Abstract
Purpose
Oral cancer (OC) patients are at high risk to develop recurrent disease or secondary primary cancers with no available biomarkers to detect these events until a visible lesion is readily present and diagnosed by biopsy. Exosomes secreted by cancer cells are involved in tumor growth, invasion and metastasis. We aimed to determine morphological and molecular differences between oral fluid (OF)-derived exosomes of OC patients and those isolated from healthy individuals (HI).
Methods
OF from OC patients (n = 36) and HI (n = 25) was initially assessed by nanoparticle tracking analysis (NTA). Following ultracentrifugation, exosomal pellets of OC patients and HI were morphologically examined by transmission electron microscopy and atomic force microscopy (AFM). Enzyme-linked immunosorbent assay (ELISA) and western blotting (WB) were used to analyze the expression of exosomal markers—CD9, CD81 and CD63.
Results
NTA showed that OC samples of OF had a significantly higher concentration of nanoparticles/ml (p = 0.01) and modal nanoparticle size (p = 0.002) compared to HI. The difference in size was structurally highlighted by AFM three-dimensional images applied on exosomal pellets. ELISA and WB showed differential expression of exosomal markers in OC exosomes compared to HI: lower expression of CD81 and CD9 in contrast to a higher expression of CD63 (~53 kDa).
Conclusions
OF-derived exosomes from OC patients differ both morphologically and molecularly from exosomes present in HI. This study is a baseline that provides a starting point for finding exosomal biomarkers for early detection of malignant changes in high-risk patients without overt clinical signs/lesions.
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Acknowledgments
The authors would like to thank Mrs. Ronit Galron from the Department of Neurobiology, Faculty of Life Sciences, Tel Aviv University for her valuable technical assistance, Dr. Artium Khatchtouriants, from The Center for Nanoscience and Nanotechnology, Tel Aviv University, for his AFM analysis and Mr. Ariel Roytman, from Merkel Technologies, Yehud, Israel, for his assistance with the NTA. Special thanks go to Dr. Ran Yahalom, Head of the Department of Oral and Maxillofacial Surgery, Sheba Medical Center, for his assistance in collecting the saliva samples. The authors also want to thank Ms. Esther Eshkol for editorial assistance. The study was supported in part by the Dave and Sarah Babish Fund in Oral Pathology, Tel- Aviv University.
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. For this type of study, formal consent is not required.
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This work was performed in partial fulfillment of the requirements for a Ph.D. degree of Ayelet Zlotogorski- Hurvitz, Sackler Faculty of Medicine, Tel Aviv University, Israel
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Zlotogorski-Hurvitz, A., Dayan, D., Chaushu, G. et al. Morphological and molecular features of oral fluid-derived exosomes: oral cancer patients versus healthy individuals. J Cancer Res Clin Oncol 142, 101–110 (2016). https://doi.org/10.1007/s00432-015-2005-3
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DOI: https://doi.org/10.1007/s00432-015-2005-3