Fan Xiao-Rong, Zhu Hou-Ze, Lei Meng-Ying, Jiang Peng-Jun, Zhou Hui, Xia Wei
Maternal and Child Health Hospital of Hubei Province, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430070, China.
Department of Physiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Curr Med Sci. 2025 Sep 5. doi: 10.1007/s11596-025-00110-7.
To investigate the differential expression of microRNA-144-3p in endometrial cells exposed to copper ions in vitro. The specific mechanism by which microRNA-144-3p is involved in Cu-induced damage to the human endometrial epithelial cells (HEECs) was explored.
HEECs were cultured in copper-containing culture medium to simulate changes in the endometrium after copper intrauterine device (Cu-IUD) implantation. Reverse transcription quantitative PCR (RT-qPCR) was used to detect the differential expression of miR-144-3p in HEECs after Cu treatment. MiRNAs, siRNAs and related inhibitors were used to treat HEECs. The expression levels of related downstream genes were then analyzed by RT-qPCR, Western blotting and immunofluorescence to explore the specific mechanism involved.
MiR-144-3p was significantly upregulated in the Cu-treated HEECs. The expression of P-NF-κB, MMP9, TGF-β3 and P-SMAD3 was significantly decreased in HEECs treated with 10 μg/mL Cu. MiR-144-3p regulated the expression of metallothionein 1A (MT1A) and thrombospondin-1 (THBS-1) in Cu-treated HEECs. The expression of P-NF-κB can be regulated by MT1A, and an inhibitor of P-NF-κB can significantly reduce the expression of MMP9 in Cu-treated HEECs. The expression of TGF-β3 can be regulated by THBS-1, and a TGF-β3 inhibitor can significantly reduce the expression of SMAD3 in Cu-treated HEECs. The proliferative capacity of HEECs treated with MMP9 or SMAD3 inhibitors was significantly reduced.
The increased Cu concentration led to the upregulation of miR-144-3p, further reducing the expression levels of its target genes (MT1A and THBS-1), which in turn downregulated the expression of NF-κB, MMP9, TGF-β3 and SMAD3, ultimately leading to increased endometrial cell damage and decreased cell proliferation.
研究体外暴露于铜离子的子宫内膜细胞中微小RNA-144-3p的差异表达。探讨微小RNA-144-3p参与铜诱导的人子宫内膜上皮细胞（HEECs）损伤的具体机制。
将HEECs培养于含铜培养基中，以模拟宫内节育器（Cu-IUD）植入后子宫内膜的变化。采用逆转录定量PCR（RT-qPCR）检测铜处理后HEECs中miR-144-3p的差异表达。使用微小RNA、小干扰RNA及相关抑制剂处理HEECs。然后通过RT-qPCR、蛋白质印迹法和免疫荧光分析相关下游基因的表达水平，以探讨其中涉及的具体机制。
在铜处理的HEECs中，miR-144-3p显著上调。用10μg/mL铜处理的HEECs中，P-NF-κB、MMP9、TGF-β3和P-SMAD3的表达显著降低。miR-144-3p调节铜处理的HEECs中金属硫蛋白1A（MT1A）和血小板反应蛋白-1（THBS-1）的表达。P-NF-κB的表达可由MT1A调节，P-NF-κB抑制剂可显著降低铜处理的HEECs中MMP9的表达。TGF-β3的表达可由THBS-1调节，TGF-β3抑制剂可显著降低铜处理的HEECs中SMAD3的表达。用MMP9或SMAD3抑制剂处理的HEECs的增殖能力显著降低。
铜浓度升高导致miR-144-3p上调，进一步降低其靶基因（MT1A和THBS-1）的表达水平，进而下调NF-κB、MMP9、TGF-β3和SMAD3的表达，最终导致子宫内膜细胞损伤增加和细胞增殖减少。
