IR780-VPN靶向纳米粒的制备及其对肾母细胞瘤的抑制作用

目的制备同时携带IR780碘化物和硫酸长春新碱（VCR）的聚乳酸-羟基乙酸（PLGA）靶向纳米粒（IR780-VPN），并观察其在体外对肾母细胞瘤（SK-NEP1）的靶向作用及抗肾母细胞瘤增殖效率。 方法制备IR780-VPN，检测其基本理化特性，培养SK-NEP1细胞作为体外肿瘤细胞模型，研究IR780-VPN的体外靶向性，观察以下6组的体外抗SK-NEP1细胞增殖效率：A组，PBS（对照组）；B组，游离VCR；C组，空白纳米粒（PN）；D组，载VCR纳米粒（VPN）；E组，载IR780纳米粒（IR780-PN）；F组，IR780-VPN。 结果制备出的IR780-VPN平均粒径为（290.82±3.22）nm，粒径的分散指数（PDI）为0.024，平均Zeta电位为（1.16±0.87）mV。用细胞膜绿色荧光探针染色后于倒置荧光显微镜下观察，IR780-VPN大小均匀、形态规则，无聚集、粘连；透射电镜下观察IR780-VPN为球形或类球形，表面光滑，粒径分布均匀。其平均包封率为72.80%，平均载药量为2.80%，平均连靶率为91.30%。体外寻靶实验中可见SK-NEP1细胞表面有大量的IR780-VPN聚集。体外抗SK-NEP1细胞增殖实验证实，在相同药物质量浓度条件下，IR780-VPN组较VCR、VPN组的体外抗肿瘤细胞增殖效率高（P均<0.001），且游离VCR、VPN、IR780-VPN对SK-NEP1细胞增殖的抑制率具有浓度依赖性，药物质量浓度越大，对细胞的抑制作用越强（F=13254.105、54354.510、1370.059，P均<0.001）；而随着药物质量浓度增加，PN、IR780-PN组的细胞增殖抑制率差异均无统计学意义（P均>0.05），PBS（对照组）对SK-NEP1细胞增殖的抑制率为（2.83±0.32）%。 结论本实验成功制备了性质稳定的靶向IR780-VPN，对肾母细胞瘤细胞有良好的的靶向性，并提高了体外抗肾母细胞瘤增殖效率，为体内的肿瘤分子靶向研究提供了实验基础。

Preparation of IR780 and vincristine-loaded poly (lactic-co-glycolic acid) nanoparticles and antinep-hroblastoma cells proliferation in vitro

ObjectiveTo prepare IR780 and vincristine (VCR)-loaded poly(lactic-co-glycolic acid)(PLGA) nanoparticles (IR780-VPN) and verify their specific targeting ability and anti-proliferative efficiency in vitro. MethodsIR780-VPN were prepared to detect their basic physical and chemical characteristics. SK-NEP1 cells were used as a tumor cell model to verify the specific targeting ability and the anti-proliferative efficiency of IR780-VPN. SK-NEP1 cells were divided into six groups: group A: PBS; group B: free VCR; group C: PLGA nanoparticles; group D: VCR-PLGA nanoparticles; group E: IR780-PLGA nanoparticles; group F: IR780-VPN. ResultsThe average particle size of IR780-VPN was (290.82±3.22) nm, the particle dispersion index (PDI) was 0.024, and the average zeta potential of IR780-VPN was (1.16±0.87) mV. IR780-VPN showed stable physical and chemical properties, and they were spherical, uniform in size, and regular in shape, without aggregation oradhesion under a microscope. The morphology of IR780-VPN was spherical or quasi-spherical with a smooth surface, and the particle size distribution was uniform. The average encapsulation efficiency of IR780-VCR-PLGA nanoparticles was about 72.80%, the average loading efficiency was 2.80%, and the specific targeting efficiency was 91.30%. Many IR780-VPN were observed to target tumor cells in vitro. The cytotoxicity was the highest in group F when compared with group B and group D at the same concentration (P<0.05), and cytotoxicity was concentration-dependent in group B, group D, and group F: the higher the drug concentration, the stronger the cytotoxicity (F=13254.105, 54354.510, and 1370.059, P<0.05). There was no significant difference in group C and group E with the increase of drug concentration (P>0.05). The rate of reduced cell proliferation in group A was (2.83±0.32)%. ConclusionIR780-VPN with stable properties have been successfully prepared, which can specifically target tumor cells and have improved anti-proliferative efficiency in vitro.