靶向纳米造影剂C225-USPIO对EGFR高表达鼻咽癌CNE1细胞系磁共振免疫显像研究

发布时间：2018-05-13 07:31

本文选题：C225 + USPIO　；参考：《华中科技大学》2010年硕士论文

【摘要】：目的制备针对表皮生长因子受体(EGFR)的靶向纳米磁共振造影剂并进行体外检测。方法将西妥昔单抗(C225)与超小型超顺磁性氧化铁(USPIO)通过碳二亚胺法连接成针对EGFR的靶向纳米造影剂C225-USPIO。选择EGFR高表达鼻咽癌CNE1细胞系,流式细胞术、间接免疫荧光检测C225-USPIO与CNE1细胞的免疫结合活性。普鲁士蓝染色以及细胞透射电镜(TEM)检测CNE1细胞对C225-USPIO靶向结合摄取情况。4.7T磁共振成像(MRI)检测Fe浓度为50μg/ml的C225-USPIO孵育CNE1细胞不同时间梯度后引起的MRI信号强度变化。结果流式细胞术、间接免疫荧光结果显示C225-USPIO对鼻咽癌CNE1细胞表面EGFR有良好的免疫结合活性。普鲁士蓝染色及细胞透射电镜(TEM)检测显示C225-USPIO可在CNE1细胞表面特异性结合EGFR并被摄取入胞内。4.7T MRI结果显示,CNE1细胞可以结合摄取C225-USPIO,引起MRI T2信号强度明显降低。结论C225-USPIO能特异性的与高表达EGFR的鼻咽癌CNE1细胞结合,具有良好的特异性和靶向性,并且可以明显的降低磁共振的T2信号强度,为以后的体内实验提供了充足的实验依据。
[Abstract]:Objective to prepare an EGF receptor EGFR targeted nano magnetic resonance contrast medium and detect it in vitro. Methods C225-USPIO-targeted nano-contrast agent C225-USPIO-targeted EGFR was prepared by ligation of cetuximab C225) and ultrasmall superparamagnetic iron oxide (USPIO) by carbodiimide method. The high expression of EGFR in nasopharyngeal carcinoma (NPC) CNE1 cell line, flow cytometry and indirect immunofluorescence assay were used to detect the immunobinding activity of C225-USPIO and CNE1 cells. Prussian blue staining and transmission electron microscopy (TEM) were used to detect the uptake of C225-USPIO targeting by CNE1 cells. 4.7T MRI) was used to detect the changes of MRI signal intensity induced by C225-USPIO incubated with 50 渭 g/ml Fe for different time gradient. Results flow cytometry and indirect immunofluorescence showed that C225-USPIO had good immunobinding activity to EGFR on the surface of nasopharyngeal carcinoma (NPC) CNE1 cells. Prussian blue staining and transmission electron microscopy (TEM) analysis showed that C225-USPIO could specifically bind to EGFR on the surface of CNE1 cells and be ingested into the cells. The results showed that C225-USPIO could be combined with the uptake of C225-USPIO. the signal intensity of MRI T2 was significantly decreased. Conclusion C225-USPIO can specifically bind to CNE1 cells of NPC with high expression of EGFR. It has good specificity and targeting, and can obviously reduce the intensity of T2 signal of MRI, which provides sufficient experimental basis for future in vivo experiments.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63

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