MiR-200b attenuates IL-6 production through IKK\u03b2 and ZEB1 in human gingival fibroblasts

Yohei Nakayama,Ayako Kato,Sari Matsui,Natsuko Tanabe,Tomohiro Nakayama,Naoto Suzuki,Hideki Takai,Liming Zhou,Masaru Mezawa,Yorimasa Ogata,Yuki Suzuki,Noriaki Kamio

doi:10.1007/s00011-018-1192-1

Yohei Nakayama, Ayako Kato + Show 10 more

Open Access

https://doi.org/10.1007/s00011-018-1192-1

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Journal: Inflammation Research	Publication Date: Oct 10, 2018
Citations: 21	License type: open-access

Affiliation: Nihon University, Anhui Medical University

Abstract

ObjectiveMicroRNAs (miRNAs) play important roles in biological processes such as cell differentiation, development, infection, immune response, inflammation and tumorigenesis. We previously reported that the expression of miR-200b was significantly increased in inflamed gingiva compared with non-inflamed gingiva. To elucidate the roles of miR-200b in the inflamed gingiva, we have analyzed the effects of miR-200b on the expression of IL-6 in human gingival fibroblasts (HGF).Materials and methodsTotal RNA and protein were extracted from HGF after stimulation by interleukin-1β (IL-1β; 1 ng/ml) or tumor necrosis factor-α (TNF-α; 10 ng/ml) and transfected with miR-200b expression plasmid or miR-200b inhibitor. IL-6, IL-1β, inhibitor of nuclear factor kappa-B kinaseβ (IKKβ), Zinc-finger E-box-binding homeobox 1 (ZEB1) and E-cadherin mRNA and protein levels were analyzed by real-time PCR and Western blot.ResultsIL-1β and TNF-α increased IL-6 mRNA and protein levels, and they were significantly suppressed by miR-200b overexpression, whereas they were further increased by miR-200b inhibitor in HGF. IKKβ and ZEB1 which are target genes of miR-200b negatively regulate E-cadherin. MiR-200b suppressed the expression of IKKβ and ZEB1 and increased E-cadherin mRNA and protein levels in HGF.ConclusionsThese results suggest that miR-200b attenuates inflammatory response via IKKβ and ZEB1 in periodontal tissue.

Highlights

MicroRNAs are single-stranded, small noncoding RNA molecules regulating gene expression by hybridization to targeted transcripts
To determine whether miR-200b is induced by inflammatory cytokines, we stimulated human gingival fibroblasts (HGF) by IL-1β (1 ng/ml), IL-6 (1 ng/ml) and tumor necrosis factor-α (TNF-α) (10 ng/ml) for 24 h and revealed that and they were further significantly increased by miR-200b inhibitor (Fig. 3a, b)
IL-1β and TNF-α induced mRNA and protein levels of IL-6 and mRNA levels of IL-1β, but they were significantly suppressed by miR-200b overexpression in HGF (Fig. 2a–c)

Summary

Introduction

MicroRNAs (miRNAs) are single-stranded, small noncoding RNA molecules regulating gene expression by hybridization to targeted transcripts. They play key roles on biological processes such as cell differentiation, development, infection, immune response, inflammation and tumorigenesis. The role of miRNAs has been extensively studied in the regulation of cellular processes, including proliferation and differentiation, apoptosis, cancer and viral infections. Recent studies have demonstrated the differences in miRNA expression between inflamed and healthy tissues [4–6]. We have previously demonstrated that the three most overexpressed miRNAs in inflamed gingiva from Japanese chronic periodontitis patients were miR-150, miR-223 and miR-200b, and inflammatory cytokines induced miR-150 and miR-223 expressions in human gingival fibroblasts (HGF) [6, 7].

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