目的 探讨超声联合微泡造影剂介导NET-1 siRNA在人肝癌细胞中的转染效果.方法 将培养的HepG2细胞分为5组:A组为空白对照；B组培养瓶中仅加入NET-1 siRNA；C组为加入脂质体及NET-1 siRNA;D组为加入造影剂及NET-1 siRNA并给予超声辐照；E组为加入脂质体、造影剂、NET-1 siRNA,并给予超声辐照.之后以荧光显微镜检测NET-1 siRNA转染率,RT-PCR法检测各组细胞NET-1 mRNA的基因表达,Western-Blotting法检测各组细胞NET-1表达情况,MTT检测HepG2细胞生长情况.结果 与C组、D组相比,E组NET-1 siRNA转染率明显提高(P＜0.05)；D组和E组细胞内NET-1 mRNA及NET-1蛋白的表达抑制率均高于其他各组(P均＜0.01),而转染后24～72 hHepG2细胞增殖受到明显抑制.结论 超声联合微泡造影剂能有效促进NET-1 siRNA在肝癌细胞中的表达,并抑制肝癌细胞的增殖.%Objective To investigate the effect of NET-1 siRNA transfected into HepG2 cells by microbubbles contrast a-gents combined with ultrasound exposure. Methods HepG2 cells were divided into 5 groups, i. e. control group (Group A), NET-1 siRNA group (Group B), Lipofectamine+NET-1 siRNA group (Group C), SonoVue+NET-1 siRNA group (Group D) and Lipofectamine+NET-1 siRNA+SonoVue group (Group E). The latter two groups were irradiated by ultrasound. Then the transfection rates of siRNA were observed using fluorescence microscope. RT-PCR and Western-Blotting were used to evaluate the expression of NET-1 mRNA and NET-1 protein. The proliferation of HepG2 cells was observed by MTT assay. Results The NET-1 siRNA transfection rate of Group E was higher than that of Group C and Grops D (P<0. 05) , the protein expression inhibition rate of NET-1 mRNA and NET-1 protein of group D and E were higher than those of the other groups (P<0. 01) , while the proliferation of cells were significantly inhibited 24 to 72 h after transfection. Conclusion The method of ultrasound contrast agents combined with ultrasound exposure can increase the transfection effect of NET-1 siRNA and inhibit the proliferation of HepG2 cells.
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