title: MicroRNA-223 affects IL-6 secretion in mast cells via the IGF1R/PI3K signaling pathway.
abstract: In this study, we aimed to assess the effects of microRNA-223 (miR-223) on interleukin-6 (IL-6) secretion in mast cells and determine the underlying molecular mechanisms. Mast cells (P815) were transfected with miR-223 lentiviral vector and miR-223 inhibitor. miR-223 expression was then evaluated using reverse transcription-quantitative PCR (RT-qPCR). IL-6 levels in the supernatant were analyzed using enzyme-linked immunosorbent assay. The signaling pathways in mast cells with downregulated miR-223 were initially evaluated by gene chip. Downregulation of miR-223 and its target gene was tested using a luciferase reporter assay. The expression of phosphate-AKT (p-AKT) and its target protein insulin-like growth factor-1 receptor (IGF1R) was assessed by western blot analysis. Phosphatidylinositol 3-kinase (PI3K)-inhibitor (LY294002) and insulin-like growth factor-1 (IGF1) were used to determine the effect of miR-223 on IL-6 secretion in mast cells. The results showed that microRNA-223 reduced IL-6 concentration in the mast cells. The gene chip results predicted an induction of the PI3K-AKT signaling pathway in the mast cells. Luciferase reporter assay confirmed IGF1R gene to be a target of miR-223. The p-AKT and IGF1R levels increased following miR-223 downregulation in mast cells. In addition, the specific PI3K‑inhibitor LY294002 decreased IL-6 secretion. Incubation with IGF1 resulted in the induction of IL-6 secretion in miR-223‑expressing mast cells. In conclusion, it was shown that miR-223 reduces IL-6 secretion in mast cells by inhibiting the IGF1R/PI3K signaling pathway.
authors.affiliation: Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Qianyuan
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Pediatrics, Jiangsu Huai'an Maternity and Children's Hospital, Huai'an, Jiangsu 223002, P.R. China.
authors.forename: Jin
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Deyu
authors.initials: D
authors.lastname: Zhao
authors.affiliation: Department of Respiratory Medicine, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002867:Chromones / Q000494:pharmacology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D015850:Interleukin-6 / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017526:Receptor, IGF Type 1 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D002867:Chromones; D015850:Interleukin-6; C544877:MIRN223 microRNA, mouse; D035683:MicroRNAs; D009025:Morpholines; D047428:Protein Kinase Inhibitors; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D008156:Luciferases; D058539:Phosphatidylinositol 3-Kinase; D017526:Receptor, IGF Type 1