靶向血管内皮生长因子受体2超声造影剂的制备及其体外寻靶能力实验研究

发布时间：2018-04-23 10:00

本文选题：靶向超声造影剂 + 生物素-亲和素　；参考：《肿瘤影像学》2016年04期

【摘要】：目的:制备靶向血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)超声造影剂,评价其物理性质及对肿瘤细胞的寻靶能力。方法:利用生物素-亲和素桥接法将微米级超声造影剂USphere与VEGFR2抗体结合,制备靶向VEGFR2超声造影剂,检测其粒径分布;用免疫荧光法检测不同细胞株(高转移肝肿瘤细胞MHCC-97H及正常肝细胞LO2)中VEGFR2表达强度,以及靶向与非靶向VEGFR2超声造影剂对MHCC-97H和LO2细胞的结合能力。结果:靶向VEGFR2超声造影剂微泡分布均匀,平均粒径(1 012.67±78.59)nm,免疫荧光法显示VEGFR2抗体可特异性连接于微泡表面。标记荧光的VEGFR2抗体在MHCC-97H细胞高表达,在LO2细胞低表达。免疫荧光定量显示靶向VEGFR2造影剂在MHCC-97H细胞中的荧光强度比值(微泡荧光强度/细胞荧光强度)为0.75±0.32,显著高于其他3组(P0.01)。结论:本研究成功制备靶向VEGFR2超声造影剂,其大小稳定均一,可与高表达VEGFR2的MHCC-97H细胞特异性结合。
[Abstract]:Aim: to prepare 2(vascular endothelial growth factor receptor 2 VEGFR2 ultrasound contrast agent and evaluate its physical properties and targeting ability to tumor cells. Methods: the micron ultrasound contrast agent (USphere) was combined with VEGFR2 antibody by biotin-avidin bridging method to prepare the targeted VEGFR2 ultrasound contrast agent and its particle size distribution was detected. The expression of VEGFR2 in different cell lines (high metastatic liver tumor cells MHCC-97H and normal hepatocytes LO2) and the binding ability of targeted and non-targeted VEGFR2 ultrasound contrast agents to MHCC-97H and LO2 cells were detected by immunofluorescence assay. Results: the average diameter of VEGFR2 ultrasound contrast agent was 1 012.67 卤78.59 nm. Immunofluorescence assay showed that VEGFR2 antibody could be specifically linked to the surface of microbubble. The fluorescent VEGFR2 antibody was highly expressed in MHCC-97H cells and low in LO2 cells. Immunofluorescence quantitative analysis showed that the fluorescence intensity ratio of targeted VEGFR2 contrast agent in MHCC-97H cells was 0.75 卤0.32, which was significantly higher than that in other three groups (P 0.01). Conclusion: in this study, the target VEGFR2 ultrasound contrast agent was successfully prepared, and its size was stable and uniform, which could bind to MHCC-97H cells with high expression of VEGFR2.
【作者单位】：复旦大学附属中山医院超声科;江苏省南京鼓楼医院超声科;
【基金】：国家自然科学基金资助项目(No:81371577,81571676)
【分类号】：R943;R96

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