核酸适配体修饰的前列腺癌靶向载体的体内外评价  

Evaluation of aptamer-modified prostate cancer targeting vector in vitro and in vivo

作　　者：郭彦琨[1] 武鑫[1] 刘玮[1] GUO YanKun WU Xin LIU Wei(Department of Clinical Pharmacy, Shanghai General Hospital Affiliated to Shanghai J iaotong University, Shanghai 200080,Chin)

机构地区：[1]上海交通大学附属第一人民医院临床药学科,上海200080

出　　处：《药学服务与研究》2017年第3期174-178,共5页Pharmaceutical Care and Research

基　　金：国家自然科学基金(81302212);上海交通大学医工交叉项目(YG2015QN14)

摘　　要：目的:构建具有前列腺癌靶向作用纳米基因载体聚酰氨胺-聚乙二醇-适配体(PAMAM-PEG-A10-3.2),考察靶向载体合成效果,携载基因能力,入细胞机制,体内外靶向等特性。方法:用核磁共振(NMR)鉴定PAMAM-PEG-A10-3.2结构;激光粒度仪测定复合物粒径和zeta电位;透射电镜(TEM)观察纳米复合物形态;竞争性抑制实验用于考察纳米复合物入细胞机制;pEGFP-N2-luc质粒(pDNA)考察纳米复合物体外靶向性;LNCaP裸鼠移植肿瘤小鼠,考察纳米复合物体内靶向性。结果:结果显示,PAMAM-PEG-A10-3.2合成成功,可以包裹NC-miRNA,形成稳定的纳米复合物。TEM见纳米复合物成球形。靶向载体主要通过受体介导内吞入胞。体外基因转染实验显示,靶向纳米复合物可以增加对前列腺癌细胞表面特异性膜抗原(PSMA)高表达细胞(LNCaP)的转染效率,LNCaP裸鼠移植肿瘤小鼠活体成像显示,靶向载体具有良好的前列腺癌靶向性。结论:PAMAM-PEG-A10-3.2对PSMA高表达的前列腺癌细胞具有良好的体内外靶向性。Objective：To construct a nano gene vector,polyamidoamine-polyethylene glycol-aptamer（PAMAM-PEGA10-3.2）for targeting prostate cancer,and evaluate the synthesis effect,carrying capacity of gene,the mechanism of cell entry,and in vitro and in vivo targeted-effect of the nano gene vector.Methods：PAMAM-PEG-A10-3.2was characterized by1 H-nuclear magnetic resonance（1 H-NMR）.The average size and zeta potential of nanocomposities were measured by Zetasizer.Transmission electron microscope（TEM）was used to observe the morphology of the nanocomposities.Competitive inhibition assay was used to investigate the mechanism of nanocomposities into cell.pEGFP-N2-luc plasmid（pDNA）and LNCaP tumor-bearing nude mouse model were used to evaluate the in vitro and in vivo targeting,respectively.Results：The results demonstrated the successful synthesis of PAMAM-PEG-A10-3.2,which could load NC-miRNA to form stable nanocomposites.TEM showed that nanocomposites were nearly in spherical shape.The targeting vector entered cell viareceptor-mediated endocytosis.Gene transfection experiments in vitro indicated that the targeted nanocomposites could increase the transfection efficiency of the prostate specific membrane antigen（PSMA）-positive prostate cancer cells（LNCaP）.Fluorescence imaging showed that targeted nanocomposites had a potential of PCa-targeting effect in LNCaP tumor-bearing nude mouse model.Conclusion：PAMAM-PEG-A10-3.2 had a good in vitro and in vivo targeting of PSMA-positive prostate cancer cells.

关 键 词：核酸适配体 NC-miRNA 聚酰氨胺 前列腺癌细胞株 LNCaP 

分 类 号：R944.9[医药卫生—药剂学]