基于苯丙氨酸的聚酯酰胺纳米递药系统制备及其体外抗前列腺癌的研究

目的探究基于苯丙氨酸的聚酯酰胺的纳米递药系统的制备及其对体外培养前列腺癌细胞的抑制效果。 方法采用纳米沉淀法制备聚酯酰胺纳米粒,利用透射电镜和纳米电位仪测量纳米粒的形貌和大小分布,利用破碎沉淀法与荧光磷光光谱仪测定其载药量和包封率。采用四甲基偶氮唑盐微量酶反应比色法测定载药纳米粒对前列腺癌细胞LNCaP的抑制效果,采用膜联蛋白V-APC/7AAD双染法并用流式细胞仪测定其对肿瘤细胞的凋亡诱导作用。 结果采用纳米沉淀法成功制备了空白及包载多柔吡星（DOX）的载药聚酯酰胺纳米粒,粒径约为90～110 nm,大小分布较为均匀。空白纳米粒在不同浓度下均显示出良好的生物相容性,高浓度100 μg/ml的材料作用下,LNCaP的细胞活力仍保持在（95.32±3.97）%。与DOX组的IC50 （3.29±0.63）μg/ml相比,载药纳米粒对前列腺癌细胞活性抑制明显,IC50为（1.21±0.43）μg/ml,差异具有统计学意义（t=6.693,P<0.001）。流式细胞仪检测载药纳米粒能有效诱导前列腺癌细胞进入中晚期凋亡,对照组、DOX组与DOX@8p6组凋亡率分别为2.32%、29.16%和61.62%,后两组间差异具有统计学意义（χ²=2217,P<0.001）,且DOX@8p6组凋亡情况多于DOX组（t=11.238,P<0.001）,显示出载药纳米粒对前列腺癌细胞LNCaP更高的促凋亡效率。 结论基于苯丙氨酸的聚酯酰胺纳米粒具有良好的生物相容性,其作为递送抗肿瘤药物多柔吡星的纳米药物载体时能有效地直接抑制前列腺癌肿瘤细胞活性及诱导肿瘤细胞凋亡。

Construction and in vitro antitumor effect on prostate cancer of phenylalanine-based poly(ester amide)snanoparticles drug delivery system

ObjectiveTo study the construction of phenylalanine-based poly(ester amide)sas drug delivery system and its inhibition of in vitro prostate cancer cell LNCaP. MethodsThe phenylalanine-based poly(ester amide)s nanoparticles were synthesized by nanoprecipitation. The morphology and size distribution of nanoparticle were inspected by transmission electron microscopy and nanoparticle size analyzer. Drug loading and encapsulation rate were measured using fragmentation precipitation method by fluorescent phosphorescence spectrometer. Inhibition of prostate cancer cell LNCaP using doxorubicin-loaded phenylalanine-based poly(ester amide)s nanoparticle were inspected by MTT method and cell apoptosis effect was analyzed by V-APC/7AAD double staining method. ResultsThe phenylalanine-based poly(ester amide)s nanoparticles were synthesized successfully as doxorubicin delivery system with a uniform size of about 90-110 nm. The blank nanoparticles showed a great biocompatibility at different concentrations. The cell viability of LNCaP remained at (95.32±3.97)% at a high blank nanoparticle concentration of 100 μg/ml. Compared with the DOX group whose IC50 was (3.29±0.63) μg/ml, the activity of prostate cancer cells co-cultured with drug-loaded nanoparticles was significantly inhibited, whose IC50 was (1.21±0.43) μg/ml, which was statistically different (t= 6.693, P<0.001). The cell apoptosis effectively induced by drug-loaded nanoparticles was measured by flow cytometry detection. The control group, DOX group and DOX@8p6 group were 2.32%, 29.16% and 61.62%, respectively, while the difference between the three groups was statistically significant (F=294.44, P<0.001), and the DOX@8p6 group had more apoptosis than the DOX group (t=11.238, P<0.001), showing drug-loaded nanoparticles had higher pro-apoptotic efficiency against prostate cancer cell line LNCaP. ConclusionIt demonstrated that the biocompatible phenylalanine-based poly(ester amide)s nanoparticles as doxorubicin delivery system could provide effective prostate cancer cell inhibition and high cell apoptosis.