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中国生物医学工程学报  2024, Vol. 43 Issue (6): 712-719    DOI: 10.3969/j.issn.0258-8021.2024.06.008
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核酸适配体介导的氧化铁纳米颗粒对4T1乳腺癌细胞的热疗效应
陈恩远1,2&, 蒋政廷3&, 丁佳怡1, 阚俊楠1, 祖涵瑜1, 闫鹏3*
1(滨州医学院医学影像学院,山东 烟台 264003)
2(苏州大学附属第一医院,江苏 苏州 215000)
3(滨州医学院基础医学院,山东 烟台 264003)
Hyperthermia Effect of Fe3O4 Nanoparticles Guided by Aptamer on 4T1 Breast Cancer Cells
Chen Enyuan1,2&, Jiang Zhengting3&, Ding Jiayi1, Kan Junnan1, Zu Hanyu1, Yan Peng3*
1(School of Medical Imaging, Binzhou Medical University, Yantai 264000, Shandong, China)
2(First Affiliated Hospital of Soochow University, Suzhou 215000, Jiangsu, China)
3(School of Basic Medical Sciences, Binzhou Medical University, Yantai 264003, Shandong, China)
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摘要 磁性纳米材料因其独特的磁学性质在肿瘤影像诊断、磁热疗等领域获得了广泛应用。本研究采用化学共沉淀法制备了Fe3O4纳米颗粒,并借助酰胺反应将核酸适配体(Apt23)偶联在颗粒表面,形成了钙网蛋白受体靶向的氧化铁纳米颗粒(Fe3O4@Apt),并进一步研究了Fe3O4@Apt纳米颗粒对4T1乳腺癌细胞的热疗效应。通过X射线衍射分析Fe3O4纳米颗粒的物相和晶体结构;采用透射电子显微镜、纳米粒度仪和振动样品磁强计对修饰前后的Fe3O4纳米颗粒的形貌、粒径、zeta电位及饱和磁化强度进行理化性质表征;通过傅里叶变换红外光谱对修饰前后Fe3O4纳米颗粒的有效官能团进行检测;通过普鲁士蓝染色测定修饰先后的Fe3O4纳米颗粒对4T1细胞的靶向作用;采用MTT法检测4T1细胞和小鼠胚胎成纤维细胞(MEF)在修饰前后Fe3O4纳米颗粒上的存活与增殖;在交变磁场(ACMF)的作用下,测定修饰前后的Fe3O4纳米颗粒的升温性能及体外磁热疗效果;所有定量检测实验中至少设3个重复样。结果显示,Fe3O4纳米颗粒的结晶性良好,修饰前后Fe3O4纳米颗粒的形貌和磁学性能未发生明显变化。核酸适配体修饰增大了颗粒的水合粒径,使表面负电性增强。Fe3O4和Fe3O4@Apt的平均粒径分别为(9±4)和(18±5)nm。Zeta电位分别为(-20.4±0.6)和(-27±0.4)mV。Fe3O4@Apt纳米颗粒在1 037 cm-1处的吸收峰也说明Apt23成功偶联到Fe3O4纳米颗粒。修饰前后的Fe3O4纳米颗粒可支持4T1和MEF细胞的正常生长和增殖,在培养24 h后,不同浓度的Fe3O4@Apt纳米颗粒组与Fe3O4纳米颗粒组相比无显著性差异(P>0.05)。在ACMF的作用下,修饰前后的Fe3O4纳米颗粒升温效果良好,且靶向治疗组的细胞增殖率低于对照组(P<0.05)。Fe3O4@Apt对乳腺癌细胞4T1具有良好的靶向效应,且磁热疗在体外能有效抑制4T1细胞增殖。
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陈恩远
蒋政廷
丁佳怡
阚俊楠
祖涵瑜
闫鹏
关键词 Fe3O4纳米颗粒Apt23适配体磁热疗乳腺癌    
Abstract:Magnetic nanomaterials have been widely used in tumor imaging diagnosis and magnetic hyperthermia due to their unique magnetic properties. In this study, Fe3O4 nanoparticles were prepared by chemical coprecipitation method, and calreticulin receptor-targeted iron oxide nanoparticles (Fe3O4@Apt) were formed by coupling aptamer (Apt23) on the surface of the nanoparticles through an amide reaction. The hyperthermia effects of Fe3O4@Apt nanoparticles on breast cancer tumor cells (4T1) were investigated. The phase and crystal structure of Fe3O4 nanoparticles were analyzed by X-ray diffraction. The morphology, particle size, zeta potential and saturation magnetization of Fe3O4 nanoparticles before and after modification were characterized by transmission electron microscope, nanoparticle size analyzer and vibrating sample magnetometer, respectively. Functional