摘要
目的 构建NHE 1反义基因磁性纳米颗粒 ,探讨以氧化铁磁性纳米颗粒为载体转染NHE 1反义基因治疗肿瘤的可能性。方法 应用共沉淀法制备外包葡聚糖的氧化铁磁性纳米颗粒(dextrancoatedironoxidenanoparticles ,DCIONP)。使用透射电镜和激光粒度检测仪检测DCIONP的形态和粒径 ,采用琼脂糖凝胶电泳分析氧化铁磁性纳米颗粒结合NHE 1反义基因及抵抗DNaseⅠ消化的能力 ,采用磁场吸引的方法分析氧化铁磁性纳米颗粒携带NHE 1反义基因定向运动的能力。结果 透射电镜观察显示 :纳米颗粒形状不规则 ,颗粒内部有一氧化铁的核心。激光粒度检测仪检测证实DCIONP的平均直径在 4 7nm左右。结合实验的琼脂糖凝胶电泳显示 ,pll DCIONP在各种质量比均有良好的DNA结合能力。DNA保护实验显示从质量比 0 .5∶1开始pll DCIONP能有效保护DNA不被DNaseⅠ降解。沉淀实验证明pll DCIONP可携带质粒DNA在外加磁场作用下在液体中定向移动。结论 成功构建了NHE 1反义基因磁性纳米颗粒 ,该颗粒具有粒径小、稳定性高及磁靶向运动的特点。
Objective To Construct Na + H + exchanger 1 (NHE 1) antisense gene magnetic nanoparticies,and evaluate feasibility of using magnetic iron oxide nanoparticies as vector to transfect NHE 1 antisense gene in therapy of cancer. Methods The dextran coated iron oxide nanoparticles (DCIONP) was synthesized with deposition. The configuration and diameter of DCIONP was detected by transmission electron microscope and zetasizer. The potential of adsorbing NHE 1 antisense gene and resisting DNase I digestion of DCIONP was analyzed by agarose gel electrophoresis. The potential of orientation movement of iron oxide nanoparticles binding NHE 1 antisense gene was analyzed by attracting of magnetic field. Results Transmission electron microscope shows: nanoparticies's shape is irregular,and there is a core of ferric oxide inside nanoparticies. Under zetasizer, the mean diameter of the DCIONP is 47 nm. Agarose gel electrophoresis of binding experiment shows that pll DCIONP have the satisfactory potential to adsorb DNA.Protection experiment shows that pll DCIONP can protect DNA from DNaseI digestion effectively. Deposition experiment shows that the directional movement of pll DCIONP binding plasmid under magnetic field in liquid. Conclusions NHE 1 antisense gene magnetic nanoparticies have be constructed successfully. Small diameter, fine stability and magnetic target movement are characteristics of NHE 1 antisense gene magnetic nanoparticies.
出处
《消化外科》
CSCD
2005年第1期44-47,共4页
Journal of Digestive Surgery
基金
第三军医大学科研基金资助项目 (2 0 0 2 )