Department of Hospital Pharmacy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
Mol Pharm. 2011 Jun 6;8(3):913-9. doi: 10.1021/mp2001268. Epub 2011 May 18.
We developed a novel vector, electrostatically coated poly(ethylenimine) (PEI)/pDNA complexes with folic acid (FA). Without covalent binding, the FA molecules could coat the PEI/pDNA complexes, and stable anionic nanoparticles were formed at a charge ratio greater than 60. The addition of FA markedly decreased the cytotoxicity of the cationic PEI/pDNA complexes to the melanoma cell line, B16-F10 cells, which regularly expressed FA-specific receptor (FR). Furthermore, the anionic FA60/PEI/pDNA complexes showed high transgene efficiency via the FR-mediated pathway in B16-F10 cells. The FA60/PEI/pDNA complexes did not show agglutination with erythrocytes. After the intravenous injection of FA60/PEI/pDNA complexes into mice, a higher transgene efficiency than PEI/pDNA complexes was observed in the liver, kidney, spleen, and lung with FR. The gene expressions of FA60/PEI/pDNA complexes were significantly inhibited by preadministration of FA. Thus, the FA60/PEI/pDNA complexes were useful for effective gene therapy.
我们开发了一种新型载体，即带负电荷的聚（亚乙基亚胺）（PEI）/pDNA 与叶酸（FA）的复合物。在没有共价结合的情况下，FA 分子可以包裹在 PEI/pDNA 复合物上，并且在电荷比大于 60 时形成稳定的阴离子纳米颗粒。FA 的添加显著降低了阳离子 PEI/pDNA 复合物对黑色素瘤细胞系 B16-F10 细胞的细胞毒性，B16-F10 细胞通常表达 FA 特异性受体（FR）。此外，带负电荷的 FA60/PEI/pDNA 复合物通过 FR 介导的途径在 B16-F10 细胞中显示出高转染效率。FA60/PEI/pDNA 复合物与红细胞没有聚集。将 FA60/PEI/pDNA 复合物静脉注射到小鼠体内后，FR 在肝脏、肾脏、脾脏和肺中观察到比 PEI/pDNA 复合物更高的转染效率。FA 的预先给药显著抑制了 FA60/PEI/pDNA 复合物的基因表达。因此，FA60/PEI/pDNA 复合物可用于有效的基因治疗。
