超声辐照联合PEI介导基因转染的实验研究  

Ultrasound Irradiation Accompanied with PEI Mediated Gene Transfection:in Vitro Experimental Study

作　　者：刘娜香[1] 陈云超[1] 蒋丽萍[1] 丁璐[1] 刘晓玲[1] 张青萍[1] 

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

出　　处：《中国超声医学杂志》2011年第10期865-868,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金面上项目(No.30970882)

摘　　要：目的探讨超声联合聚乙烯亚胺(PEI)在促进肝癌细胞(HepG2)的基因转染中有无协同作用。方法选用工业用和实验用PEI为研究对象。用锥虫蓝拒染法检测细胞成活率,选取合适的PEI浓度与绿色荧光蛋白基因(EGFP)质粒DNA共孵育,制备阳离子复合物(PEI/DNA)导入HepG2细胞中和/或辐照超声。24h后荧光显微镜观察绿色荧光蛋白表达,流式细胞仪定量分析转染率。结果超声可促进质粒DNA在HepG2细胞的基因转染。PEI对细胞有毒性,当PEI浓度为0.001%时细胞成活率约为70%,选此浓度用于基因转染实验。2种PEI复合物均能显著促进质粒DNA基因转染,其转染率高于单纯辐照超声介导的基因转染,差异有统计学意义(P<0.01),2种PEI的转染率工业用组(15.42±4.71)%、实验用组(12.27±3.58)%之间的差异无统计学意义(P>0.05),加入PEI联合辐照超声后转染率下降(P<0.01)。结论 PEI和超声均能促进HepG2细胞基因转染,但PEI联合辐照超声后转染率明显下降,二者没有协同作用。工业用与实验用PEI一样具有促进基因转染的作用。Objective To investigate whether ultrasound and PEI can enhance plasmid DNA transfection in HepG2 cell synergistically or not. Methods Two kinds of PEI, one used specifically in gene delivery （sPED and the other used just as general industrial materials （iPEI） , were used. The toxicity of PEI was determined by trypan blue dye exclusion. The PEIs were incubated with the plasmid DNA to prepare cationic compounds （PEI/DNA）, which were added into HepG2 cell with or without ultrasound irradiation. After 24 hours, the transfection efficiency was as- sessed by fluorescence microscope and FACS. Results Ultrasound could induce gene delivery in HepG2 cell. PEI dis- played high toxic to cell and the concentration of 0. 001 %was used in gene delivery, which induced about 70 %cell survival rate. The two PEI compounds significantly enhanced the plasmid DNA gene delivery in HepG2 cell （P〈 0.01） and the transfection rates of the iPEI and sPEI were （15.42±4.71）% and （12.27±3.58）% respectively. Even though the difference of the two PEI was not significant （P〉0.05）, their transfection rates were higher than that of the ultrasound group （P〈0.01）. On the other hand, PEI with ultrasonic irradiation group showed a lower transfec- tion rate compared to the PEI groups （P〈0.01）. Conclusions PEI and ultrasound irradiation all can promote plasmid DNA transfection in vitro respectively but they do not show synergism.

关 键 词：超声 聚乙烯亚胺 基因转染 HEPG2细胞 

分 类 号：R445.1[医药卫生—影像医学与核医学] R735.7[医药卫生—诊断学]