双嵌段磷酸胆碱基聚合物作为c-myc AS-ODN基因运输载体研究

目的 探讨双嵌段磷酸胆碱基聚合物MPC30-DEA70 作为反义寡核苷酸（AS-ODN）运输载体的安全性和有效性.方法 将载体MPC30-DEA70（N）与c-myc AS-ODN（P）在不同N/P比值的条件下复合,应用DNA凝胶电泳法、荧光显微镜、流式细胞术和四甲基偶氮唑盐（MTT）法对复合物溶液进行检测,观察复合物在HeLa细胞中的转染情况.结果 c-myc AS-ODN能与载体MPC30-DEA70电性中和,N/P比值越大复合物正电性越强;MPC30-DEA70/AS-ODN复合物能够进入细胞,转染效率随着N/P比值增大而增大;同时随着N/P比值增大复合物对细胞的增殖抑制作用显著增强.结论 MPC30-DEA70可以有效负载和运输c-myc AS-ODN,抑制HeLa细胞增殖,是一种新型的、安全有效的非病毒类转基因载体.

Study of phosphorylcholine diblock copolymer as c-myc antisense oligodeoxynucleotide gene carrier

Objective To investigate the safety and efficacy of a novel cationic phosphorylcholine diblock copolymer as antisense oligodeoxynucleotide gene carrier. Methods MPC30 - DEA70/c - myc AS - ODN complexes were compounded at different N/P ratios （cationic/anionic）. The solution of the complexes were determined using DNA gel electrophoresis, fluorescent microscopy, FCM and MTT assay. The transfection of the complexes in HeLa cells was observed. Results c - mye AS - ODN could neutralize charge of the carrier MPC30 - DEA70, with a significant positive correlation between N/P ratios and the enhanced positive charge of the complexes ; The complexes could be transfered into the Hela cells and the transfection efficiency were significantly enhanced with the increase of N/P ratios as well. Conclusion This novel cationic phosphorylcholine copolymer can effectively load and transport human c -myc antisense oligodoxynu- cleotides, which may act as an efficient and safe non - viral gene transfer vector.