肝癌靶向磁共振造影剂LA-PEG-SPION的制备与评价

发布时间：2018-04-20 22:18

本文选题：肝癌 + 去唾液酸糖蛋白受体　；参考：《延边大学》2017年硕士论文

【摘要】：目的:去唾液酸糖蛋白受体(ASGP-R)是肝源性细胞表面特异性表达的受体蛋白,它能够高效地识别带有半乳糖残基的配体,并通过内化作用将其摄取。肝癌细胞表面含有大量的ASGP-R,可作为特异性靶点进行靶向药物的研究。因此,本文旨在合成一种能够特异性靶向肝癌细胞表面受体ASGP-R的磁共振造影剂(LA-PEG-SPION),并探讨其肝癌靶向能力及磁共振造影作用。方法:采用共沉淀法制备亲水性的超顺磁性氧化铁纳米粒子(SPION),以缩合反应将LA-PEG-COOH与SPION结合构建LA-PEG-SPION。利用动态光散射仪、X线衍射仪、红外光谱仪以及聚焦电子显微镜对LA-PEG-SPION进行形貌粒径的表征。MTT法考察SPION与LA-PEG-SPION对HepG2(人肝癌细胞)和Hela(人宫颈癌细胞)的细胞毒性。普鲁士蓝染色、流式细胞术检测其对肝癌细胞的靶向作用,体外磁共振成像验证其靶向磁共振造影的作用。结果:LA-PEG-SPION.在电镜下呈均匀类球形,形态规则,分散度良好;激光粒度分析仪检测LA-PEG-SPION粒径约为30±4.5 nm;电位滴定仪检测其表面电势约为31 ± 1.5mV;MTT实验显示LA-PEG-SPION对HepG2细胞及Hela细胞均无毒性或低毒性,即使药物浓度达到400 μ g/mL,其对细胞仍无明显毒性;普鲁士蓝染色可见HepG2细胞对LA-PEG-SPION的摄取量明显高于SPION,细胞内可见蓝染颗粒较对照组明显增多;流式细胞术结果可见肝癌细胞的荧光标记率达到68.8%,明显高于对照组与竞争实验组;体外磁共振成像可见HepG2细胞组T2加权信号强度较对照组明显降低。结论:成功合成出LA-PEG-SPION分子探针,为其进一步体内实验及临床研究提高实验依据。
[Abstract]:Aim: sialic acid glycoprotein receptor (ASGP-R) is a receptor protein specifically expressed on the surface of hepatogenic cells. It can efficiently recognize ligands with galactose residues and ingest them through internalization. Hepatoma cells contain a large number of ASGP-Rs on the surface, which can be used as specific targets for the study of targeted drugs. Therefore, the aim of this paper is to synthesize a kind of magnetic resonance contrast agent (La-PEG-SPION), which can specifically target ASGP-R on the surface of hepatoma cells, and to investigate its ability of targeting liver cancer and the effect of magnetic resonance imaging (MRI). Methods: hydrophilic superparamagnetic ferric oxide nanoparticles were prepared by coprecipitation method. The LA-PEG-COOH was combined with SPION to form LA-PEG-SPION by condensation reaction. The morphology and particle size of LA-PEG-SPION were characterized by X-ray diffractometer, infrared spectrometer and focused electron microscope. The cytotoxicity of SPION and LA-PEG-SPION to HepG2 (human hepatocellular carcinoma cell) and Hela (human cervical cancer cell) was investigated by MTT method. Prussian blue staining and flow cytometry were used to detect its targeting effect on hepatoma cells. In vitro magnetic resonance imaging (MRI) was used to verify the effect of targeted magnetic resonance imaging (MRI). The results showed that: LA-PEG-SPION. The particle size of LA-PEG-SPION was about 30 卤4.5 nm by laser particle size analyzer, and the surface potential was about 31 卤1.5 MV by potentiometric titration assay. The results showed that LA-PEG-SPION had no toxicity or low toxicity to HepG2 cells and Hela cells. Even if the drug concentration reached 400 渭 g 路mL ~ (-1), it still had no obvious toxicity to cells. Prussian blue staining showed that the uptake of LA-PEG-SPION by HepG2 cells was significantly higher than that of SPION cells, and the blue staining granules in cells were significantly higher than those in the control group. Flow cytometry showed that the fluorescence labeling rate of hepatoma cells was 68.8, which was significantly higher than that of the control group and the competitive experimental group, and the T2-weighted signal intensity of the HepG2 cell group was significantly lower than that of the control group. Conclusion: the molecular probe of LA-PEG-SPION was successfully synthesized, which can improve the experimental basis for further in vivo and clinical studies.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7;R445.2

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