Volume 21, Issue 1 (2018)
mjms 2018, 21(1): 35-40 |
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Keshtdar F, Ramezani R. Expression Level of miR-9 in Exosomes Derived from Ovarian Epithelial Carcinoma Cells and the Effects of Exosome Treatment on VEGF Expression in Human Umbilical Vein Endothelial Cells. mjms 2018; 21 (1) :35-40
URL: http://mjms.modares.ac.ir/article-30-22504-en.html
URL: http://mjms.modares.ac.ir/article-30-22504-en.html
1- Department of Genetics, Faculty of Advanced Science and Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran , Iran
2- Department of Biomedical Sciences, Women Research Center, Alzahra University, Tehran, Iran
2- Department of Biomedical Sciences, Women Research Center, Alzahra University, Tehran, Iran
Abstract: (8733 Views)
Aims: Exosomes are considered as a protective and enriched source of shuttle microRNAs. However, the precise biological mechanism of exosomal microRNAs in recipient cells remains to be further clarified. The aim of this study waz to investigate Expression level of miR-9 in exosomes derived from ovarian epithelial carcinoma cells and the effects of exosome treatment on Vascular Endothelial Growth Factor )VEGF( expression in human umbilical vein endothelial cells.
Material and Methods: Exosomes were purified from the conditioned medium of ovarian epithelial carcinoma cells. Exosome size and morphology were examined by a scanning electron microscope. Purified exosomes were labeled with PKH26 red fluorescent labeling kit, then labeled exosomes were incubated with human umbilical vein endothelial cells (HUVECs) for 12h at 37°C, and the cellular uptake was monitored using an inverted fluorescence microscope. Expression levels of miR-9 and VEGF were measured by real-time PCR. Paired t-test was used for data analysis.
Findings: The purified MSCs-derived exosomes had a spherical shape with a diameter between 50 to 100nm. PKH26-labeled exosomes can be taken up by SKOV3 tumor cells with high efficiency. The expression levels of miR-9 in both ovarian tumor cells and their exosomes. Exosomes derived from ovarian tumor cells caused increased expression of VEGF in exosome-treated endothelial cells.
Conclusion: Exosomes derived from ovarian tumor cells led to increased expression of VEGF in endothelial cells. As miR-9 was enriched in both ovarian tumor cells and their exosomes, it seems that exosomal transfer of miR-9 may affect the expression of VEGF in endothelial cells during tumor angiogenesis.
Material and Methods: Exosomes were purified from the conditioned medium of ovarian epithelial carcinoma cells. Exosome size and morphology were examined by a scanning electron microscope. Purified exosomes were labeled with PKH26 red fluorescent labeling kit, then labeled exosomes were incubated with human umbilical vein endothelial cells (HUVECs) for 12h at 37°C, and the cellular uptake was monitored using an inverted fluorescence microscope. Expression levels of miR-9 and VEGF were measured by real-time PCR. Paired t-test was used for data analysis.
Findings: The purified MSCs-derived exosomes had a spherical shape with a diameter between 50 to 100nm. PKH26-labeled exosomes can be taken up by SKOV3 tumor cells with high efficiency. The expression levels of miR-9 in both ovarian tumor cells and their exosomes. Exosomes derived from ovarian tumor cells caused increased expression of VEGF in exosome-treated endothelial cells.
Conclusion: Exosomes derived from ovarian tumor cells led to increased expression of VEGF in endothelial cells. As miR-9 was enriched in both ovarian tumor cells and their exosomes, it seems that exosomal transfer of miR-9 may affect the expression of VEGF in endothelial cells during tumor angiogenesis.
Article Type: Original Research |
Subject:
Sport physiology
Received: 2018/06/27 | Accepted: 2018/06/27
Received: 2018/06/27 | Accepted: 2018/06/27
References
1. Karst AM, Drapkin R. Ovarian cancer pathogenesis: A model in evolution. J Oncol. 2010;2010:932371. [Link]
2. Rojas V, Hirshfield KM, Ganesan S, Rodriguez-Rodriguez L. Molecular characterization of epithelial ovarian cancer: Implications for diagnosis and treatment. Int J Mol Sci. 2016;17(12):2113. [Link] [DOI:10.3390/ijms17122113]
3. Folkman J. Angiogenesis. Annu Rev Med. 2006;57:1-18. [Link] [DOI:10.1146/annurev.med.57.121304.131306]
4. Makrilia N, Lappa T, Xyla V, Nikolaidis I, Syrigos K. The role of angiogenesis in solid tumours: An overview. Eur J Intern Med. 2009;20(7):663-71. [Link] [DOI:10.1016/j.ejim.2009.07.009]
