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目的:构建叶酸(FA)分子靶向载基质金属蛋白酶9(MMP-9)反义寡核苷酸(ASODN)磁性纳米复合物(FA-MNP-MMP-9-ASODN),研究该复合物对叶酸受体(FR)阳性鼻咽癌的靶向能力及基因转染效率。方法:采用课题组前期制备的FA靶向磁性纳米载体,通过醛基与氨基缩合反应与MMP-9-ASODN偶联,制备FA-MNP-MMP-9-ASODN,采用琼脂糖凝胶电泳分析寡核苷酸与载体的偶联。通过分析荷HNE-1及CNE-2瘤裸鼠的MRI,和铁染色、透射电镜结果研究该2种细胞及瘤块吞噬纳米复合物情况以及观察细胞内FITC分析该载体的基因转染。结果:电泳结果显示ASODN已成功连接在FA靶向磁性纳米载体上。HNE-1细胞能有效摄取该纳米复合物,细胞内见较多FITC,而CNE-2细胞不能摄取该纳米复合物,细胞内未见FITC。HNE-1瘤块也能有效地吞噬该纳米复合物。结论:成功构建FA-MNP-MMP-9-ASODN纳米复合物,具有良好的FA分子靶向性及基因转染效率。
OBJECTIVE: To construct FA-MNP-MMP-9-ASODN (MMP-9) antisense oligodeoxynucleotide targeting matrix metalloproteinase 9 (MMP-9) Targeting ability and gene transfection efficiency of folic acid receptor (FR) positive nasopharyngeal carcinoma. Methods: FA-MNP-MMP-9-ASODN was prepared by FA-targeting magnetic nanocarrier prepared in the previous study and coupled with MMP-9-ASODN by aldehyde group and amino condensation reaction. Coupling of nucleotides to a carrier. By analyzing the MRI images of HNE-1 and CNE-2 tumor-bearing nude mice and the results of iron staining and transmission electron microscopy, we observed the phagocytosis of these two kinds of cells and tumor nodules, and observed the intracellular FITC analysis of gene transfection. Results: Electrophoresis showed that ASODN was successfully linked to FA-targeting magnetic nanocarriers. HNE-1 cells effectively uptake the nanocomposites, see more FITC in cells, CNE-2 cells can not ingest the nanocomposites, no FITC in the cells. The HNE-1 tumor mass can also effectively engulf the nanocomposite. CONCLUSION: FA-MNP-MMP-9-ASODN nanocomposites have been successfully constructed and have good FA molecular targeting and gene transfection efficiency.