MicroRNA-146a and microRNA-155 show tissue-dependent expression in dental pulp, gingival and periodontal ligament fibroblasts in vitro

Abstract

MicroRNAs (miRNAs) are small noncoding RNAs showing a tissue-specific expression pattern, and whose function is to suppress protein synthesis. In this study, we hypothesized that expression of miRNAs would differ among fibroblasts from dental pulp (DPF), gingiva (GF) and periodontal ligament (PLF) in vitro. Once established by an explant technique, DPF, GF and PLF were collected for RNA isolation and subjected to a miRNA microarray. Next, cells were stimulated with E. coli lipopolysaccharide (LPS) for 24 h and then collected for RNA isolation. Expression of miR-146a and miR-155 was investigated by qPCR. Microarray screening revealed several miRNAs that showed specifically high expression in at least one of the fibroblast subtypes. These molecules are potentially involved in the regulation of extracellular matrix turnover and production of inflammatory mediators. Microarray analysis showed that both miR-146a and miR-155 were among the miRNAs expressed exclusively in GF. qPCR demonstrated significant upregulation of miR-146a only in GF after LPS stimulation, whereas basal expression of miR-155 was higher in GF than in the other cell subtypes. LPS downregulated the expression of miR-155 only in GF. Our results suggest that the expression and regulation of miR-146a and miR-155 are more pronounced in GF than in DPF and PLF. (J Oral Sci 56, 157-164, 2014)

Angiotensin II Type 1 Receptor Knockdown Impairs Interleukin‐1β‐Induced Cytokines in Human Periodontal Fibroblasts

“…This result highlights the hypothesis that some post‐transcriptional modification can impact final protein production by these cells. A previous study showed that micro‐RNA‐146a and micro‐RNA‐155 reveal tissue‐dependent expression comparing HGF and HPLF, 43 which could be a possible explanation for differences observed in expression of cytokines by these two fibroblast subtypes.…”

The Role of c‐SKI in Regulation of TGFβ‐Induced Human Cardiac Fibroblast Proliferation and ECM Protein Expression

“…A, Venn diagram generated by http://bioinformatics.psb.ugent.be/webtools/Venn/). Combined with previous studies, 15 candidate upstream miRNAs which are related to ECM were chosen for further assays: miR‐17 [Yu et al, ], miR‐449a [Han et al, ], miR‐33a [Li et al, ], miR‐15b [Kang et al, ], miR‐29c [Chuang et al, ], miR‐25 [Divakaran et al, ], miR‐155 [Sipert et al, ], miR‐24 [Wang et al, ], miR‐29b [Wang et al, ], miR‐29a [James et al, ], miR‐23b [Zhao et al, ], miR‐34a [Lichner et al, ], miR‐17 [Liu et al, ], miR‐204 [Chen et al, ], and miR‐21 [Rubis et al, ] (Fig. A).…”

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“…31 According to the previous position paper, de-fining MSCs needed to meet the following three minimal criteria 32 : the expression and regulation of miR-146a and miR-155 are more significant in gingiva than in other tissues. 17 According to this study, we checked the expression of two gingiva-dependent miRNAs:…”

“…After evaluation, cDNA was amplified using the PCR primers and Universal Master Mix II (Applied Biosystems) in a PCR instrument (ABI Prism 7300 Sequence Detection System, Applied Biosystems). According to the manufacturer's instructions, cycling parameters were showed in supporting information Table …”