5. Mathivanan S, Fahner CJ, Reid GE, Simpson RJ. ExoCarta 2012: Database of exosomal proteins, RNA and lipids. Nucleic Acids Res. 2012;40(Database issue):D1241-4. [Link]
6. Ratajczak J, Miekus K, Kucia M, Zhang J, Reca R, Dvorak P, et al. Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: Evidence for horizontal transfer of mRNA and protein delivery. Leukemia. 2006;20(5):847-56. [Link]
7. Taylor DD, Gercel-Taylor C. MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol. 2008;110(1):13-21. [Link] [DOI:10.1016/j.ygyno.2008.04.033]
8. Théry C, Ostrowski M, Segura E. Membrane vesicles as conveyors of immune responses. Nat Rev Immunol. 2009;9(8):581-93. [Link] [DOI:10.1038/nri2567]
9. Vlassov AV, Magdaleno S, Setterquist R, Conrad R. Exosomes: Current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim Biophys Acta. 2012;1820(7):940-8. [Link] [DOI:10.1016/j.bbagen.2012.03.017]
10. Yang C, Robbins PD. The roles of tumor-derived exosomes in cancer pathogenesis. Clin Dev Immunol. 2011;2011:842849. [Link]
11. Motavaf M, Pakravan K, Babashah S, Malekvandfard F, Masoumi M, Sadeghizadeh M. Therapeutic application of mesenchymal stem cell-derived exosomes: A promising cell-free therapeutic strategy in regenerative medicine. Cell Mol Biol (Noisy-le-grand). 2016;62(7):74-9. [Link]
12. Safari S, Malekvandfard F, Babashah S, Alizadeh-Asl A, Sadeghizadeh M, Motavaf M. Mesenchymal stem cell-derived exosomes: A novel potential therapeutic avenue for cardiac regeneration. Cell Mol Biol (Noisy-le-grand). 2016;62(7):66-73. [Link]
13. Babashah S. MicroRNAs: Key regulators of oncogenesis. Switzerland: Springer International Publishing; 2014. [Link]
14. Behbahani GD, Ghahhari NM, Javidi MA, Molan AF, Feizi N, Babashah S. MicroRNA-Mediated post-transcriptional regulation of epithelial to mesenchymal transition in cancer. Pathol Oncol Res. 2017;23(1):1-12. [Link]
15. Javidi MA, Ahmadi AH, Bakhshinejad B, Nouraee N, Babashah S, Sadeghizadeh M. Cell-free microRNAs as cancer biomarkers: The odyssey of miRNAs through body fluids. Med Oncol. 2014;31(12):295. [Link] [DOI:10.1007/s12032-014-0295-y]
16. Bach DH, Hong JY, Park HJ, Lee SK. The role of exosomes and miRNAs in drug-resistance of cancer cells. Int J Cancer. 2017;141(2):220-30. [Link] [DOI:10.1002/ijc.30669]
17. Zhuang G, Wu X, Jiang Z, Kasman I, Yao J, Guan Y, et al. Tumour-secreted miR-9 promotes endothelial cell migration and angiogenesis by activating the JAK-STAT pathway. EMBO J. 2012;31(17):3513-23. [Link]
18. Ma L, Young J, Prabhala H, Pan E, Mestdagh P, Muth D, et al. miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol. 2010;12(3):247-56. [Link]
19. Shigehara K, Yokomuro S, Ishibashi O, Mizuguchi Y, Arima Y, Kawahigashi Y, et al. Real-time PCR-based analysis of the human bile microRNAome identifies miR-9 as a potential diagnostic biomarker for biliary tract cancer. PLoS One. 2011;6(8):e23584. [Link]
20. Song Y, Li J, Zhu Y, Dai Y, Zeng T, Liu L, et al. MicroRNA-9 promotes tumor metastasis via repressing E-cadherin in esophageal squamous cell carcinoma. Oncotarget. 2014;5(22):11669-80. [Link]
21. Hayat Nosaeid M, Mahdian R, Jamali S, Maryami F, Babashah S, Maryami F, et al. Validation and comparison of two quantitative real-time PCR assays for direct detection of DMD/BMD carriers. Clin Biochem. 2009;42(12):1291-9. [Link]
22. Katoh M. Therapeutics targeting angiogenesis: Genetics and epigenetics, extracellular miRNAs and signaling networks (Review). Int J Mol Med. 2013;32(4):763-7. [Link] [DOI:10.3892/ijmm.2013.1444]
23. Pakravan K, Babashah S, Sadeghizadeh M, Mowla SJ, Mossahebi-Mohammadi M, Ataei F, et al. MicroRNA-100 shuttled by mesenchymal stem cell-derived exosomes suppresses in vitro angiogenesis through modulating the mTOR/HIF-1α/VEGF signaling axis in breast cancer cells. Cell Oncol (Dordr). 2017;40(5):457-70. [Link]
24. Ghosh G, Subramanian IV, Adhikari N, Zhang X, Joshi HP, Basi D, et al. Hypoxia-induced microRNA-424 expression in human endothelial cells regulates HIF-α isoforms and promotes angiogenesis. J Clin Invest. 2010;120(11):4141-54. [Link]
25. Bridge G, Monteiro R, Henderson S, Emuss V, Lagos D, Georgopoulou D, et al. The microRNA-30 family targets DLL4 to modulate endothelial cell behavior during angiogenesis. Blood. 2012;120(25):5063-72. [Link]
26. Liu Y, Luo F, Wang B, Li H, Xu Y, Liu X, et al. STAT3-regulated exosomal miR-21 promotes angiogenesis and is involved in neoplastic processes of transformed human bronchial epithelial cells. Cancer Lett. 2016;370(1):125-35. [Link]
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