Author:
Useckaite Zivile,Mukhopadhya Anindya,Moran Barry,O’Driscoll Lorraine
Abstract
Abstract
MET pathway is an important actionable target across many solid tumour types and several MET inhibitors have been developed. Extracellular vesicles (EVs) are proposed to be mini-maps of their cells of origin. However, the potential of EVs to report on the MET status of their cells of origin is unknown. After applying three proposed methods of EV separation from medium conditioned by three cell lines of known MET status, this study used an extensive range of methodologies to fundamentally characterise the resulting particles (nanoparticle tracking analysis, TEM, flow cytometry, immunoblotting) and their MET status (RT-qPCR and ELISAs). The results indicated that ultracentrifugation on density-gradient (UC-DG) consistently produced the most reliable data with regards to purest EVs. EV cargo reflected MET mRNA, total MET and pMET status of their cells of origin. In conclusion, to simply determine if the general contents of conditioned medium reflect the MET status of the conditioning cells, choice of method for initial EV separation may not be crucial. However, to be confident of specifically studying EVs and thus EV-MET cargo, UC-DG followed by extensive EV characterisation is necessary.
Funder
Irish Research Council
Science Foundation Ireland
Publisher
Springer Science and Business Media LLC
Reference27 articles.
1. Lutterbach, B. et al. Lung cancer cell lines harboring MET gene amplification are dependent on Met for growth and survival. Cancer Res. 67(5), 2081–2088 (2007).
2. Zhao, Y. et al. Met tyrosine kinase inhibitor, PF-2341066, suppresses growth and invasion of nasopharyngeal carcinoma. Drug Des. Dev. Ther. 9, 4897–4907 (2015).
3. Hartmann, S., Bhola, N. E. & Grandis, J. R. HGF/met signaling in head and neck cancer: Impact on the tumor microenvironment. Clin. Cancer Res. 22(16), 4005–4013 (2016).
4. Giannelli, G. et al. Role of epithelial to mesenchymal transition in hepatocellular carcinoma. J. Hepatol. 65(4), 798–808 (2016).
5. Refaat, T. et al. c-Met overexpression in cervical cancer, a prognostic factor and a potential molecular therapeutic target. Am. J. Clin. Oncol. 40(6), 590–597 (2017).
Cited by
4 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献