Division of Clinical Pharmacology, 2nd Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
Int J Pharm. 2012 Jan 17;422(1-2):510-5. doi: 10.1016/j.ijpharm.2011.11.024. Epub 2011 Nov 24.
Transfection efficiency of non-viral gene vectors is influenced by many factors, including chemical makeup, cellular uptake pathway and intracellular delivery. To investigate the effect of lipid saturation on transfection efficiency of polycation liposomes (PCLs), a soybean phospholipids (SPL), egg phospholipids (EPL) and hydrogenated soybean phosphatidylcholine (HSPC) series was used to prepare PCLs. Testing these PCLs in a luciferase assay indicated that with increasing saturation (SPL<EPL<HSPC), the ability to confer gene expression decreased. The effect of protamine combined with these PCLs was also studied in different cell lines. Improved transfection because of protamine incorporation was dependent on lipid saturation and on the cell line tested. The kinetics of cellular uptake and intracellular distribution was studied using flow cytometry and laser scanning confocal microscope, which showed that naked oligonucleotide (ODN) and PCLs/ODN complexes became equilibrium after 4h incubation. PCLs containing SPL (PCLs-S) and 1,2-dieleoyl-sn-glycero-3-phosphoethanolamine (PCLs-D) increased uptake rates by 2.20- and 5.45-fold, respectively. Furthermore, pCMV-IL-12 transfection mediated by PCLs-D showed excellent tumor inhibition efficiency compared with control and naked pCMV-IL-12 treatments in vivo.
非病毒基因载体的转染效率受多种因素影响，包括化学组成、细胞摄取途径和细胞内传递。为了研究脂质饱和度对阳离子脂质体（PCL）转染效率的影响，我们使用大豆磷脂（SPL）、卵磷酯（EPL）和氢化大豆磷脂酰胆碱（HSPC）系列制备了 PCL。在荧光素酶检测实验中测试这些 PCL，结果表明随着饱和度的增加（SPL<EPL<HSPC），赋予基因表达的能力降低。我们还研究了鱼精蛋白与这些 PCL 的组合在不同细胞系中的作用。由于鱼精蛋白的掺入而改善的转染效率取决于脂质饱和度和所测试的细胞系。我们使用流式细胞术和激光扫描共聚焦显微镜研究了细胞摄取和细胞内分布的动力学，结果表明裸寡核苷酸（ODN）和 PCLs/ODN 复合物在孵育 4 小时后达到平衡。含有 SPL（PCLs-S）和 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺（PCLs-D）的 PCL 分别将摄取率提高了 2.20 倍和 5.45 倍。此外，与对照和裸 pCMV-IL-12 处理相比，PCLs-D 介导的 pCMV-IL-12 转染在体内显示出优异的肿瘤抑制效率。
