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基于负载pEGFP/p53的超顺磁性氧化铁纳米颗粒的胶质母细胞瘤靶向基因治疗

Glioblastoma Targeted Gene Therapy Based on pEGFP/p53-Loaded Superparamagnetic Iron Oxide Nanoparticles.

作者信息

Eslaminejad Touba, Nematollahi-Mahani Seyed Noureddin, Ansari Mehdi

机构信息

Kerman University of Medical Sciences Kerman, Kerman, Iran.

出版信息

Curr Gene Ther. 2017;17(1):59-69. doi: 10.2174/1566523217666170605115829.

Abstract

BACKGROUND

Blood-brain barrier (BBB) separates the neural tissue from circulating blood because of its high selectivity. This study focused on the in vitro application of magnetic nanoparticles to deliver Tp53 as a gene of interest to glioblastoma (U87) cells across a simulated BBB model that comprised KB cells.

MATERIAL AND METHOD

After magnetic and non-magnetic nanoparticles were internalized by KB cells, their location in these cells was examined by transmission electron microscopy. Transfection efficiency of DNA to U87 cells was evaluated by fluorescence microscopy, real time PCR, flowcytometry, and Western immuno-blotting. When a magnetic field was applied, a large number of magnetic nanoparticles accumulated in KB cells, appearing as black dots scattered in the cytoplasm of cells. Fluorescence microscope examination showed that transfection of the DNA to U87 target cells was highest in cells treated with magnetic nanoparticles and exposed to a magnetic field. Also it was reflected in significantly increased mRNA level while the p53 protein level was decreased.

CONCLUSION

It could be concluded that a significant increase in total apoptosis was induced in cells by magnetic nanoparticles, coupled with exposure to a magnetic force (p ≤0.01) as compared with cells that were not exposed to magnetism.

摘要

背景

血脑屏障(BBB)因其高选择性将神经组织与循环血液分隔开来。本研究聚焦于磁性纳米颗粒在体外的应用,通过包含KB细胞的模拟血脑屏障模型,将作为感兴趣基因的Tp53递送至胶质母细胞瘤(U87)细胞。

材料与方法

磁性和非磁性纳米颗粒被KB细胞内化后,通过透射电子显微镜检查它们在这些细胞中的位置。通过荧光显微镜、实时PCR、流式细胞术和Western免疫印迹评估DNA对U87细胞的转染效率。施加磁场时,大量磁性纳米颗粒在KB细胞中聚集,表现为散布在细胞质中的黑点。荧光显微镜检查显示,在用磁性纳米颗粒处理并暴露于磁场的细胞中,DNA对U87靶细胞的转染率最高。这也反映在mRNA水平显著升高而p53蛋白水平降低。

结论

可以得出结论,与未暴露于磁场的细胞相比,磁性纳米颗粒与暴露于磁力相结合可诱导细胞总凋亡显著增加(p≤0.01)。

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