Expression of DACT1 in children with asthma and its regulation mechanism

Abstract

Abstract. The aim of the present study was to detect DACT1 expression levels in the lungs of children with asthma, and to investigate its role and molecular mechanisms in regulating the expression of inflammatory factors in RAW264.7 cells. DACT1, DACT2 and DACT3 expression was analyzed in biopsy specimens from 10 cases of newly diagnosed children with asthma and 10 healthy controls by reverse transcription-quantitative polymerase chain reaction, and their expression was confirmed in RAW264.7 cells. DACT1 expression was silenced by small interfering RNA or enhanced by transfection of pcDNA-3.1-DACT1 in RAW264.7 cells, and expression of β-catenin and inflammatory factors, interleukin (IL) 5, IL6 and IL13, was analyzed. Nuclear translocation of β-catenin was detected by western blot analysis, and the effect of DACT1 on β-catenin was investigated with rescue experiments. Regulation of the Wnt signaling pathway by DACT1 and β-catenin was analyzed in RAW264.7 cells after recombinant Wnt5A stimulation. DACT1, DACT2 and DACT3 were significantly upregulated in specimens from children with asthma compared with controls (P<0.05) and the expression of DACT1 was significantly more increased compared with DACT2 and DACT3 (P<0.05). Inhibition of DACT1 expression significantly suppressed IL5, IL6 and IL13 mRNA expression levels compared with the control (P<0.05), while upregulated DACT1 expression significantly increased IL5, IL6 and IL13 mRNA expression (P<0.05). DACT1 inhibited the expression and nuclear translocation of β-catenin, while overexpression of β-catenin significantly inhibited the biological function of DACT1 (P<0.05). Overexpression of β-catenin also significantly suppressed the upregulation of IL5, IL6 and IL13 mRNA induced by pcDNA3.1-DACT1 transfection (P<0.05). Following the addition of Wnt5A, overexpression of DACT1 inhibited the expression and nuclear translocation of β-catenin, and upregulated IL5, IL6 and IL13 mRNA expression. In conclusion, DACT1 was indicated to be upregulated in lung tissues from children with asthma, which could induce higher pro-inflammatory factor expression. DACT1 may act via inhibiting the expression and nuclear translocation of β-catenin, a factor in the Wnt signaling pathway. The present results suggested that DACT1 may be a potential target for the treatment of asthma.

Human amniotic membrane mesenchymal stem cell‐conditioned medium reduces inflammatory factors and fibrosis in ovalbumin‐induced asthma in mice

“…As shown in Figure 2 One of the most important Th2 cytokines involved in asthma is IL-4, which plays an important role in promotion of the remodelling process (Zhang, Yang, Chen, Liu, & Yuan, 2018). Interleukin-4 induces differentiation of fibroblasts to myofibroblast and plays a role in the synthesis and deposition of collagen under the basement membrane and, consequently, thickening of the basement membrane and subepithelial fibrosis (Tagaya & Tamaoki, 2007).…”

Transcriptomic analysis of TGFβ-mediated fibrosis in primary human Tenon’s fibroblasts

“…DACT1 was upregulated in the TGFβ sample; it has been shown to inhibit the Wnt signalling pathway. Therefore, DACT1 may release inflammatory factors that were previously suppressed by Wnt and promote local inflammation 33 . Of the Wnt signalling pathways that were highly enriched in our gene set, the Wnt/Ca 2+ pathway (Figure 7b) is known to play a critical role in profibrotic and proinflammatory processes including actin polymerisation, cell adhesion, cell migration and NFAT signalling 34 .…”

Transcriptomic analysis of TGFβ-mediated fibrosis in primary human Tenon’s fibroblasts

“…DACT1 was upregulated in the TGFβ sample; it has been shown to inhibit the Wnt signalling pathway. Therefore, DACT1 may release in ammatory factors that were previously suppressed by Wnt and promote local in ammation 33 . Of the Wnt signalling pathways that were highly enriched in our gene set, the Wnt/Ca 2+ pathway (Fig.…”