Synthesis of cell penetrating peptide decorated magnetic nanoparticles loading cisplatin for nasopharyngeal cancer therapy
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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72 h后（载体铁浓度10 μg/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。Abstract: 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 FT-IR. 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 on 293T cells 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.
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Key words:
- magnetic nanoparticles /
- drug delivery system /
- cell penetrating peptide /
- TAT
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