超声辐照和SonoVue微泡在介导hAng-1基因体外转染时的作用和量效关系探讨  

The Role of Ultrasound Radiation and SonoVue Microbubbles in the Transfection of Gene HAng-1 in Vitro and Their Dose-effect Relationship

作　　者：周青[1] 朱丽敏[1] 陈茜[1] 郭瑞强[1] 

机构地区：[1]武汉大学人民医院超声科,武汉市430060

出　　处：《中国超声医学杂志》2009年第12期1109-1112,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金青年基金项目(No.30600141)

摘　　要：目的探讨超声辐照和SonoVue微泡分别使用和联用在介导hAng-1基因体外转染过程中的作用以及辐照强度和微泡浓度对转染效率和细胞活性的影响。方法实验分四组A组:单纯超声辐照+质粒组;B组:微泡+质粒组;C组:超声辐照+微泡+质粒组和空白对照组D组。C组内转染参数分别设置为超声照射强度0.5、1.0、1.5和2.0 W/cm^2,微泡浓度5%、10%、20%、30%和40%。将连接有eGFP-C_3-hAng-1质粒的SonoVue微泡对293T细胞进行转染,48 h后检测各组基因转染效率和细胞存活率。结果转染48 h后C组转染效率最高,荧光阳性细胞数最多,强度最大;A组转染效率很低,见少量荧光表达;B、D组无明显基因转染发生。随着超声照射强度和微泡浓度的增加,基因转染效率会逐步升高,具有统计学意义。微泡浓度大于20%、超声照射强度超过1.5 W/cm^2后基因转染效率不再升高甚至降低,细胞死亡率显著增高(P<0.01)。结论SonoVue微泡介导外源基因转染必须联合超声辐照才能获得较好的转染效率。对于hAng-1基因和SonoVue微泡,选择声强1.5 W/cm^2,微泡浓度20%是相对最佳转染条件。Objective To investigate the role of ultrasonic radiation and SonoVue microbubbles during the process of gene hAng-1 transfection in vitro and the effects of different ultrasound （US） acoustic intensity and microbubble concentration on transfection ratio and cells viability. Methods This study consisted of 4 groups., group A： US＋plasmid, group B： microbubbles＋plasmid, group C： combined US and microbubbles＋plasmid, and group D was the control group. US radiation intensity in Group C was set as 0.5, 1.0, 1.5 and 2.0 W/cm^2 and microbubbles concentration as 5%, 10%, 20%, 30% and 40%, respectively. SonoVue microbubbles linked with eGFP-C3-hAng-1 were delivered into 293T cells. 48 hours later, gene transfection ratio and cells viability under the various parameters were measured. Results 48 hours after transfection, it was group C that got the highest gene expression and the strongest intensity of fluorescence. Group A got quite low gene expression and positive fluorescence while group B and D had little gene expression. Gene expression would be significantly increased if the intensity of US and microbubble concentration were enhanced. However, if US intensity exceeded over 1.5W/cm2 and mierobubbles over 20%, gene expression would not increase significantly or could even lower down, and coupled with obviously decreased cells viability （P〈0.01） . Conclusions It is necessary of combined with ultrasound radiation when SonoVue microbubbles are utilized to deliver exogenous gene as a vector. As for gene hAng-1 and SonoVue microbubbles, US intensity 1.5W/ cm2 and 20% microbubbles were relative optimal transfection parameters.

关 键 词：基因转染 微泡 超声 辐照 

分 类 号：R445.1[医药卫生—影像医学与核医学]