groups on Fe3O4 nanoparticles, both before and post-modification, were analyzed through Fourier transform infrared spectroscopy. The targeting effect of Fe3O4 nanoparticles on 4T1 cells was determined by Prussian blue staining. Cell viability and proliferation of 4T1 cells and mouse embryonic fibroblasts (MEF) were detected by MTT assay. Under an alternating magnetic field (ACMF), the heating performance and in vitro magnetic hyperthermia effects of Fe3O4 nanoparticles before and after modification were measured. There were at least 3 replicates (n≥3) in all quantitative tests. Experimental results showed that the crystallization of Fe3O4 nanoparticles was good, and the morphology and magnetic properties of Fe3O4 nanoparticles did not change significantly before and after modification. The aptamer modification enhanced the surface electronegativity and increased the average particle size. The average particle sizes of Fe3O4 and Fe3O4@Apt were (9±4) and (18±5) nm, respectively. The zeta potentials were (-20.4±0.6) and (-27±0.4) mV, respectively. The absorption peak of Fe3O4@Apt nanoparticles at1 037 cm-1 indicated successful coupling with Apt23. Fe3O4 nanoparticles before and after modification could support the normal growth and proliferation of 4T1 and MEF cells. After 24 h incubation, there was no significant difference of the cells viability between Fe3O4@Apt nanoparticles and Fe3O4 nanoparticles at different concentrations (P>0.05). Under the ACMF, Fe3O4 nanoparticles exhibited effective heating capabilities before and after modification, and the cell proliferation rate in the targeted treatment group was lower than that in the control group (P<0.05). In conclusion, Fe3O4@Apt nanoparticles exhibited excellent targeting specificity toward breast cancer cell 4T1, and the magnetic hyperthermia effectively inhibited 4T1 cell proliferation in vitro.
Key wordsFe3O4 nanoparticles    Apt23 aptamer    magnetic hyperthermia    breast cancer
收稿日期: 2023-02-05     
PACS:  R318  
基金资助:国家自然科学基金(51401031); 山东省大学生创新创业训练计划项目(S202110440011)
通讯作者: *E-mail: phyyp@bzmc.edu.cn   
作者简介: &共同第一作者
引用本文:   
陈恩远, 蒋政廷,丁佳怡, 阚俊楠, 祖涵瑜, 闫鹏. 核酸适配体介导的氧化铁纳米颗粒对4T1乳腺癌细胞的热疗效应[J]. 中国生物医学工程学报, 2024, 43(6): 712-719.
Chen Enyuan, Jiang Zhengting, Ding Jiayi, Kan Junnan, Zu Hanyu, Yan Peng. Hyperthermia Effect of Fe3O4 Nanoparticles Guided by Aptamer on 4T1 Breast Cancer Cells. Chinese Journal of Biomedical Engineering, 2024, 43(6): 712-719.
链接本文:  
http://cjbme.csbme.org/CN/10.3969/j.issn.0258-8021.2024.06.008     或     http://cjbme.csbme.org/CN/Y2024/V43/I6/712
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