Yoon Chang S, Jung Hye S, Kim Tae K, Kwon Min J, Kim Mi K, Lee Minhyung, Koh Kyung S, Rhee Byung D, Park Jeong H
Molecular Therapy Laboratory, Department of Internal Medicine, College of Medicine, Paik Institute for Clinical Research, Inje University, Busan, South Korea.
J Drug Target. 2008 Dec;16(10):773-9. doi: 10.1080/10611860802470549.
The objective of this study is to evaluate transfection efficiency and safety for gene delivery by sonoporation in comparison with cationic polymer gene carrier branched polyethylenimine (BPEI).
The cDNA expressing VEGF(165) was cloned under chicken beta-actin promoter. The plasmid DNA was transfected into the CHO, HEK293, and NIH3T3 cells using microbubble-based sonoporation and BPEI (25 kDa) under various conditions. Enzyme-linked immunosorbent assay (ELISA) was used to determine the expressed protein level. Cytotoxicities of transfection methods were compared by Cell Counting Kit-8.
At 1 MHz intensity, transfection efficiency of sonoporation was enhanced by microbubble concentration with no detrimental effects. By contrast, BPEI exacerbated cell viability, despite its high transgene expression efficiency.
Sonoporation gene therapy might be the safest technique to be used in actual clinical practice.
本研究的目的是与阳离子聚合物基因载体支链聚乙烯亚胺（BPEI）相比，评估通过声孔效应进行基因递送的转染效率和安全性。
在鸡β-肌动蛋白启动子的控制下克隆表达VEGF(165)的cDNA。在各种条件下，使用基于微泡的声孔效应和BPEI（25 kDa）将质粒DNA转染到CHO、HEK293和NIH3T3细胞中。采用酶联免疫吸附测定（ELISA）来测定表达的蛋白水平。通过细胞计数试剂盒-8比较转染方法的细胞毒性。
在1 MHz强度下，声孔效应的转染效率随微泡浓度的增加而提高，且无有害影响。相比之下，BPEI尽管具有较高的转基因表达效率，但会加剧细胞活力的下降。
声孔效应基因治疗可能是实际临床实践中最安全的技术。
