Leptin Modulates Exosome Biogenesis in Breast Cancer Cells: An Additional Mechanism in Cell-to-Cell Communication
Abstract
:1. Introduction
2. Experimental Section
2.1. Antibodies and Reagents
2.2. Cell Cultures
2.3. Isolation of Tumor-Derived Exosomes
2.4. Transmission Electron Microscopy (TEM)
2.5. Nanoparticle Tracking Analysis (NTA)
2.6. Acetylcholinesterase Assay (AchE) to Measure Exosomes
2.7. TUNEL Assay
2.8. Immunoprecipitation and Immunoblot Analysis
2.9. Total RNA Extraction and Reverse Transcription PCR Assay
2.10. Proximity Ligation Assay
2.11. Statistical Analyses
3. Results
3.1. Leptin Induces Generation of Exosomes in Breast Cancer Cells
3.2. Leptin Upregulates Tsg101 Expression in Breast Cancer Cells
3.3. Tsg101 Protein Directly Interacts with Chaperon Protein Hsp90 in Breast Cancer Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Giordano, C.; Gelsomino, L.; Barone, I.; Panza, S.; Augimeri, G.; Bonofiglio, D.; Rovito, D.; Naimo, G.D.; Leggio, A.; Catalano, S.; et al. Leptin Modulates Exosome Biogenesis in Breast Cancer Cells: An Additional Mechanism in Cell-to-Cell Communication. J. Clin. Med. 2019, 8, 1027. https://doi.org/10.3390/jcm8071027
Giordano C, Gelsomino L, Barone I, Panza S, Augimeri G, Bonofiglio D, Rovito D, Naimo GD, Leggio A, Catalano S, et al. Leptin Modulates Exosome Biogenesis in Breast Cancer Cells: An Additional Mechanism in Cell-to-Cell Communication. Journal of Clinical Medicine. 2019; 8(7):1027. https://doi.org/10.3390/jcm8071027
Chicago/Turabian StyleGiordano, Cinzia, Luca Gelsomino, Ines Barone, Salvatore Panza, Giuseppina Augimeri, Daniela Bonofiglio, Daniela Rovito, Giuseppina Daniela Naimo, Antonella Leggio, Stefania Catalano, and et al. 2019. "Leptin Modulates Exosome Biogenesis in Breast Cancer Cells: An Additional Mechanism in Cell-to-Cell Communication" Journal of Clinical Medicine 8, no. 7: 1027. https://doi.org/10.3390/jcm8071027