Quan Stephen, Kumar Piyush, Narain Ravin
Donadeo Innovation Centre for Engineering, Department of Chemical and Materials Engineering, and ‡Department of Oncology, Faculty of Medicine & Dentistry, University of Alberta, Edmonton, Alberta, Canada.
Donadeo Innovation Centre for Engineering, Department of Chemical and Materials Engineering, and Department of Oncology, Faculty of Medicine & Dentistry, University of Alberta, Edmonton, Alberta, Canada.
ACS Biomater Sci Eng. 2016 May 9;2(5):853-859. doi: 10.1021/acsbiomaterials.6b00085. Epub 2016 Apr 25.
Glycopolymers of statistical and block configurations were synthesized from 2-lactobionamidoethyl methacrylamide (LAEMA) and 2-aminoethyl methacrylamide hydrochloride (AEMA) by the reversible addition-fragmentation chain transfer (RAFT) polymerization. These cationic glycopolymers were found to form very stable polyplexes with EGFR siRNA as determined by dynamic light scattering and agarose gel electrophoresis. The polyplexes revealed to be very stable even in the presence of serum proteins. Transfection studies of the glycopolymer-EGFR siRNA polyplexes were achieved in HeLa cells to determine the EGFR knockdown efficiency, cellular uptake and cytotoxicity. In this study, the block copolymer with the shortest AEMA segment was the most effective in EGFR gene silencing, however this block copolymer revealed to be slightly more toxic as compared to the statistical copolymers studied at higher w/w ratios. In addition, gene silencing of up to 80-85% was achieved with this low-molecular-weight block copolymer.
通过可逆加成-断裂链转移（RAFT）聚合反应，由甲基丙烯酸2-乳糖酰胺基乙酯（LAEMA）和甲基丙烯酸2-氨基乙酯盐酸盐（AEMA）合成了无规和嵌段构型的糖聚合物。通过动态光散射和琼脂糖凝胶电泳测定，发现这些阳离子糖聚合物与表皮生长因子受体（EGFR）小干扰RNA（siRNA）形成非常稳定的多聚体。即使在存在血清蛋白的情况下，这些多聚体也显示出非常稳定。在HeLa细胞中进行了糖聚合物-EGFR siRNA多聚体的转染研究，以确定EGFR基因敲低效率、细胞摄取和细胞毒性。在本研究中，具有最短AEMA链段的嵌段共聚物在EGFR基因沉默方面最有效，然而，与在较高重量比下研究的无规共聚物相比，这种嵌段共聚物显示出稍高的毒性。此外，使用这种低分子量嵌段共聚物实现了高达80-85%的基因沉默。
