微泡造影剂联合超声辐照介导的绿色荧光蛋白质粒转染小鼠肝癌的实验研究

目的探讨微泡造影剂SonoVue联合超声辐照在介导体内基因转染中的作用。方法建立小鼠肝癌皮下移植瘤模型，尾静脉注入绿色荧光蛋白质粒（pEGFP），添加或不添加SonoVue，脉冲多普勒超声辐照（1MHz，2W／cm^2）瘤组织。持续时间1、5、10min，7d后流式细胞仪、荧光显微镜评价pEGFP转染率。HE染色行肿瘤病理学检查。结果SonoVue联合超声辐照组pEGFP的转染率显著高于单纯超声辐照组（P〈0．01）；仅SonoVue与单纯pEGFP2组间转染率无显著差异（P〉0．05）；辐照时间5、10min时pEGFP表达明显高于1min（P〈0．05）。5与10min组间pEGFP表达无显著差异（P〉0．05）。HE染色肿瘤组织无坏死灶出现。结论微泡造影剂联合超声辐照可明显提高基因转染率，且对组织无损害。

Effect of Microbubble-enhanced Ultrasound Exposure on Gene Transfection of Green Fluorescent Protein Plasmid in Hepatocellular Carcinoma:Experimental Study

Objective To investigate the effect of microbubble contrast agent plus ultrasound exposure on gene transfection in vivo.Methods The model of hepatic carcinoma in mice were established by injecting hepa1-6 cells subcutaneously.Plasmid enhanced green fluorescent protein (pEGFP) was injected into the tail vein with or without SonoVue,Pulsed Doppler ultrasound was used (1 MHz,2 W/cm~2)for 1,5 and 10 minutes.The expression of EGFP in hepatocarcinoma cell was examined by flow cytometry and fluorescence microscopy,and the tumor tissue was observed with HE stain.Results The transfection efficiency of pEGFP was significantly higher in group ultrasound combined with SonoVue than that in group ultrasound exposed only(P<0.01).The expression of pEGFP exposed for 5 and 10 min were increased significantly compared with that exposed for 1 min (P<0.05),whereas there was no statistical difference between the group of 5 and 10 min.There was no obvious necrosis of tumor tissue in every group.Conclusions Microbubble contrast agent plus ultrasound exposure can significantly improve the transfection efficiency of pEGFP in tumors without obvious damage to the exposed tissue.