邓雯,吴维敏,亓立峰,赵厚育
DENG Wen,WU Weimin,QI Lifeng,ZHAO Houyu

• 1:贵州医科大学
• 2:同济大学医学院附属同济医院妇产科
• 3:同济大学医学院纳米院
• 4:贵州医科大学附院耳鼻喉科

摘要(Abstract)：

目的:构建聚乙烯亚胺(PEI)修饰的四氧化三铁(Fe_3O_4)磁性纳米粒,观察其表征,探讨其作为基因载体的可行性。方法:用PEI-Fe_3O_4磁性纳米粒包被质粒转染鼻咽癌CNEII细胞,转染miRNA-HIF-1α干扰质粒和miRNA-NC对照质粒分别为H组、NC组,空白对照组设为C组;用电子显微镜观察H组和NC组纳米粒的大小及表征,用荧光显微镜观察H组和NC组转染效率,用Real Time RT-PCR及Western Blot方法检测低氧诱导因子-1α(HIF-1α)mRNA及蛋白表达的抑制情况。结果:成功构建PEI-Fe_3O_4磁性纳米粒;H组和NC组的转染率分别为(63.6±4.2)%、(65.8±6.1)%,差异无统计学意义(P>0.05);H组HIF-1αmRNA及蛋白表达抑制率与NC组、C组比较,差异具有统计学意义(P<0.05)。结论:PEI-Fe_3O_4磁性纳米粒是一种较为理想的基因转染载体。
Objective: To construct PEI-Fe_3O_4 nanoparticles and observe its characteristics,and to investigate the feasibility of PEI-Fe_3O_4 nanoparticles as carrier in transfection. Methods: Plasmids coated with PEI-Fe_3O_4 magnetic nanoparticels were transfected into CNEII cells,cells transfected with miRNA-HIF-1α plasmid as group H while cells transfected with miRNA-NC plasmid as NC group,and CNEII cells without transfection served as C group. Observing size and morphology of PEI-Fe_3O_4 nanoparticles with scanning electronic microscope. Using fluorescence microscope to measure efficiency of transfection. Proteins were detected by western blot and mRNAs were detected by RT-PCR. Results:PEI-Fe_3O_4 was successfully constructed. The transfection efficiency of group H and group NC was( 63. 6 ± 4. 2) % and( 65. 8 ± 6. 1) %,difference was not statistically significant( P > 0. 05). The difference of suppression ratio of protein and mRNA of HIF-1α was significant compared to group C and group NC( P < 0. 05). Conclusion: PEI-Fe_3O_4 magnetic nanoparticles can be used as an ideal carrier of gene transfection.

关键词(KeyWords)： 乙烯亚胺-四氧化三铁;磁性纳米粒;基因转染;鼻咽癌;缺氧诱导因子-1α;信使RNA
PEI-Fe3O4 magnetic nanoparticles;gene transfection;nasopharyngeal carcinoma;hypoxia inducible factor-1α;message RNA

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区科学基金项目(NO.81260353)

作者(Author): 邓雯,吴维敏,亓立峰,赵厚育
DENG Wen,WU Weimin,QI Lifeng,ZHAO Houyu

DOI: 10.19367/j.cnki.1000-2707.2016.04.004

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