23 March 2019 : Laboratory Research
MicroRNA-744 Inhibits Proliferation of Bronchial Epithelial Cells by Regulating Smad3 Pathway via Targeting Transforming Growth Factor-β1 (TGF-β1) in Severe Asthma
Han Huang1AC, Hongxia Lu1CD, Lihong Liang1CD, Yueli Zhi1DE, Beibei Huo1EF, Linlin Wu1CE, Liping Xu1BF, Zhaobo Shen1AB*DOI: 10.12659/MSM.912412
Med Sci Monit 2019; 25:2159-2168
Abstract
BACKGROUND: Bronchial epithelial cells proliferation plays a pivotal role in airway remodeling in children with severe asthma. MicroRNAs (miRNAs) have gained great attention for many diseases, including asthma. The purpose of this study was to explore the mechanisms that underlie miR-744 modulating bronchial epithelial cells proliferation in severe asthma in children.
MATERIAL AND METHODS: Bronchial epithelial cells were isolated from bronchial biopsies of normal controls and asthmatic subjects. miR-744 and transforming growth factor-β1 (TGF-β1) expressions were measured by quantitative reverse transcription PCR (qRT-PCR). Proliferating cell nuclear antigen (PCNA), phosphorylation or total of mothers against decapentaplegic homolog3 (Smad3) and production of Smad anchor for receptor activation (SARA) were measured via Western blot analysis. A link between miR-744 and TGF-β1 was probed by luciferase activity and RNA immunoprecipitation. Cell proliferation was evaluated using the Cell Proliferation Assay Kit.
RESULTS: Severe asthma showed a significantly elevated cell proliferation rate and reduced abundance of miR-744, which in turn inhibited cell proliferation of bronchial epithelial cells. In particular, TGF-β1 might be a candidate target of miR-744, and enrichment of miR-744 lowered the expression of TGF-β1 at mRNA and protein levels. Indeed, overexpression of miR-744 lowered the proliferation rate of bronchial epithelial cells via driving TGF-β1. Moreover, addition of miR-744 limited phosphorylation of Smad3 but reversed SARA protein abundance by regulating TGF-β1.
CONCLUSIONS: The presence of miR-744 repressed bronchial epithelial cells proliferation through mediating the Smad3 pathway by modulating TGF-β1, providing a promising therapeutic approach for epithelial function in severe asthma.
Keywords: Bronchial Arteries, severe acute respiratory syndrome, Transforming Growth Factor beta, Asthma, Bronchi, Child, Phosphorylation, RNA, Messenger, Respiratory Mucosa, Smad3 Protein, Transforming Growth Factor beta1
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