乙肝病毒L颗粒作为肝癌靶向性基因治疗载体的研究  

L particles of HBV as a novel targeted vector used for gene therapy of hepatocellular carcinoma in vitro

作　　者：曾建平[1] 易继林[2] 王从俊[2] 杨志芳[1] 

机构地区：[1]解放车总医院肝胆外科,北京100853 [2]华中科技大学同济医学院附属同济医院普外科

出　　处：《中华实验外科杂志》2009年第11期1453-1455,F0003,共4页Chinese Journal of Experimental Surgery

摘　　要：目的评估乙肝病毒L蛋白颗粒作为一种新型肝癌靶向性基因治疗转运载体的可行性。方法通过电穿孔方法（电压=400V，脉冲时间=60us，pGFP：L颗粒=4：10）将绿色荧光表达质粒pGFP导入乙肝病毒L颗粒，形成的L／pGFP颗粒转染各种肝来源细胞株以及非肝来源细胞株，以脂质体作为对照转染相同细胞株，荧光显微镜检测各细胞株基因转运效率。结果L／pGFP颗粒对各种肝来源细胞株均保持较高的转染效率。对正常肝来源细胞L02的转染效率（67．0±2．6）％低于肝癌细胞株HepG2（75．0±3．5）％和7721（72．0±2．3）％，然而均显著高于脂质体的转染效率（P〈0．05）。非肝来源的乳腺癌细胞株MCF-7不能被L/pGFP颗粒有效转染。未接受电穿孔的L颗粒＋pGFP混合液不能有效转染任何细胞株。结论L颗粒能特异性、高效转运外源性基因至各种肝来源细胞，可作为一种安全高效的靶向性转运载体用于肝癌的基因治疗。Objective To evaluate the possibility of L nanoparticles of hepatitis B virus as a novel vector used for targeted gene therapy of hepatocellular carcinoma （HCC）. Methods The GFP expression plasmid was incorporated into L nanoparticles by eleetroporation （DAN：L 4：10 ,voltage 400 V ,pulse time 60 μs）. L particles with GFP plasmid and liposome were used to transfect some cell lines from liver parenchyma and breast cancer. A mixture of L particles and GFP expression plasmid with no electroporation was used as a negative control. On the 48th h after transfection, GFP fluorescence was observed under a fluorescence microscope. Results L particles with GFP plasmid transfection yielded higher transfection efficiencies in human HCC cells HepG2 （75.0 ± 3.5 ） % ,7721 （72.0 ± 2.3 ） % and normal hepatocytes （67.0 ± 2.6） % than liposome （ P 〈 0.05 ） , whereas human breast cancer MCF-7 cells did not express GFP after transfection with the L particles carrying GFP plasmid. No expression of GFP was detected in the negative controls not subjected to electroporation. Conclusion Electroporation can incorporate plasmid into L particles. L particles can effectively and specifically transfect the gene into the human HCC cells, which is availability of targeted gene therapy of HCC.

关 键 词：癌 肝细胞 基因治疗 乙型肝炎病毒 

分 类 号：R735[医药卫生—肿瘤]