比较不同频率低频超声联合微泡促进脂质体介导的pEGFP质粒转染人前列腺癌细胞的实验研究  

A comparison study of different low-frequency ultrasound combining with microbubbles promote the liposome-mediated pEGFP plasmid transfection into human prostate cancer PC3 cells

作　　者：张蔚[1] 白文坤[1] 寿文德[1] 王玉[1] 陈旖旎[1] 杨雨[1] 胡兵[1] 

机构地区：[1]上海交通大学附属第六人民医院超声医学科 上海超声医学研究所,上海市200233

出　　处：《临床超声医学杂志》2015年第3期145-148,共4页Journal of Clinical Ultrasound in Medicine

基　　金：国家自然科学基金委面上项目(81271597);国家自然科学基金青年项目(81401421)

摘　　要：目的比较不同频率低频超声联合微泡促进脂质体介导的p EGFP质粒转染人前列腺癌PC3细胞。方法实验共分7组:空白对照组仅人前列腺癌PC3细胞株,不进行任何处理;质粒组每毫升细胞悬液中加入1μg质粒;脂质体组每毫升细胞悬液中加入100μl转染液;脂质体+微泡组每毫升细胞中悬液加入100μl转染液和200μl微泡,低频超声联合脂质体+微泡组每毫升细胞悬液中加入100μl转染液和200μl微泡,同时采用声功率为400 m W/cm2的脉冲超声波辐照模式,辐照时间240 s,占空比设为1∶1,依据辐照频率不同又分3个亚组,分别为20 k Hz超声组、500 k Hz超声组和1 MHz超声组。每组设6个复孔。各组经处理后继续培养24 h,荧光显微镜观察转染情况;流式细胞仪检测各组转染率。结果荧光显微镜下,低频超声联合脂质体+微泡组PC3细胞胞质内可见大量绿色荧光蛋白表达,明显多于其他各组,各亚组间又以20 k Hz超声组绿色荧光蛋白较多;脂质体组和脂质体+微泡组绿色荧光蛋白表达量亦多于空白对照组和质粒组。流式细胞仪检测显示,低频超声联合脂质体微泡组转染率高于质粒组,其中20 k Hz超声组转染率最高,明显高于其他各组,差异有统计学意义(P<0.05)。脂质体组与脂质体+微泡组之间、500 k Hz超声组与1 MHz超声组之间转染率比较差异无统计学意义。结论低频超声辐照微泡可显著促进脂质体介导的p EGFP质粒转染人前列腺癌PC3细胞。在相同声功率、相同辐照面积下,随着辐照频率的升高,转染率呈现下降趋势。Objective To compare the liposome-mediated pEGFP plasmid transfection into human prostate cancer PC3 cells by combining different low-frequency ultrasound with microbubbles. Methods Ultrasound with acoustic power of 400 mW/cm^2 and impulsive wave mode was used. The PC3 cells were divided into seven groups,including control group（A ）, plasmid group （B）, liposome group（C ）,liposome and microbubble group（D）, and ultrasound combined with liposome and microbubhle group （E）. Group E was further divided into three sub-group：20 kHz group（El）,500 kHz group（E2） and 1 MHz group（E3）. After treatment,the cell suspensions were cultured for 24 h. Fluorescence microscopy and flow eytometry were used to observe the transfection situation and detect transfection efficiency. Results Under a fluorescence microscopy, group E showed more green fluorescence protein expression than other groups, and group E1 showed more green fluorescence protein expression than group E2 and group E3.Group C and group D were detected with more green fluorescence protein expression than group A and group B. Flow cytometry revealed the transfection efficiency of group E was significantly higher than that of group C,group E1 had the highest transfection efficiency in all the group（P〈0.05 ）.There was no significant difference of transfection efficiency between group C and group D,group E2 and group E3.Conclusion Low-frequency ultrasound combined with microbubbles can significantly promote the transfection of liposome-mediated pEGFP plasmid into human prostate cancer PC3 ceils. The transfection efficiency tends to decline with the increasing of frequency if the acoustic power is defined.

关 键 词：超声检查 低频 微泡 质粒转染 前列腺癌细胞 

分 类 号：R445.1[医药卫生—影像医学与核医学]