Exosome Production, Isolation and Characterization from A549 Epithelial Carcinoma Cells

Yıl 2019, Cilt: 47 Sayı: 4, 383 - 388, 01.12.2019

Esra Mutlu Özge Kaya Arzu Birinci Yildirim Ayhan Çetinkaya

https://doi.org/10.15671/hjbc.520101

Cited By: 3

Öz

Exosomes
are natural nanoparticles that special features have been started to be used
for recent years in the treatment of cancer as a natural, homogeneous,
nanosized, targeted vesicles (~ 50- 100 nm). They have high avidity (many
conformational attachment) to attach onto targeted cancer cell surfaces. They
are composed of bioactive double-layered lipid layers in which their original
nature has the adhesive proteins interacting with the cancer cell membrane
easily. In this study, the exosomes of non-small cell lung cancer A549
epithelial carcinoma cells were investigated for their potential to be the
natural or synthetic drug carrier. Firstly, exosomes of A549 cell line were
produced in exosome-free media. Immediately after, isolation of their exosomes
were realized by using ultracentrifugation procedure. SEM image, particle size,
zeta potential, exosomal RNA analysis and Protein Content by Bradford assays
were performed. Findings (Size: 168 nm; zeta: -16mV) on the properties of A549
cell exosomes proved their potential to be used as the drug carrier for cancer
cell therapy.

Anahtar Kelimeler

Exosome, ultracentrifugation, A549 Cell Line, miRNA

Kaynakça

• 1. G. Mignot, S. Roux, C. Thery, E. Ségura, L. Zitvogel, Prospects for exosomes in immunotherapy of cancer, J. Cell. Mol. Med., 10 (2006) 376-388.
• 2. M. Morello, V. Minciacchi, P. De Candia, J. Yang, E. Posadas, H. Kim, D. Griffiths, N. Bhowmick, L. Chung, P. Gandellini, Large oncosomes mediate intercellular transfer of functional microRNA, Cell cycle, 12 (2013) 3526-3536.
• 3. P. Vader, X. O. Breakefield, M. J. Wood, Extracellular vesicles: emerging targets for cancer therapy, Trends. Mol. Med., 20 (2014) 385-393.
• 4. M. S. Kim, M. J. Haney, Y. Zhao, V. Mahajan, I. Deygen, N. L. Klyachko, E. Inskoe, A. Piroyan, M. Sokolsky, O. Okolie, Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells, Nanomed-Nanotechnol., 12 (2016) 655-664.
• 5. A. Jemal, R. Siegel, E. Ward, Y. Hao, J. Xu, T. Murray, M. J. Thun, Cancer statistics, 2008, CA: a cancer journal for clinicians, 58 (2008) 71-96.
• 6. R. Siegel, C. DeSantis, A. Jemal, Colorectal cancer statistics, 2014, CA: a cancer journal for clinicians, 64 (2014) 104-117.
• 7. C. Théry, M. Ostrowski, E. Segura, Membrane vesicles as conveyors of immune responses, Nat. Rev. Immunol., 9 (2009) 581.
• 8. J. Palma, S. C. Yaddanapudi, L. Pigati, M. A. Havens, S. Jeong, G. A. Weiner, K. M. E. Weimer, B. Stern, M. L. Hastings, D. M. Duelli, MicroRNAs are exported from malignant cells in customized particles, Nucleic Acids Res., 40 (2012) 9125-9138.
• 9. S. El-Andaloussi, Y. Lee, S. Lakhal-Littleton, J. Li, Y. Seow, C. Gardiner, L. Alvarez-Erviti, I. L. Sargent, M. J. Wood, Exosome-mediated delivery of siRNA in vitro and in vivo, Nat. Protoc., 7 (2012) 2112.
• 10. C. Lässer, M. Eldh, J. Lötvall, Isolation and characterization of RNA-containing exosomes, Jove-J. Vis. Exp., (2012).
• 11. M. M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 72 (1976) 248-254.
• 12. W. Li, D. Mu, F. Tian, Y. Hu, T. Jiang, Y. Han, J. Chen, G. Han, X. Li, Exosomes derived from Rab27a‑overexpressing tumor cells elicit efficient induction of antitumor immunity, Mol. Med. Rep., 8 (2013) 1876-1882.
• 13. M. Wysoczynski,M. Z. Ratajczak, Lung cancer secreted microvesicles: underappreciated modulators of microenvironment in expanding tumors, Int. J. Cancer, 125 (2009) 1595-1603.
• 14. M. A. Morse, J. Garst, T. Osada, S. Khan, A. Hobeika, T. M. Clay, N. Valente, R. Shreeniwas, M. A. Sutton, A. Delcayre, A phase I study of dexosome immunotherapy in patients with advanced non-small cell lung cancer, J. Transl. Med., 3 (2005) 9.
• 15. Y.-T. Tang, Y.-Y. Huang, L. Zheng, S.-H. Qin, X.-P. Xu, T.-X. An, Y. Xu, Y.-S. Wu, X.-M. Hu, B.-H. Ping, Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum, Int. J. Mol. Med., 40 (2017) 834-844.