MiR-21 Is Required for the Epithelial–Mesenchymal Transition in MDA-MB-231 Breast Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. miR-21 Decreased Disease-Specific and Overall Survival Rates in Breast Cancer In Vivo and In Vitro
2.2. miR-21 Downregulation Decreased the Colony Size in MDA-MB-231 Cells via Increasing Survival-Related Mechanisms
2.3. miR-21 Promotes EMT
2.4. miR-21 Affects Wnt-11-Mediated Cellular Responses
2.5. Regaining miR-21 Reverses the Metastatic Phenotype of MDA-MB-231 Cells
3. Discussion
4. Materials and Methods
4.1. Overall Survival Data
4.2. Cell Culture
4.3. RNA Extraction and qRT–PCR
4.4. CRISPR/Cas9 Assay
4.5. Genomic Cleavage Assay
4.6. Extracellular Vesicle Isolation
4.7. Nanoparticle Tracking Analysis (NTA)
4.8. Hanging Drop Assay
4.9. Colony Formation
4.10. Western Blot Analysis
4.11. Immunostaining
4.12. Plasmid Transfection for Survival Assay
4.13. Wound Healing Assay
4.14. Determination of ROS Generation
4.15. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
BCa | Breast cancer |
EMT | Epithelial-mesenchymal transition |
EV | Extracellular vesicle |
miR | microRNA |
TNBC | Triple Negative Breast Cancer |
TCGA-BRCA | The Cancer Genome Atlas Breast Invasive Carcinoma |
ROS | Reactive Oxygen Species |
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gRNA | RNA Sequence |
---|---|
miR-21 gRNA1 | CTCATGGCAACACCAGTCGA |
miR-21 gRNA2 | CTCATGGCAACACCAGTCGA |
miR-21 gRNA3 | ATGTCAGACAGCCCATCGAC |
miR-21 gRNA4 | ATGTTGACTGTTGAATCTCA |
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Arisan, E.D.; Rencuzogullari, O.; Cieza-Borrella, C.; Miralles Arenas, F.; Dwek, M.; Lange, S.; Uysal-Onganer, P. MiR-21 Is Required for the Epithelial–Mesenchymal Transition in MDA-MB-231 Breast Cancer Cells. Int. J. Mol. Sci. 2021, 22, 1557. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22041557
Arisan ED, Rencuzogullari O, Cieza-Borrella C, Miralles Arenas F, Dwek M, Lange S, Uysal-Onganer P. MiR-21 Is Required for the Epithelial–Mesenchymal Transition in MDA-MB-231 Breast Cancer Cells. International Journal of Molecular Sciences. 2021; 22(4):1557. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22041557
Chicago/Turabian StyleArisan, Elif Damla, Ozge Rencuzogullari, Clara Cieza-Borrella, Francesc Miralles Arenas, Miriam Dwek, Sigrun Lange, and Pinar Uysal-Onganer. 2021. "MiR-21 Is Required for the Epithelial–Mesenchymal Transition in MDA-MB-231 Breast Cancer Cells" International Journal of Molecular Sciences 22, no. 4: 1557. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22041557