CAJ | 학술논문

目的探讨偶联叶酸(FA)的载质粒PEG-PEI(-FA)-Alg-氧化铁磁性纳米复合物对叶酸受体(FR)阳性鼻咽癌(NPC)细胞的分子靶向能力及基因转染效率.方法体外实验采用铁染色和透射电镜方法观察FR阳性NPC细胞HNE-1及FR阴性NPC细胞CNE-2吞噬磁性纳米复合物的情况,并通过观察两种细胞内质粒绿色荧光蛋白(GFP)表达及流式细胞仪分析该磁性纳米复合物的基因转染效率；体内实验采用MRI、铁染色及透射电镜观察尾静脉注射磁性纳米复合物后,同时荷HNE-1及CNE-2两种瘤的同一裸鼠种植瘤细胞吞噬磁性纳米复合物的情况.结果体外实验显示,HNE-1细胞能有效摄取磁性纳米复合物,细胞内见较强GFP表达,基因转染效率为40.0％±3.0％.而CNE-2细胞不能有效摄取磁性纳米复合物,细胞内仅见很少量的GFP表达,基因转染效率为4.5％±1.5％.体内实验显示,HNE-1种植瘤细胞能有效的吞噬磁性纳米复合物,而CNE-2种植瘤内吞噬的磁性纳米复合物很少.结论偶联FA的载质粒PEG-PEI (-FA)-Alg-氧化铁磁性纳米复合物具有良好的分子靶向性及较高的基因转染效率.
목적 탐토우련협산(FA)적재질립PEG-PEI(-FA)-Alg-양화철자성납미복합물대협산수체(FR)양성비인암(NPC)세포적분자파향능력급기인전염효솔.방법 체외실험채용철염색화투사전경방법관찰FR양성NPC세포HNE-1급FR음성NPC세포CNE-2탄서자성납미복합물적정황,병통과관찰량충세포내질립록색형광단백(GFP)표체급류식세포의분석해자성납미복합물적기인전염효솔；체내실험채용MRI、철염색급투사전경관찰미정맥주사자성납미복합물후,동시하HNE-1급CNE-2량충류적동일라서충식류세포탄서자성납미복합물적정황.결과 체외실험현시,HNE-1세포능유효섭취자성납미복합물,세포내견교강GFP표체,기인전염효솔위40.0％±3.0％.이CNE-2세포불능유효섭취자성납미복합물,세포내부견흔소량적GFP표체,기인전염효솔위4.5％±1.5％.체내실험현시,HNE-1충식류세포능유효적탄서자성납미복합물,이CNE-2충식류내탄서적자성납미복합물흔소.결론 우련FA적재질립PEG-PEI (-FA)-Alg-양화철자성납미복합물구유량호적분자파향성급교고적기인전염효솔.
Objective To evaluate the molecular targeting capacity and gene transfection efficiency of magnetic nanocomposite coupling with folic acid (FA) of PEG-PEI(-FA)-Alg-Fe3O4 adsorbing plasmid to folate receptor (FR) positive nasopharyngeal carcinoma (NPC) cell.Methods The phagocytosis of magnetic nanocomposite was examined in FR positive NPC HNE-1 and FR negative NPC CNE-2 cells.Prussian blue staining,transmission electron microscope (TEM) and flow cytometry were employed to analyze its gene transfection efficiency by observing the plasmid expression of green fluorescent protein (GFP) in two cell types.Nude mice with implanted HNE-1 and CNE-2 tumor received a tail vein injection of magnetic nanocomposite.And the phagocytosis of magnetic nanocomposite was observed by magnetic resonance imaging (MRI),iron staining and TEM.Results HNE-1 cell could effectively ingest magnetic nanocomposite,there was a strong intracellular expression of GFP and the gene transfection efficiency was approximately 40.0％ ± 3.0％.However,CNE-2 cell had virtually no uptake of magnetic nanocomposite,there was a weak intracellular expression of GFP and the gene transfection efficiency was approximately 4.5％ ± 1.5％.The implanted HNE-1 tumor cell could efficiently swallow magnetic nanocomposite,but there was little phagocytosis of magnetic nanocomposite in implanted CNE-2 tumor cell.Conclusion The magnetic nanocomposite coupling with FA of PEG-PEI (-FA)-Alg-Fe3Oa adsorbing plasmid has excellent molecular targeting capacity and higher gene transfection efficiency.