外泌体作为一种将生物制剂和小分子抗癌药物共运输的纳米载体，目前尚处于襁褓期。因此，在本研究中，我们探究了来自人类脐血干细胞的外泌体作为生物制造纳米载体，来共同运输肿瘤抑制子miR-125a和微管不稳定剂紫杉醇（Docetaxel，DTX），以瞄准小鼠三阴性乳腺癌4T1细胞的增殖和迁移侵袭性。在本研究中，我们成功将miR-125a转染到hUCBMSCs中。然后，在经过优化的温和超声处理孵育技术的作用下，将DTX封装到非转染和转染外泌体中。通过MTT和形态测定法比较hUCBMSC Exo-DTX和miR-125a Exo-DTX的抗癌效果。通过体外划痕愈合和Transwell侵袭实验验证了后者卓越的抗转移行为。此外，通过共聚焦显微镜和FESEM实验进一步证实了miR-125a和DTX的协同作用。 hUCBMSC外泌体表现出8.86±1.97ng DTX/μg外泌体和miRNA保存容量相等于12.31±5.73%的DTX载荷。联合载药方案（miR-125a Exo-DTX）在4T-1细胞中表现出IC50为192.8ng/ml，几乎比游离DTX IC50（472.8ng/mL）低2.36倍。此外，miR-125a Exo-DTX治疗导致伤口扩大至6.14±0.38%，而DTX和miR-125a外泌体单独治疗分别导致伤口闭合率为18.71±4.5%和77.36±10.4%。miR-125a Exo-DTX治疗进一步表现为4T1细胞侵袭性显著降低（降低了3.5±1.8%），以及细胞骨架降解和核变形。miR-125a表达的DTX载药外泌体清楚地展示了miR-125a Exo-DTX的协同凋亡和抗迁移效率，这种协同的抗癌和抗转移作用归因于hUCBMSC来源外泌体的DTX和miR-125a作为载荷。 Copyright © 2023 Elsevier Inc. 发布。

Exosomes, as the nanocarrier for the co-delivery of biologicals and small anticancer molecules is yet at its infancy. Herein, we investigated hUCBMSC derived exosomes as a biogenic nanocarrier for the co-delivery of tumor suppressor miR-125a and microtubule destabilizing Docetaxel (DTX) to target the proliferative and migratory aggressiveness of the murine TNBC 4T1 cells.In this study, hUCBMSCs from the human umbilical cord blood cells (hUCB) were successfully transfected with miR-125a. Thereafter, DTX was encapsulated into both non-transfected and transfected exosomes by optimized mild sonication-incubation technique. The anticancer efficiency of hUCBMSC Exo-DTX and miR-125a Exo-DTX were compared by MTT and morphometric assay. The prominent anti-metastatic behaviour of the latter was confirmed by in-vitro wound healing and transwell invasion assay. Further, the synergistic effect of miR-125a and DTX was confirmed by F-actin and nuclear degradation by confocal and FESEM assay.hUCBMSC exosomes exhibited DTX payload of 8.86 ± 1.97 ng DTX/ μg exosomes and miRNA retention capacity equivalent to 12.31 ± 5.73 %. The co-loaded formulation (miR-125a Exo-DTX) exhibited IC50 at 192.8 ng/ml in 4T-1 cells, which is almost 2.36 folds' lower than the free DTX IC50 (472.8 ng/mL). Additionally, miR-125a Exo-DTX treatment caused wound broadening upto 6.14±0.38 % while treatment with free DTX and miR-125a exosomes alone caused 18.71±4.5 % and 77.36±10.4 % of wound closure respectively in 36 h. miR-125a Exo-DTX treatment further exhibited significantly reduced invasiveness of 4T1 cells (by 3.5 ± 1.8 %) along with prominent cytoskeletal degradation and nuclear deformation as compared to the miR-125a exosomes treated group. The miR-125a expressing DTX loaded exosomal formulation clearly demonstrated the synergistic apoptotic and anti-migratory efficiency of the miR-125a Exo-DTX.The synergistic anticancer and anti-metastatic effect of miR-125a Exo-DTX resulted in due to presence of both DTX and miR-125a as the cargo of hUCBMSC derived exosomes.Copyright © 2023. Published by Elsevier Inc.