加共聚物P85和不同浓度的MB后进行脉冲多普勒US辐照。24 h后台盼蓝染色评价细胞存活率,荧光显微镜和流式细胞仪评价细胞的基因转染率。结果 MB浓度在30%时达到较理想的转染率(22.14±3.06)%,且细胞存活率无明显下降(55.73±3.32)%。添加MB后基因转染率均较pEGFP+P85+US转染率高。结论 MB与共聚物P85结合同时给予US辐照可增强基因转染率,且在MB浓度为30%时达到较理想的转染率。Objective To investigate the appropriate microbubble（MB） concentration with pluronic P85 that can enhance the gene transfection and expression to HepG2 cell under ultrasound（US） irradiation.Methods Plasmid encoding enhanced green fluorescent protein（pEGFP） was used as a report gene.HepG2 cell and plasmids DNA with P85 and different microbubble concentration were exposed to US,and after 24 h the transfection rates and cell viability were assessed with fluorescence microscopy,FACS and trypan blue exclusion.Results The approving transfection efficiency was got at 30% microbubble concentration,with ideal gene transfection efficiency（22.14±3.06）% and without obvious decreasing of survival rate（55.73±3.32）%.Higher transfection rate than pEGFP＋US＋P85 could be obtained through adding different concentration of microbubble.Conclusion Microbubble combined with pluronic P85 can enhance the gene transfection under ultrasound irradiation,and the approving transfection rate can be obtained at 30% microbubble concentration.

关 键 词：共聚物 微泡 超声检查 基因转染 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R445.1[医药卫生—基础医学]