小分子聚乙烯亚胺与腺病毒结合增强基因转染效率的研究

Combination of low molecular weight polyehylenimine with adenovirus enhances gene transduction efficiency

目的提高重组腺病毒在细胞中,特别是缺乏柯萨奇腺病毒受体的细胞中的转染效率。方法将阳离子材料聚乙烯亚胺PEI与重组腺病毒通过电荷作用形成复合物,用激光粒度仪及电位分析仪测定其粒径和电位;以表达β半乳糖苷酶的重组腺病毒作为模型病毒,研究其在富含(A549)或者缺乏(MDCK、LLC)柯萨奇腺病毒受体细胞中的转染效率和毒性,并用荧光标记复合物,观察其进入细胞的能力。结果与聚乙烯亚胺结合形成复合物可明显促进腺病毒进入细胞的能力,从而提高腺病毒在各种细胞中的转染效率,同时发现小分子的PEI-2 kD由于其较好的生物相容性,对腺病毒进入细胞后的传递影响较小,所以最终的基因传递效率要显著优于PEI-25 kD;PEI-2 kD在本复合物中表现出的细胞毒性要小于PEI-25 kD。结论PEI-2 kD可以用于病毒载体和非病毒载体结合的基因传递研究。

OBJECTIVE To enhance the transduction efficiency of adenovirus on cells,especially on coxsackievirus and adenovirus receptor（CAR） deficient cells.METHODS Adenovirus was combined with cationic material,polyethylenimine（PEI） through charge interaction to form complexes as new gene vector.The particle size and Zeta potential of the complexes were measured by photon correlation spectroscopy.And the transduction efficiency and cytotoxicity were analyzed on CAR overexpressed（A549） or down-regulated（MDCK） cells by using recombinant adenovirus expressing β-galactosidase.The fluorescence labeled complexes were also tested for their cell enter ability.RESULTS Combination with PEI to form complexes could significantly enhance cell uptake of adenovirus,thus increased the gene transfer efficiency.Additionally,we found that PEI-2 kD as a low molecular PEI incorporated in Ad/PEI,had less cytotoxicity and higher biocompatibility,more importantly,less effect on the intracellular transfer of adenovirus,like endosome escape and nuclear localization,and subsequently higher gene transfer compared with PEI-25 kD.CONCLUSION PEI-2 kD is a useful material in the combination of viral and non-viral gene delivery strategy.