Anti-inflammatory role of microRNA-429 in human gingival epithelial cells-inhibition of IL-8 production through direct binding to IKKβ mRNA
Anti-inflammatory role of microRNA-429 in human gingival epithelial cells-inhibition of IL-8 production through direct binding to IKKβ mRNA
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DOI：
10.3892/mmr.2021.12220
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发表时间：
2021-08-01
影响因子：
3.4
通讯作者：
Amano, Hirotaka
中科院分区：
文献类型：
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作者：
Kawasaki, Hiromichi;Amano, Hirotaka
microRNAs（miRNAs）是一类小分子非编码RNA，通过调控多种基因的表达，在炎症反应中起着重要作用。然而，miRNAs调控口腔上皮细胞炎症相关分子的分子机制仍有待阐明。本研究通过在牙龈鳞状细胞癌细胞系Ca 9 -22细胞中进行miR-429的获得/丧失功能研究来检查miR-429的生物学功能，所述细胞通过分别用miR-429模拟物或miR-429抑制剂瞬时转染而过表达或低表达miR-429。结果表明，miR-429的过表达抑制了几种白细胞介素（包括IL-8）的mRNA水平。此外，在基础和TNF-α刺激条件下，miR-429的过表达降低了IL-8的分泌，而当miR-429表达不足时，IL-8的分泌增强。miR-429的过表达抑制了转录因子NF-κ B B的激活。此外，我们发现miR-429通过直接结合IKK β mRNA的3 '-非翻译区抑制IKK β的mRNA和蛋白水平。此外，通过小干扰RNA下调IKK β可降低Ca 9 -22细胞中NF-κ B活性和IL-8的产生。综上所述，研究结果揭示了miR-429调节牙龈细胞中炎症介质的分子机制，并表明其可用作口腔炎症性疾病的治疗靶点。
MicroRNAs (miRNAs), a family of small non-coding RNAs, serve a pivotal role in the regulation of the inflammation by modulating the expression of various genes. However, the molecular mechanism by which miRNAs regulate inflammation-associated molecules in oral epithelial cells remains to be elucidated. The present study examined the biological function of miR-429 by performing the gain-/loss-of-function studies of miR-429 in a gingival squamous cell carcinoma line Ca9-22 cells that either over- or under-expressed miR-429 through transient transfection with miR-429 mimic or miR-429 inhibitor, respectively. The results demonstrated that the over-expression of miR-429 suppressed the mRNA level of several interleukins, including IL-8. In addition, the over-expression of miR-429 reduced IL-8 secretion under the basal and TNF-alpha stimulated conditions, whereas the secretion of IL-8 was enhanced when miR-429 was under-expressed. The over-expression of miR-429 inhibited the activation of the transcription factor NF-kappa B. Furthermore, we found that miR-429 suppressed both mRNA and protein levels of IKK beta via its direct binding to the 3 '-untranslated region of IKK beta mRNA. In addition, the downregulation of IKK beta by small interfering RNA reduced both NF-kB activity and IL-8 production in Ca9-22 cells. Taken together, the findings revealed the molecular mechanism of miR-429 to regulate the inflammatory mediator in gingival cells and suggested that it could be useful as a therapeutic target of oral inflammatory diseases.