超声微泡造影剂介导mir-199a转染人肝癌HepG2细胞的实验研究  

Experimental study on miRNA-199a transfected into human hepatoma HepG2 cells mediated by microbubble ultrasound contrast agent

作　　者：过新民[1] 阮黎[2] 全显跃[1] 

机构地区：[1]南方医科大学珠江医院,广州市510280 [2]暨南大学第四附属医院,广州市红十字会医院,510220

出　　处：《实用医学杂志》2013年第24期3978-3982,共5页The Journal of Practical Medicine

基　　金：2012年度广东省科技计划项目立项(编号:20120318079)

摘　　要：目的:探讨微泡造影剂介导miRNA-199a在人肝癌HepG2细胞中的效率和稳定性。方法:利用超声造影剂六氟化硫微泡(SF6)介导miRNA-199a在人肝癌HepG2细胞中的转染率。将待转染的细胞分为以下5组:质粒组;质粒+超声组(4亚组);质粒+超声+SF6组;质粒+脂质体组;空白对照组。通过MTT比色鉴定微泡造影剂对细胞的影响程度。结果:SF6浓度<10%时,微泡均不会引起明显的细胞死亡(细胞存活率>80%),当微泡造影剂浓度达20%时,对细胞的杀伤力明显增强(细胞存活率<65%);通过4组平行因素[机械指数(MI)分别为0.12、0.20、0.28、0.35]的超声微泡造影剂转染实验,利用荧光显微镜与流式细胞术发现造影剂的最适宜的浓度(10%)下,超声频率为2.0 MHz,MI:0.28时,转染效率可达到(26.31±0.72)%。结论:在一定辐照条件下,微泡浓度10%介导miRNA-199a转染肝癌HepG2细胞效率较高,且显著高于传统的脂质体转染。Objective To explore the efficiency and stability of the transfection of miRNA-199a into human hepatoma HepG2 cells mediated by microbubble ultrasound contrast agent. Methods Microbubble contrast agent （SF6） was used to transfect miRNA-199a into human hepatoma HepG2 cells. HepG2 cells were divided into 5 groups： plasmid group, plasmid ＋ ultrasound group （divided into 4 sub-groups）, plasmid ＋ ultrasound ＋ SF6 group, plasmid ＋ liposome group, and blank control group. MTT staining was used to assay the lethality of microbubble contrast agent to cells. Results When the concentration of SF6 was less than 10%, there was no significant cell death （cell viability 〉 80%）; when that was more than 20%, there was significant lethality to cells （cell survival 〈 65%）. Four sets of parallel factors （MI： 0.12, 0.20, 0.28, 0.35） were set for the microbubble transfection experiments, and at the optimal microbubble concentration （10%） and at the optimal sound intensity （2.0 MHz, MI 0.28）, the transfection efficiency of （26.31 ± 0.72）% could be achieved by fluorescence microscopy and flow cytometry. Conclusion In certain irradiation conditions, the cell transfection was effective when used 10% microbubble ultrasound contrast agent to transfect miRNA-199a into human hepatoma HepG2 cells, and the transfection efficiency was significantly higher than that of conventional liposomal transfection.

关 键 词：HEPG2细胞 mir-199a 六氟化硫 超声微泡 细胞转染 

分 类 号：R735.7[医药卫生—肿瘤]