自制脂质微泡联合超声辐照介导pGL3-Promoter-EGFP质粒转染卵巢癌细胞的实验研究  

pGL3-Promoter-EGFP Plasmid Transfected into SKOV3 Cells by Ultrasound-Mediated Self-made Lipid Microbubble Destruction:Experimental Study

作　　者：朱元方[1] 胡丽娜[2] 李攀[3] 王志刚[3] 

机构地区：[1]重庆医科大学附二院妇科,重庆市400010 [2]四川大学华西附二院妇科 [3]重庆医科大学附二院超声影像研究所

出　　处：《中国超声医学杂志》2008年第4期303-306,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金(No30371619);重点项目(No30430230)资助

摘　　要：目的探讨超声微泡介导基因转染卵巢癌细胞的可行性及转染效率。方法(1)以不同声强作用SKOV3细胞60s,筛选出对细胞活性无明显抑制的声强;(2)将筛选出的声强分别与不同浓度的微泡联合作用于SKOV3细胞,筛选出对细胞无明显抑制的最适声强和微泡浓度的组合;(3)将SKOV3细胞分成5组,不同方式转染pGL3-Promoter-EGFP质粒:(1)裸质粒组;(2)质粒+微泡组;(3)质粒+超声组;(4)质粒+超声+微泡组;(5)质粒+脂质体组,比较各组转染效率。结果(1)声强0.5、0.75W/cm2的超声辐照,细胞死亡率<10%;(2)0.24×108/ml或0.12×108/ml浓度的微泡联合0.5W/cm2超声辐照,细胞死亡率<10%;(3)第4组与第5组转染效率无区别(P=0.133),但高于第1、2、3组(P<0.001)。结论适当浓度微泡联合超声辐照能将外源基因在卵巢癌细胞中高效转移。Objective To investigate the feasibility and transfection efficiency of ectogenous plasmid transfected into SKOV3 cells by ultrasound-mediated microbubbles. Methods ①n order to screen out the optimal ultrasound intensity without conspicuous harm to SKOV3 cells, cells in 6 groups were exposed to different ultrasound intensity of 0. 5,0.75, 1.0, 1.25,1.5 and 2.0 W/cm^2 , respectively, and the time 60 s was continuously ;②Combined the screened ultrasound intensity with microbubbles of different concentration, SKOV3 cells were under the impact of different combinations so as to screen out the optimal ultrasound and microbubbles ; ③SKOV3 cells were divided into 5 groups and transfected with pGL3-Promoter-EGFP ： group 1, naked plasmid ; group 2, plasmid with microbubbles ; group 3, plasmid with ultrasound; group 4, plasmid with ultrasound and microbubbles; group 5 plasmid with liposome; the transfection efficiency of each group was observed. Results ①The cell death rate was〈10% ,when the ultrasound intensity was below 0. 75 W/cm^2; ② When the microbubble concentration of 0.24 × 108/ml or 0.12×108/ml was combined with the ultrasound intensity of 0.5 W/cm^2, the death rate was 〈 10%; ③The transfection efficiency of group 4 was similar to that of group 5, P= 0. 133, but were higher than that of other 3 groups, P〈0. 001. Conclusions Ectogenic plasmid can be transfected into SKOV3 cells efficiently by ultrasoundmediated microbubble destruction.

关 键 词：微泡 超声辐照 卵巢癌 转染 

分 类 号：R737.31[医药卫生—肿瘤] R454.3[医药卫生—临床医学]