FUT8 modulates galectin-3 expression to regulate TGF-β1-mediated fibrosis of lung fibroblasts.
10.12122/j.issn.1673-4254.2022.08.08
- Author:
Wei Wei GAO
1
;
Dai Jian LIU
1
;
Xiao Meng ZHANG
1
;
Qing Qing FENG
1
;
Ying LIU
1
Author Information
1. Department of Respiratory and Critical Care Medicine, Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China.
- Publication Type:Journal Article
- Keywords:
FUT8;
fibrosis;
galectin-3;
migration;
proliferation
- MeSH:
Animals;
Bleomycin/metabolism*;
Fibroblasts/metabolism*;
Fibrosis;
Fucosyltransferases/metabolism*;
Galectin 3/genetics*;
Humans;
Lung/metabolism*;
Mice;
Proto-Oncogene Proteins c-akt/metabolism*;
Transforming Growth Factor beta1/metabolism*
- From:
Journal of Southern Medical University
2022;42(8):1166-1173
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the regulatory role of α-1, 6-fucosyltransferase (FUT8) in TGF-β1-induced proliferation, migration and fibrosis of human embryonic lung fibroblasts (MRC-5 cells) and explore the underlying molecular mechanism.
METHODS:C57/BL6 mice were randomized into 4 groups for treatment with saline (control group), bleomycin, bleomycin+sh-NC or bleomycin+sh-FUT8, and pulmonary fibrosis was observed using Masson staining.MRC-5 cells were transfected with si-NC, FUT8 siRNA (si-FUT8), or both si-FUT8 and a galectin-3(Gal-3) overexpression plasmid (pcDNA3.1-Gal) prior to TGF-β1 treatment, and the changes in cell proliferation and migration were assessed using CCK-8 assay, BrdU assay, and wound healing assay; the changes in the expression levels of α-SMA, collagen I (COLIA1) and extracellular matrix fibronectin (FN) were detected with real-time quantitative PCR (RT-qPCR) and Western blotting.The interaction of FUT8 and Gal-3 was tested using coimmunoprecipitation (Co-IP) assay, and the effect of FUT8 silencing on Gal-3 and FAK/Akt signaling pathways was analyzed.
RESULTS:FUT8 knockdown significantly reduced bleomycin-induced extracellular collagen deposition in the lung tissues of the mice.Silencing FUT8 obviously inhibited cell proliferation (P < 0.05) and migration mediated by TGF-β1.FUT8 knockdown down-regulated the mRNA and protein levels of α-SMA, COLIA1 and FN (P < 0.05) in the cells.Coimmunoprecipitation analysis showed that FUT8 interacted with Gal-3.Silencing FUT8 significantly down-regulated Gal-3 expression and inhibited the activation of the FAK/Akt signaling pathway (P < 0.05).Overexpression of Gal-3 obviously reversed the effects of FUT8 silencing on cell proliferation, migration and fibrosis (P < 0.05).
CONCLUSION:FUT8 regulates TGF-β1-induced proliferation, migration and fibrosis of MRC-5 cells by modulating Gal-3 expression, in which the FAK/Akt pathway may play a role.
