Delivery of miR-424-5p via Extracellular Vesicles Promotes the Apoptosis of MDA-MB-231 TNBC Cells in the Tumor Microenvironment
Abstract
:1. Introduction
2. Results
2.1. PD-L1 Expression Is Increased in TNBC
2.2. PD-L1 Is Directly Regulated by miR-424-5p
2.3. EVs Are Secreted into AT-MSC Culture Supernatant
2.4. EVs-Mediated Delivery of miR-424 Decreases PD-L1 Expression in Recipient Cells
2.5. miR-424-5p Regulates Cytokine Secretion
2.6. miR-424-5p Upregulation Promotes Cytotoxicity against Tumors
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Culture
4.2. Preparation of EVs
4.3. Electron Microscopy
4.4. Measurement of the Size Distribution and Particle Number by NTA
4.5. Real-Time Reverse Transcription-PCR
4.6. Cell Transfection
4.7. Dual-Luciferase Reporter Assay
4.8. AGO2-RNA Immunoprecipitation
4.9. Western Blotting
4.10. Tumor Cell/T Cell Coculture Model
4.11. Immunofluorescence
4.12. Apoptosis Assessment of Tumor Cells in the Coculture System
4.13. Caspase 3/7 Assay
4.14. LDH Release Assay
4.15. In Vivo Studies
4.16. Statistics
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Zhou, Y.; Yamamoto, Y.; Takeshita, F.; Yamamoto, T.; Xiao, Z.; Ochiya, T. Delivery of miR-424-5p via Extracellular Vesicles Promotes the Apoptosis of MDA-MB-231 TNBC Cells in the Tumor Microenvironment. Int. J. Mol. Sci. 2021, 22, 844. https://doi.org/10.3390/ijms22020844
Zhou Y, Yamamoto Y, Takeshita F, Yamamoto T, Xiao Z, Ochiya T. Delivery of miR-424-5p via Extracellular Vesicles Promotes the Apoptosis of MDA-MB-231 TNBC Cells in the Tumor Microenvironment. International Journal of Molecular Sciences. 2021; 22(2):844. https://doi.org/10.3390/ijms22020844
Chicago/Turabian StyleZhou, Yueyuan, Yusuke Yamamoto, Fumitaka Takeshita, Tomofumi Yamamoto, Zhongdang Xiao, and Takahiro Ochiya. 2021. "Delivery of miR-424-5p via Extracellular Vesicles Promotes the Apoptosis of MDA-MB-231 TNBC Cells in the Tumor Microenvironment" International Journal of Molecular Sciences 22, no. 2: 844. https://doi.org/10.3390/ijms22020844