Sun Jingxin, Zhou Yuanfei, Jin Guangyu, Jin Yong, Quan Jishan
Department of Pharmaceutics, College of Pharmacy, Yanbian University, Yanji 133000, China.
JS and YZ contributed equally to this work.
Iran J Pharm Res. 2021 Fall;20(4):265-277. doi: 10.22037/ijpr.2021.115099.15219.
In order to improve the transfection efficiency of gene vectors and monitor the effect of gene therapy, this study prepared a drug delivery vector with dual functions. The thiourea reaction was used to synthesize polyethyleneimine (PEI, MW: 1.8 kDa) with superparamagnetic iron oxide nanoparticles (SPION) (PEI1800-SPION), and the lipid polycationic gene vector PEI1800-SPION loaded cationic liposome (LP-PEI1800-SPION) was further prepared by ethanol injection method. Agarose gel electrophoresis experiment, cytotoxicity experiment, and gene silencing experiment were used to evaluate the vector and screen the optimal prescription of LP-PEI1800-SPION/siRNA. When the weight ratio of LP-PEI1800-SPION to siRNA is 20, the transfection efficiency of the nanoparticles was the highest, and the silencing efficiency of the target protein was the largest. The cytotoxicity of LP-PEI1800-SPION was low; when the concentration was in the range of 1-50 μg/mL, the survival rate of the four types of cells was above 80%. Prussian blue staining experiments and MRI imaging experiments showed that cells had significant uptake and imaging capabilities for LP-PEI1800-SPION. In conclusion, the visualized polycationic lipid siRNA delivery vehicle (LP-PEI1800-SPION) was successfully prepared in this experiment, which provides a research basis for further theranostics of liver cancer.
为了提高基因载体的转染效率并监测基因治疗效果,本研究制备了一种具有双重功能的药物递送载体。利用硫脲反应将超顺磁性氧化铁纳米颗粒(SPION)与聚乙烯亚胺(PEI,分子量:1.8 kDa)合成(PEI1800-SPION),并通过乙醇注入法进一步制备负载阳离子脂质体的脂质聚阳离子基因载体PEI1800-SPION(LP-PEI1800-SPION)。采用琼脂糖凝胶电泳实验、细胞毒性实验和基因沉默实验对载体进行评价,并筛选LP-PEI1800-SPION/siRNA的最佳处方。当LP-PEI1800-SPION与siRNA的重量比为20时,纳米颗粒的转染效率最高,目标蛋白的沉默效率最大。LP-PEI1800-SPION的细胞毒性较低;当浓度在1-50μg/mL范围内时,四种细胞的存活率均在80%以上。普鲁士蓝染色实验和MRI成像实验表明,细胞对LP-PEI1800-SPION具有显著的摄取和成像能力。综上所述,本实验成功制备了可视化的聚阳离子脂质siRNA递送载体(LP-PEI1800-SPION),为肝癌的进一步诊疗提供了研究基础。