Jiaxing University, Jiaxing, Zhejiang, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(10):5703-5713. doi: 10.26355/eurrev_202005_21362.
To study the effectiveness of natural killer cell-derived exosome (NK-Exos)-entrapped paclitaxel (PTX-NK-Exos) in enhancing its anti-tumor effect.
The NK-Exos were isolated through ultra-high-speed centrifugation, and the PTX-NK-Exos system was constructed via electroporation. The morphology, particle size, Zeta potential and entrapment rate of PTX-NK-Exos were evaluated using transmission electron microscope (TEM), dynamic light scattering (DLS), Western blotting and high-performance liquid chromatography (HPLC), respectively. The uptake of Exos in human breast cancer MCF-7 cells was observed under a laser confocal microscope. Moreover, the effect of PTX-NK-Exos on MCF-7 cell viability was determined through methyl thiazolyl tetrazolium (MTT) assay, flow cytometry and 4',6-diamidino-2-phenylindole (DAPI) staining. The effects of PTX-NK-Exos on messenger ribonucleic acid (mRNA) and protein expressions of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and Caspase-3 in MCF-7 cells were detected using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting, respectively.
The NK-Exos were successfully isolated via ultra-high-speed centrifugation, and they had uniform particle size and high expression of markers for Exos. MCF-7 cells could take up Exos. The PTX-NK-Exos drug delivery system was successfully prepared using electroporation. In PTX group and NK-Exos group, the proliferation of MCF-7 cells declined, the nuclear apoptosis was evident and the apoptosis rate of MCF-7 cells rose compared with those in Control group. In PTX group and PTX-NK-Exos group, the migration of MCF-7 cells declined compared with that in Control group. According to the results of qRT-PCR and Western blotting, PTX-NK-Exos exerted an anti-tumor effect through inducing the up-regulation of Bax and Caspase-3 in the apoptotic signaling pathway in tumor cells.
Exos isolated through ultra-high-speed centrifugation can be used to prepare the PTX-NK-Exos drug delivery system through electroporation. Drug-loaded Exos can effectively inhibit proliferation and induce apoptosis of tumor cells, thereby exerting an anti-tumor effect.
研究自然杀伤细胞衍生的外泌体（NK-Exos）包载紫杉醇（PTX-NK-Exos）增强其抗肿瘤作用的效果。
通过超速离心分离 NK-Exos，通过电穿孔构建 PTX-NK-Exos 系统。使用透射电子显微镜（TEM）、动态光散射（DLS）、Western 印迹和高效液相色谱（HPLC）分别评估 PTX-NK-Exos 的形态、粒径、Zeta 电位和包封率。使用激光共聚焦显微镜观察 Exos 在人乳腺癌 MCF-7 细胞中的摄取情况。此外，通过噻唑蓝（MTT）测定法、流式细胞术和 4',6-二脒基-2-苯基吲哚（DAPI）染色来确定 PTX-NK-Exos 对 MCF-7 细胞活力的影响。使用定量逆转录聚合酶链反应（qRT-PCR）和 Western 印迹分别检测 PTX-NK-Exos 对 MCF-7 细胞中 B 细胞淋巴瘤-2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）和 Caspase-3 的信使核糖核酸（mRNA）和蛋白表达的影响。
通过超速离心成功分离 NK-Exos，其具有均匀的粒径和 Exos 的高表达标志物。MCF-7 细胞可以摄取 Exos。使用电穿孔成功制备了 PTX-NK-Exos 药物递送系统。与对照组相比，在 PTX 组和 NK-Exos 组中，MCF-7 细胞的增殖减少，核凋亡明显，MCF-7 细胞的凋亡率升高。与对照组相比，在 PTX 组和 PTX-NK-Exos 组中，MCF-7 细胞的迁移减少。根据 qRT-PCR 和 Western 印迹的结果，PTX-NK-Exos 通过诱导肿瘤细胞凋亡信号通路中 Bax 和 Caspase-3 的上调发挥抗肿瘤作用。
通过超速离心分离的 Exos 可通过电穿孔用于制备 PTX-NK-Exos 药物递送系统。载药 Exos 可有效抑制肿瘤细胞增殖并诱导其凋亡，从而发挥抗肿瘤作用。
