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Oral swirl samples – a robust source of microRNA protected by extracellular vesicles
Author(s) -
Yap T,
Vella LJ,
Seers C,
Nastri A,
Reynolds E,
Cirillo N,
McCullough M
Publication year - 2017
Publication title -
oral diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.953
H-Index - 87
eISSN - 1601-0825
pISSN - 1354-523X
DOI - 10.1111/odi.12603
Subject(s) - microrna , extracellular vesicle , extracellular , microvesicles , saliva , extracellular vesicles , vesicle , rna , biology , chemistry , nucleic acid , microbiology and biotechnology , biochemistry , gene , membrane
Background Micro RNA s are small non‐coding RNA s which are dysregulated in disease states, such as oral cancer. Extracellular vesicles, a potential source of micro RNA , are found in saliva. Objective To demonstrate that a quantifiable amount of micro RNA can be isolated from oral swirl samples. Additionally, we hypothesized that extracellular vesicles may protect contained micro RNA from degradation in these samples. Method A polyethylene glycol‐based precipitation was used for extracellular vesicle enrichment of oral swirl samples. Comparison was made between samples treated with and without RN ase. Further, samples from three subjects were exposed to a range of conditions over 7 days and assessed for presence of micro RNA by reverse‐transcription quantitative PCR . Extracellular vesicles from samples were identified under transmission electron microscopy. Results An adequate quantity of micro RNA for qPCR analysis was extractable from samples despite exposure to conditions under which degradation of RNA would be expected. Conclusion A technique was developed to isolate an adequate quantity of micro RNA for analysis from oral swirl samples. Extracellular vesicle‐associated micro RNA may be protected from degradation. This technique moves towards chairside application of translational micro RNA research in the field of oral cancer prognostics.