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为得到低毒、高效的聚阳离子基因载体,以甲基丙烯酸氨乙酯(AMA)和甲基丙烯酸N,N-二乙基氨乙基酯(DEAEMA)为单体,以2-溴代异丁酸乙酯(EBIB)为引发剂,通过原子转移自由基聚合(ATRP)制备了两种聚(甲基丙烯酸氨乙酯-co-甲基丙烯酸N,N-二乙基氨乙基酯)阳离子无规共聚物(P(AMA-co-DEAEMA),简称P).琼脂糖凝胶电泳实验结果表明聚合物P作为阳离子载体可以有效地络合DNA,通过粒径仪测定的复合物粒子的尺寸在400~600 nm之间.扫描电镜观察的P/DNA复合物形貌是分散均匀的球形颗粒.以25k Da PEI为阳性参照,利用MTT比色法考察了聚合物P对HEK293T细胞的毒性.结果表明,聚合物P的细胞毒性低于25 k Da PEI的细胞毒性.以25 k Da PEI和裸质粒DNA作为参照,我们进一步考察了聚合物P与DNA形成的复合物在HEK293T细胞中的转染效率.结果表明P/DNA复合物在HEK293T细胞中的转染效率远远高于裸质粒DNA的转染效率,并且接近于25 k Da PEI/DNA复合物的转染效率.
In order to obtain low toxicity and high efficiency of polycation gene carrier, aminoethyl methacrylate (AMA) and N, N-diethylaminoethyl methacrylate (DEAEMA) as monomers, 2-bromo Two kinds of poly (aminoethyl methacrylate-co-N, N-diethylaminoethyl methacrylate) were prepared by atom transfer radical polymerization (ATRP) using ethyl butyrate (EBIB) Cationic random copolymer (P (AMA-co-DEAEMA), referred to as P) .The results of agarose gel electrophoresis showed that the polymer P can act as a cation carrier to complex DNA effectively, and the particle size of the composite particle Size ranged from 400 nm to 600 nm.The morphology of P / DNA complexes observed by scanning electron microscopy was spherical particles with uniform dispersion.The toxicity of polymer P to HEK293T cells was examined by MTT colorimetric method with 25 kDa PEI as a positive control The results showed that the cytotoxicity of polymer P was lower than that of 25 kDa PEI.With the reference of 25 kDa PEI and naked plasmid DNA, we further examined the complex of polymer P and DNA in HEK293T cells Transfection efficiency The results showed that the transfection efficiency of P / DNA complex in HEK293T cells was much higher than that of naked plasmid DNA , And close to the transfection efficiency of the 25 kDa PEI / DNA complex.