Functional analysis of miRNAs combined with TGF-β1/Smad3 inhibitor in an intrauterine rat adhesion cell model.
Mol Cell Biochem, 2020/7;470(1-2):15-28.
Liu S[1], Huang X[1], Liu Y[1], Song D[1], Xiao Y[2]
Affiliations
PMID: 32447720DOI: 10.1007/s11010-020-03741-7
Impact factor: 3.842
Abstract
In this study, we aimed to study the role of miRNAs in intrauterine adhesion (IUA) disease. An IUA cell model was constructed by TGF-β1. Smad3 inhibitor (SIS3) can inhibit the Smad3 signaling pathway and affect the role of TGF-β1; thus, it was used to identify the role of Smad3 and related miRNAs in IUA. Cell number significantly increased in the TGF-β1 group after 72 h and 96 h, respectively, compared with that in the control group (P < 0.05). However, cell proliferation was significantly decreased in the TGF-β1 + SIS3 group (P < 0.0001). Cell apoptosis was increased in the TGF-β1 + SIS3 group compared with that in the TGF-β1 group. Western Blot (WB) analysis suggested that TGF-β1 treatment could effectively increase the expression of α-SMA, COL1, Smad3, and p-Smad3, which could be inhibited by SIS3 treatment. A total of 235 and 530 differentially expressed miRNAs in the TGF-β1 + SIS3 group were significantly up- and downregulated compared with those in the TGF-β1 group, respectively. These differentially expressed miRNAs were enriched in the MAPK and PI3K-AKT pathways. The ten most differentially expressed miRNAs were selected to verify their expressions using quantitative real-time polymerase chain reaction (qPCR). Furthermore, overexpression of rno-miR-3586-3p and rno-miR-455-5p can promote cell proliferation and exacerbate the IUA pathogenic process. However, overexpression of rno-miR-204-3p and rno-miR-3578 can inhibit cell behavior and IUA progression. The above results can provide detailed information for the understanding of IUA molecular mechanisms.
Keywords: Cell behavior; Intrauterine adhesion; Smad3 inhibitor; TGF-β1; miRNAs
MeSH terms
Actins; Animals; Apoptosis; Cell Adhesion; Cell Proliferation; Cells, Cultured; Collagen Type I; Female; MAP Kinase Signaling System; MicroRNAs; Rats; Rats, Sprague-Dawley; Signal Transduction; Smad3 Protein; Tissue Adhesions; Transforming Growth Factor beta1; Uterus
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