乙肝病毒衣壳作为肝癌基因转移载体对肝癌是否有靶向性

Whether the hepatitis B virus envelope as a gene transfer vector has targeting ability for liver cancer

背景与目的：乙型肝炎病毒（HBV）具有嗜肝性的主要原因在于病毒衣壳（HBVE）上镶嵌的外衣壳蛋白PreS1,利用此生物特性有可能将HBVE改造成肝癌靶向性基因转移载体.本研究探讨乙型肝炎病毒衣壳作为基因转移载体对肝癌是否有靶向性.方法：采用PEG 8000病毒浓缩法、β-丙内脂法从Hep G 2.2.15细胞上清液中制备乙型肝炎病毒衣壳（HBVE）,用它包裹绿色荧光蛋白质粒（pIRS2-EGFP）后得到基因转移载体复合物HBVE-GFP,用HBVE-GFP分别转染Hep G2、A549、HeLa、FB细胞,通过荧光显微镜观察绿色荧光蛋白表达情况,通过流式细胞仪检测细胞发光率.结果：荧光显微镜下观察可见,各组均可见绿色荧光蛋白的表达,Hep G2组较之其他3组具有更高的荧光强度;流式细胞仪检测结果显示,Hep G2组的转染效率为（71.35±0.03）%,显著高于其他3组（P＜0.001）,且其荧光强度是其他3组的3～4倍（P＜0.001）.结论：乙型肝炎病毒衣壳作为载体对肝癌具有较好的靶向性.

Background and purpose： The reason for hepatitis B virus （HBV） with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope （ HBVE）. So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods： HBVE was obtained from the superuatant of Hep G 2.2.15 cells through PEG8000 system and β-propiolactone method. The pIRS2 -EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein （GFP） expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer. Results： The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was （ 71.35 ± 0. 03） % , much higher than other groups（ P 〈0.001） and the fluorescent intensity of the HepG2 group was also 3-4 times higher than the other 3 groups （P 〈0.001）. Conclusions： The hepatitis B virus envelope （HBVE） has a good targeting ability for liver cancer cells.