作为基因转移载体的乙型肝炎病毒衣壳对肝癌细胞的靶向性

Targeting efficiency of hepatitis B virus envelope as a gene transfer vector on liver cancer cells

目的:探讨乙型肝炎病毒衣壳作为基因转移载体对肝癌细胞的靶向性和有效性。方法:采用PEG8000病毒浓缩法、β-丙内脂法从能持续产生HBV的HepG2.2.15细胞上清液中制备乙型肝炎病毒衣壳(hepatitisBvirusenvelope,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,用脂质体包裹pIRS2-EGFP后得到基因转移载体复合物Liposome-GFP。用HBVE-GFP及作为对照的Liposome-GFP转染人成肝细胞瘤HepG2细胞来研究其转导效率;用HBVE-GFP分别转染HepG2细胞及作为对照的人肺腺癌细胞A549、人宫颈癌细胞HeLa、人皮肤成纤维细胞FB来研究其靶向性;荧光显微镜观察绿色荧光蛋白表达情况,流式细胞术检测细胞发光率。结果:(1)有效性检测:Liposome-GFP组及HBVE-GFP组均可见绿色荧光蛋白的表达,HBVE-GFP组的荧光强度较Liposome-GFP组更强(P<0.01);Liposome-GFP组的转染效率为(49.97±2.37)%,而HBVE-GFP组为(70.65±3.15)%,两者差异显著(P<0.01)。(2)靶向性检测:各种细胞均可见绿色荧光蛋白的表达,HepG2细胞较其他细胞具有更强的荧光强度(P<0.01),HepG2细胞的转染效率为(71.35±0·03)%,显著高于其他3组(P<0.01)。结论:乙型肝炎病毒衣壳作为基因转移载体对肝癌细胞具有较好的转染有效性和靶向性。

Objective： To observe the transfection efficacy and targeting efficiency of hepatitis B virus envelope （HBVE） as a gene transfer vector for liver cancer cells, Methods： HBVE was obtained from the supernatant of HepG2. 2. 15 cells with a PEG8000 system and β-propiolactone, The pIRS2-EGFP was packed with HBVE to obtain HBVE-GFP and was packed with liposome to obtain Liposome-GFP, HBVE-GFP and Liposome-GFP were used to transfect human hepatoblastoma cell line HepG 2 to study the transfection efficiency, HepG 2, A 549, HeLa and FB cells were transfected with HBVE-GFP to appraise the targeting ability of HBVE-GFP. GFP protein expression was observed under a fluorescent microscope and the ratio of GFP positive cells was determined by flow cytometry, Results： （ 1 ） Transfection efficiency： The GFP protein was observed in both the liposome group and the HBVE group under the fluorescent microscope; the fluorescent intensity in the HBVE group was 3-4 times that of liposome group as determined by flow cytometry （ P 〈 0. 01 ）. The transfection rate of liposome group was （49.97 ± 2. 37 ） % and was （ 70.65 ± 3.15 ） % in HBVE group. （2） Targeting ability ： The GFP was observed in all the 4 types of cells under the fluorescent microscope ; the fluorescent intensity of the HBVE group was 2-3 times those of the other 3 groups （P 〈0.01 ）. The transfection rate of HepG2 group was （71. 35 ±0.03 ） % , significantly higher than those of the other 3 groups （ P 〈 0.01 ）. Conclusion： HBVE as a gene transfer vector has a satisfactory transfection efficacy and targeting ability for liver cancer cells.