穿膜肽修饰的载顺铂磁性纳米复合物的制备及其对鼻咽癌的体外效应  

Synthesis of cell penetrating peptide decorated magnetic nanoparticles loading cisplatin for nasopharyngeal cancer therapy

作　　者：黎权明 钟颖 翁欢欢 苗湘琬 张娟[1] 谢慧芬 谢民强[1,2] LI Quanming;ZHONG Ying;WENG Huanhuan;MIAO Xiangwan;ZHANG Juan;XIE Hui fen;XIE Minqiang(Department of Otorhinolaryngology Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, 510000, China;Department of Otorhinolaryngology Head and Neck Surgery, Zhuhai People＇s Hospital)

机构地区：[1]南方医科大学珠江医院耳鼻咽喉头颈外科,广州510000 [2]珠海市人民医院耳鼻咽喉头颈外科

出　　处：《临床耳鼻咽喉头颈外科杂志》2018年第13期963-968,共6页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金(No:81673031;No:81372477);广东省科技计划项目(No:2017A010103010)

摘　　要：目的:制备具有穿膜作用的载顺铂磁性纳米复合物,并探究其对鼻咽癌的体外作用。方法:以醛基化海藻酸钠(ASA)改性的磁性纳米粒(ASA-MNPs)作为药物载体,将端氨基聚乙二醇(PEG)化的穿膜肽(TAT)与ASA通过醛胺缩合链接,组装成TAT修饰的载顺铂磁性纳米复合物,并依据配位络合的原理偶联顺铂(TAT-ASA-MNP@CDDP)。通过核磁氢谱、红外光谱等对载药磁性纳米复合物进行表征,通过荧光标记观察其穿膜能力,评估其生物相容性,采用CCK-8细胞毒性实验及流式细胞术评估其对鼻咽癌CNE-2细胞的抑制作用。结果:核磁氢谱和红外光谱显示TAT-ASA-MNP@CDDP分别具有TAT、PEG、ASA特征峰,其水流动力学粒径为(145.9±1.5)nm,zeta电位为(-21.66±1.24)mV,顺铂载量为(25.03±3.05)%。荧光标记显示,CNE-2能快速摄取TAT-ASA-MNP@CDDP。TAT-ASA-MNPs载体细胞毒性实验显示,共培养72h后(载体铁浓度10μg/ml),293T细胞的细胞存活率大于70%,人红细胞凝聚试验阴性。体外细胞毒性实验及凋亡实验显示,在顺铂浓度相对较低时,TAT-ASA-MNP@CDDP对CNE-2细胞的抑制作用比ASA-MNP@CDDP大(P<0.05)。结论:单纯载体无明显细胞毒副作用,生物相容性良好,成功制备的TAT-ASA-MNP@CDDP对CNE-2细胞具有明显体外抑制效应。Objective：To synthesize cisplatin loaded and cell penetrating peptide TAT decorated magnetic nanoparticles and to observe the inhibiting effect in vitro on nasopharyngeal cancer therapy.Method：The aldehyde sodium alginate coated magnetic nanoparticles（ASA-MNPs）was prepared as the drug delivery system,which was covalently attached by PEGylation TAT（TAT-ASA-MNPs）via condensation of aldehyde with amino group and then coordinated with cisplatin（TAT-ASA-MNPs@CDDP）.The complex was characterized by H NMR and FTIR.The cell penetrating ability and biocompatibility were observed by means of fluorescent tags.The inhibited effect on nasopharyngeal cancer CNE-2 cells was measured by cellular toxicity research and flow cytometry.Result：The H NMR and FT-IR of TAT-ASA-MNPs exhibited the characteristic peaks of TAT,PEG as well as ASA.The dynamic light scattering showed the hydrodynamic diameter of the complex was（145.9±1.5）nm.Zeta potential was（-21.66±1.24）mV and the drug loading rate was（25.03±3.05）%.Fluorescent labeling assay revealed that FITC marked TAT-ASA-MNPs was quickly taken up by CNE-2 cells.Cytotoxicity experiment on293 Tcells displayed high survival rate（〉70%）after cultured for 72 h.Negative hemagglutination reflected decent biocompatibility.In vitro cytotoxicity-test and cell apoptosis assay exhibited obvious inhibition on CNE-2 cell with TAT-ASA-MNPs@CDDP at low concentration of cisplatin compared to ASA-MNPs@CDDP（P〈0.05）.Conclusion：TAT-ASA-MNPs showed decent biocompatibility while distinctly inhibit CNE-2 cells in vitro study.

关 键 词：磁性纳米材料 药物载体 穿膜肽TAT 鼻咽癌 

分 类 号：R739.6[医药卫生—肿瘤]