Hysteroscopic Centre, Fu Xing Hospital, Capital Medical University, #20 Fuxingmenwai Street, Xicheng District, Beijing, 100038, China.
Mol Cell Biochem. 2020 Jul;470(1-2):15-28. doi: 10.1007/s11010-020-03741-7. Epub 2020 May 23.
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.
在这项研究中，我们旨在研究 miRNA 在宫腔粘连 (IUA) 疾病中的作用。通过 TGF-β1 构建 IUA 细胞模型。Smad3 抑制剂 (SIS3) 可抑制 Smad3 信号通路，影响 TGF-β1 的作用；因此，它被用于鉴定 Smad3 及其相关 miRNA 在 IUA 中的作用。与对照组相比，TGF-β1 组在 72 h 和 96 h 后细胞数量分别显著增加（P < 0.05）。然而，TGF-β1+SIS3 组的细胞增殖明显降低（P < 0.0001）。与 TGF-β1 组相比，TGF-β1+SIS3 组细胞凋亡增加。Western Blot (WB) 分析表明，TGF-β1 处理可有效增加 α-SMA、COL1、Smad3 和 p-Smad3 的表达，而 SIS3 处理可抑制其表达。与 TGF-β1 组相比，TGF-β1+SIS3 组中共有 235 个和 530 个差异表达 miRNA 显著上调和下调，这些差异表达 miRNA 富集在 MAPK 和 PI3K-AKT 通路中。选择十个差异表达最显著的 miRNA 进行定量实时聚合酶链反应（qPCR）验证。此外，rno-miR-3586-3p 和 rno-miR-455-5p 的过表达可促进细胞增殖并加重 IUA 发病过程。然而，rno-miR-204-3p 和 rno-miR-3578 的过表达可抑制细胞行为和 IUA 进展。上述结果可为理解 IUA 分子机制提供详细信息。
