| 王梓霖,郭鑫宇,徐桂英,高 国,李康安.PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301探针的初步研究[J].,2023,(10):1809-1814 |
| PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301探针的初步研究 |
| The Study of PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301 Nanoprobes |
| 投稿时间：2022-12-25 修订日期：2023-02-10 |
| DOI：10.13241/j.cnki.pmb.2023.10.002 |
| 中文关键词: 分子探针 自然杀伤细胞 免疫治疗 |
| 英文关键词: Molecular probe Natural killer cell Immunotherapy |
| 基金项目:国家自然科学基金项目（81972872） |
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| 中文摘要: |
| 摘要 目的：制备PEG-G5.NH2-FITC-DOTA(Gd)- Monalizumab/IPH4301纳米探针，并研究其与NK-92MI细胞的结合能力、毒性、细胞体外MRI成像能力、促进NK-92MI细胞对MDA-MB-231三阴性乳腺癌细胞的杀伤能力。方法：制备PEG-G5.NH2-FITC-DOTA(Gd)- Monalizumab/IPH4301纳米探针，利用透射电子显微镜（Transmission Electron Microscope，TEM）进行表征，并使用ImageJ对其粒径进行统计；通过流式细胞术分析纳米材料和NK-92MI细胞的结合能力；应用MRI对纳米探针标记的NK-92MI细胞体外成像并分析其T1加权（T1WI）的信号改变。利用蛋白印迹（Western blot）检测经过与NK-92MI细胞共培养后三阴性乳腺癌MDA-MB-231细胞凋亡相关蛋白的表达以及Elisa法检测培养体系内的IFN-γ的表达量。结果：制备的纳米探针颗粒粒径分布均匀，形态近似球形。其与NK-92MI细胞结合的量以及T1信号随着纳米探针浓度增加而逐渐增加。使用Monalizumab和IPH4301两种抗体修饰的纳米探针增强了NK-92MI细胞释放IFN-γ并促进MDA-MB-231乳腺癌细胞的凋亡。结论：PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301纳米探针在结合NK-92MI细胞后，可以在3.0T磁共振下进行体外成像，并增强了NK-92MI细胞对MDA-MB-231细胞的杀伤能力。 |
| 英文摘要: |
| ABSTRACT Objective: To synthesize PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301 nanoprobes, and investigate the ability of PEG-G5.NH2-FITC-DOTA(Gd)- Monalizumab/IPH4301 nanoprobe bound to NK-92MI cells, effect on cell viability, magnetic resonance imaging in vitro and promotion of the apoptosis of targeted tumor cells by NK-92MI cells. Methods: PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/IPH4301 nanoprobes were synthesized and characterized by Transmission Electron Microscope (TEM) and its particle size was calculated using Image J software. Furthermore, flow cytometry was used to analyze the cellular uptake of nanoprobes to NK-92MI cells and magnetic resonance imaging was applied to measure the T1 signal of NK-92MI cells treated with different concentration of nanoprobes. Finally, the expressions of apoptosis-related proteins, such as Cleaved-caspase3, Bax, Bcl-2, were investigated by Western Blot in one of triple-negative breast cancer cell line (MDA-MB-231) after co-culture with NK-92MI cells at ratio of 1:5 overnight and the concentrations of interferon-γ were detected by Elisa assay in the medium. Results: The prepared nanoprobes had uniform particle size distribution and the morphology was nearly spherical. The amounts of nanoprobes bound to NK-92MI cells and T1 signal detected by MRI gradually increased with rising concentration. Moreover, nanoprobes modified with Monalizumab and IPH4301 antibodies promoted the release of interferon-γ in the medium and the apoptosis of MDA-MB-231 cells after cocultured with NK-92MI cells overnight. Conclusion: After binding to NK-92MI cells, the PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab/ IPH4301 nanoprobes could be visualized with 3.0T MRI in vitro and enhanced cytotoxic effect of NK-92MI cells to induce the apoptosis of MDA-MB-231 cells. |
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