Difficulties detecting miRNA-203 in human whole saliva by the use of PCR

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Abstract

Objectives: Oral Lichen Planus (OLP) is a chronic disease of the oral mucosa, and according to the WHO also a pre malignant condition. Micro-RNAs are short non coding RNAs capable of regulating mRNA expression. MiRNA:scan be detected in tissue, blood and human whole saliva (HWS) and recently we have shown miR-203 to be up-regulated in tissue from OLP lesions. Study Design: In order to see whether mRNA as well as miR-203 could be detected also in HWS, saliva from healthy controls and patients with OLP were analysed using two different PCR methods. Results: Results showed low mRNA and miRNA levels in general in HWS samples, making it hard to generate conclusive results. Conclusions: In order to make HWS a valuable source for different analyses, more sensitive PCR techniques capable of detecting very low levels of mRNAand miRNAas well as more efficient methods for extraction of RNA are needed.

Key words:miRNA-203, saliva, PCR.

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A skin microRNA promotes differentiation by repressing 'stemness'.

Rui Yi, Matthew Poy, Markus Stoffel … (2008)

In stratified epithelial tissues, homeostasis relies on the self-renewing capacity of stem cells located within the innermost basal layer. As basal cells become suprabasal, they lose proliferative potential and embark on a terminal differentiation programme. Here, we show that microRNA-203 is induced in the skin concomitantly with stratification and differentiation. By altering miR-203's spatiotemporal expression in vivo, we show that miR-203 promotes epidermal differentiation by restricting proliferative potential and inducing cell-cycle exit. We identify p63 as one of the conserved targets of miR-203 across vertebrates. Notably, p63 is an essential regulator of stem-cell maintenance in stratified epithelial tissues. We show that miR-203 directly represses the expression of p63: it fails to switch off suprabasally when either Dicer1 or miR-203 is absent and it becomes repressed basally when miR-203 is prematurely expressed. Our findings suggest that miR-203 defines a molecular boundary between proliferative basal progenitors and terminally differentiating suprabasal cells, ensuring proper identity of neighbouring layers.

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Oral lichen planus and oral lichenoid lesions: diagnostic and therapeutic considerations.

Isaäc van der Waal, Palle Holmstrup, Mark Schifter … (2007)

Several therapeutic agents have been investigated for the treatment of oral lichen planus (OLP). Among these are corticosteroids, retinoids, cyclosporine, and phototherapy, in addition to other treatment modalities. A systematic review of clinical trials showed that particularly topical corticosteroids are often effective in the management of symptomatic OLP lichen planus. Systemic corticosteroids should be only considered for severe widespread OLP and for lichen planus involving other mucocutaneous sites. Because of the ongoing controversy in the literature about the possible premalignant character of OLP, periodic follow-up is recommended. There is a spectrum of oral lichen planus-like ("lichenoid") lesions that may confuse the differential diagnosis. These include lichenoid contact lesions, lichenoid drug reactions and lichenoid lesions of graft-versus-host disease. In regard to the approach to oral lichenoid contact lesions the value of patch testing remains controversial. Confirmation of the diagnosis of an oral lichenoid drug reaction may be difficult, since empiric withdrawal of the suspected drug and/or its substitution by an alternative agent may be complicated. Oral lichenoid lesions of graft-versus-host disease (OLL-GVHD) are recognized to have an association with malignancy. Local therapy for these lesions rests in topical agents, predominantly corticosteroids.

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miR-203 represses 'stemness' by repressing DeltaNp63.

Daniel Knight, A M Lena, R Shalom-Feuerstein … (2008)

The epidermis, the outer layer of the skin composed of keratinocytes, is a stratified epithelium that functions as a barrier to protect the organism from dehydration and external insults. The epidermis develops depending on the transcription factor p63, a member of the p53 family of transcription factors. p63 is strongly expressed in the innermost basal layer where epithelial cells with high clonogenic and proliferative capacity reside. Deletion of p63 in mice results in a dramatic loss of all keratinocytes and loss of stratified epithelia, probably due to a premature proliferative rundown of the stem and transient amplifying cells. Here we report that microRNA (miR)-203 is induced in vitro in primary keratinocytes in parallel with differentiation. We found that miR-203 specifically targets human and mouse p63 3'-UTRs and not SOCS-3, despite bioinformatics alignment between miR-203 and SOCS-3 3'-UTR. We also show that miR-203 overexpression in proliferating keratinocytes is not sufficient to induce full epidermal differentiation in vitro. In addition, we demonstrate that miR-203 is downregulated during the epithelial commitment of embryonic stem cells, and that overexpression of miR-203 in rapidly proliferating human primary keratinocytes significantly reduces their clonogenic capacity. The results suggest that miR-203, by regulating the DeltaNp63 expression level, is a key molecule controlling the p63-dependent proliferative potential of epithelial precursor cells both during keratinocyte differentiation and in epithelial development. In addition, we have shown that miR-203 can regulate DeltaNp63 levels upon genotoxic damage in head and neck squamous cell carcinoma cells, thus controlling cell survival.

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Author and article information

Journal

Journal ID (nlm-ta): Med Oral Patol Oral Cir Bucal

Journal ID (iso-abbrev): Med Oral Patol Oral Cir Bucal

Journal ID (publisher-id): Medicina Oral S.L.

Title: Medicina Oral, Patología Oral y Cirugía Bucal

Publisher: Medicina Oral S.L.

ISSN (Print): 1698-4447

ISSN (Electronic): 1698-6946

Publication date (Print): March 2015

Publication date (Electronic): 5 December 2014

Volume: 20

Issue: 2

Pages: e130-e134

Affiliations

[1 ]Department of Odontology, Umeå University, SE-901 85 Umeå, Sweden

[2 ]Department of Medical Biosciences, Umeå University, SE-901 85 Umeå, Sweden

Author notes

Department of Odontology Umeå University SE-901 85 Umeå, Sweden , E-mail: karin.danielsson@ 123456odont.umu.se

Conflict of interest statement: The authors have declared that no conflict of interest exist.

Article

Publisher ID: 20172

DOI: 10.4317/medoral.20172

PMC ID: 4393973

PubMed ID: 25475777

SO-VID: b707c27b-981e-442f-9142-a43c8930bb10

License:

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Difficulties detecting miRNA-203 in human whole saliva by the use of PCR

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Most cited references 14

A skin microRNA promotes differentiation by repressing 'stemness'.

Oral lichen planus and oral lichenoid lesions: diagnostic and therapeutic considerations.

miR-203 represses 'stemness' by repressing DeltaNp63.

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