在不同细胞系中共聚物对超声介导基因转染的影响  

The function of pluronic block copolymers in ultrasound-mediated gene delivery in different cell lines

作　　者：陈云超[1] 黄道中[1] 李开艳[1] 王志辉[1] 洪恺[1] 王芬[1] 张青萍[1] 

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

出　　处：《中华超声影像学杂志》2008年第2期163-166,共4页Chinese Journal of Ultrasonography

摘　　要：目的 探讨超声和共聚物在不同细胞系中对基因转染的协同作用。方法 应用三种水溶性不同的共聚物F127，L61和P85；超声频率为1MHz，脉冲重复频率100Hz，占空系数为20％，用1W／cm^2声强照射20s。选用C2C12，3T3-MDEI，CHO和H2K四种细胞系为研究对象。质粒DNA为可产生绿色荧光蛋白的GFP。应用荧光显微镜和流式细胞仪评价转染率，台盼蓝染色评价细胞的成活率。结果 超声可介导所有四种细胞系的基因转染。在C2C12，3T3-MDEI和CH0细胞系，共聚物可显著促进超声介导的基因转染效率（P〈0．01），对H2K细胞系共聚物不但不能促进超声介导的基因转染，其中疏水性共聚物P85和L61反而使超声介导的基因转染效率降低（P〈0．05）。结论 共聚物对不同细胞系超声介导的基因转染效率产生不同的影响，这种差异可能与细胞膜蛋白缺陷有关。Objective To explore whether the ultrasound exposure and pluronics can induce gene delivery synergistically in different cell lines or not. Methods Three different pluronics,F127,L61 and P85 were investigated for their effects on ultrasound-mediated gene transfection in four cell lines （C2C12,3T3-MDEI,CHO and H2K）. The frequency of the ultrasound system was 1 MHz and the pulse repetition frequency was 100 MHz with fixed 20% duty cycle and the ultrasound intensity of 1 W/cm^2 with the duration of 20 s was used in the experiment. Plasmid encoding green fluorescent protein（GFP） was used as a reporter. Transfection rates and cell viability were assessed by fluorescence microscopy, flow cytometer （FACS） and trypan blue exclusion. Results Ultrasound could induce gene delivery in all four cell lines. The three pluronics can enhance the ultrasound- mediated gene transfection rate in C2C12,3T3-MDEI and CHO cell lines（ P〈0. 01）,but not in H2K cell line. On the contrary,the hydrophobic pluronics of P85 and L61 decreased the gene transfection rate induced by ultrasound （P〈 0.05）. Conclusions Pluronics shows different effects on ultrasound-mediated gene delivery in different cell lines. The difference may be induced by the defect some of the membrane protein in the H2K cell line.

关 键 词：超声检查 转染 细胞系 转化 共聚物 

分 类 号：R346[医药卫生—基础医学]