超声波与微泡声学造影剂增强体外培养肿瘤细胞基因转染实验研究  

Experimental study of enhanced gene delivery to cultured tumor cells in vitro by ultrasound and contrast agent

作　　者：冉海涛[1] 任红[2] 王志刚[1] 郑元义[1] 张群霞[1] 许川山[1] 

机构地区：[1]重庆医科大学超声影像学研究所,重庆400010 [2]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《中国医学影像技术》2005年第8期1151-1154,共4页Chinese Journal of Medical Imaging Technology

基　　金：国家自然科学基金重点项目资助(30430230)。

摘　　要：目的观察超声波与微泡声学造影剂能否增强体外培养肿瘤细胞基因转染及其转染效率,初步摸索适宜的超声辐照条件。方法将培养的HepG2细胞分为3组,第1组为单纯造影剂转染组:加入5μg绿色荧光蛋白质粒(PEGFP-C1)与微泡声学造影剂混合液进行转染;第2组为脂质体转染组:用相同浓度PEGFP-C1质粒进行常规脂质体转染;第3组为造影剂加超声辐照转染组:加入5μgPEGFP-C1质粒与微泡声学造影剂混合液,然后分别用0.25W/cm2、0.5W/cm2、1.0W/cm2超声波经培养板底部辐照10s。24h后用荧光显微镜观察各转染组细胞荧光蛋白表达情况,计录每高倍视野(400×)荧光蛋白表达阳性细胞数。结果单纯造影剂转染组未见荧光蛋白表达阳性细胞,造影剂加超声辐照转染组可见不同程度荧光蛋白表达,其中以0.5W/cm2超声辐照组表达最强,每高倍视野绿色荧光蛋白表达阳性细胞数为(20.7±3.2)个/HP,高于0.25W/cm2与1.0W/cm2超声辐照组[分别为(4.8±1.9)个/HP;(9.9±2.3)个/HP],与脂质体转染组[(22.1±3.5)个/HP]比较无显著差异(P>0.05)。结论微泡声学造影剂加一定强度的超声辐照能显著增强体外培养肿瘤细胞的基因转染与表达,超声辐照条件是影响转染率的重要因素,适宜条件时其转染效率与常规脂质体转染方法无显著差异。Objective To investigate whether ultrasound mediated microbubbles destruction could enhance the gene transfection in cultured tumor cells in vitro. Methods Cultured HepG2 cells were devided into three groups. Group 1 was taken as simple microbubble transfection group： a mixture of microbubbles and p^EGFP plasmid （5 μg） was added to the cells; Group 2 was taken as liposome transfeetion group： 5 μg plasmid was used for liposome transfection as normal. Group 3 was taken as ultrasound irradiation plus microbubble group： after added a mixture of microbubbles and plasmid to the cells, ultrasound with the intensity of 0.25 W/cm^2 , 0.5 W/cm^2 , 1.0 W/cm^2 was applied under the bottom of the plate for 10 s. Twenty-four hours later, the expression of GFP in the cells was observed by fluorescence microscope, the numbers of positive expression cells in every high power field （ X 400） were counted. Results There was no positive expression cell in Group 1 ; there were different quantity of positive expression cells in Group 3. The highest GFPexpression was seen in the group in which ultrasound with intensity of 0.5 W/cm^2 was applied. The mean number of positive expression cells under each high power field were （20.7±3.2）/HP, higher than that in other subgroup （0.25 W/cm^2 or 1.0 W/cm^2） of Group 3, and there was no sig- nificant difference compared to Group 2 （P〉0.05）. Conclusion Ultrasound and microbubbles could highly enhance the gene transfection to tumor cells in vitro. The condition of ultrasound irradiation was a important factor for gene transfection. There was no significant difference between the suitable ultrasound energy and liposome transfection method.

关 键 词：超声造影剂 微泡 基因转染 

分 类 号：R445.1[医药卫生—影像医学与核医学] Q95-33[医药卫生—诊断学]