Doxorubicin-loaded NK exosomes enable cytotoxicity against triple-negative breast cancer spheroids

Document Type : Original Article

Authors

1 ATMP Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran

2 Pathology Department, Dalhousie University, Halifax, Canada

3 Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran

4 Biomaterials and Tissue Engineering Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran, Iran

5 Department of Biology Sciences, Shahid Rajaee Teacher Training University, Tehran, Iran

6 Cancer and Cell Therapy Research Center, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): Natural killer (NK) cells are the most professional innate immune cells that initiate extracellular apoptosis via cytotoxic granules in malignant cells. Antitumoral properties of NK-derived exosomes (Exos) are attributed to their parent cells. Loading drugs into Exos as a carrier can enhance their effect and enable targeted delivery. In the present study, we aim to deliver Doxorubicin (DOX) to the breast cancer spheroids by NK-Exos.
Materials and Methods: Peripheral blood mononuclear cells (PBMC) were used to harvest NK cells, and NK-Exos were isolated from NK cell expansion medium using an Exo-spinTM kit. DOX was loaded via the ultrasonication method. AO/EtBr, Annexin/PI, DAPI, MTT, and spheroids of human breast cancer were used to track the cytotoxic effect of DOX-NK-Exos. The colony formation assay, scratch and transwell assays, Real-Time PCR for p53 and VEGF-A, and WB for protein expression were also performed.
Results: When compared to free DOX, all viability tests validated the inhibitory effects of DOX-NK-Exos. The obtained results indicated that DOX-NK-Exos selectively reduced tumor cell viability and spared fibroblast and MCF-10A as noncancerous cells. Long after spheroid treatment, DOX-NK-Exos’ remarkable effect persisted.
Conclusion: Human breast carcinoma mass treated with DOX-NK-Exos underwent apoptosis and showed a strong inhibitory effect on proliferation. Thus, they can reduce the side effects of chemotherapeutics and can be used as drug carriers with selective toxicity. Additionally, the additive action of this combination formula results in a more severe loss in cell viability.

Keywords

Main Subjects

References

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Iranian Journal of Basic Medical Sciences
Volume 27, Issue 12
December 2024
Pages 1604-1615

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