WIN55,212-2-Induced Expression of Mir-29b1 Favours the Suppression of Osteosarcoma Cell Migration in a SPARC-Independent Manner
Abstract
:1. Introduction
2. Results
2.1. WIN Treatment Reduces the Migratory Ability of MG63 Cells and Affects MMP Activity
2.2. WIN Treatment Prevents SPARC Release in the Extracellular Environment and Increases the Release of Extracellular Vesicles
2.3. Silencing SPARC did not Affect the Anti-Migratory Effect of WIN
2.4. WIN Induces the Upregulation of miR-29b1 which Plays a Role in Cell Migration
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Cultures
4.3. Wound Healing Assay
4.4. Gelatin Zymography
4.5. Western Blot Analysis
4.6. Isolation and Quantification of Extracellular Vesicles from Culture Media
4.7. Gene Silencing Using siRNA
4.8. Real-Time PCR for miR-29b1 Expression
4.9. Stable Transfection of MG63 Cells with miR-29b1 Plasmid Vector
4.10. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
SPARC | Secreted protein acidic and rich in cysteine |
MMPs | Matrix metalloproteinases |
EVs | Extracellular vesicles |
AchEase | Acetylcholinesterase |
THG | Thapsigargin |
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Notaro, A.; Emanuele, S.; Geraci, F.; D’Anneo, A.; Lauricella, M.; Calvaruso, G.; Giuliano, M. WIN55,212-2-Induced Expression of Mir-29b1 Favours the Suppression of Osteosarcoma Cell Migration in a SPARC-Independent Manner. Int. J. Mol. Sci. 2019, 20, 5235. https://doi.org/10.3390/ijms20205235
Notaro A, Emanuele S, Geraci F, D’Anneo A, Lauricella M, Calvaruso G, Giuliano M. WIN55,212-2-Induced Expression of Mir-29b1 Favours the Suppression of Osteosarcoma Cell Migration in a SPARC-Independent Manner. International Journal of Molecular Sciences. 2019; 20(20):5235. https://doi.org/10.3390/ijms20205235
Chicago/Turabian StyleNotaro, Antonietta, Sonia Emanuele, Fabiana Geraci, Antonella D’Anneo, Marianna Lauricella, Giuseppe Calvaruso, and Michela Giuliano. 2019. "WIN55,212-2-Induced Expression of Mir-29b1 Favours the Suppression of Osteosarcoma Cell Migration in a SPARC-Independent Manner" International Journal of Molecular Sciences 20, no. 20: 5235. https://doi.org/10.3390/ijms20205235
APA StyleNotaro, A., Emanuele, S., Geraci, F., D’Anneo, A., Lauricella, M., Calvaruso, G., & Giuliano, M. (2019). WIN55,212-2-Induced Expression of Mir-29b1 Favours the Suppression of Osteosarcoma Cell Migration in a SPARC-Independent Manner. International Journal of Molecular Sciences, 20(20), 5235. https://doi.org/10.3390/ijms20205235