Effect of Galectin-1 on the function of endometrial epithelial cells with different receptivity
10.3760/cma.j.cn101441-20210628-00290
- VernacularTitle:Galectin-1调节子宫内膜容受性的机制研究
- Author:
Liangyu PENG
1
;
Jing YANG
Author Information
1. 武汉大学人民医院产科 430000
- Publication Type:Journal Article
- Keywords:
Galectin-1;
Endometrial receptivity;
WNT pathway;
Epithelial-mesenchymal transition related-protein;
Trophoblast
- From:
Chinese Journal of Reproduction and Contraception
2021;41(11):991-1001
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the expression of Galectin-1 in human ectopic endometrial cells with different receptivity and its role in the process of embryo implantation.Methods:The expression and localization of Galectin-1 in high tolerance endometrial cell RL-95-2 and low tolerance endometrial cell HEC-1-A were detected by real-time quantitative PCR (RT-qPCR), Western blotting and immunofluorescence assay. The invasion and migration of the two kinds of cells were detected by Transwell assay and scratch assay. JAR cells were used to mimic embryos and to detect the migration rate. The expression levels of Galectin-1, epithelial-mesenchymal transition (EMT)-related proteins and WNT/β-catenin signaling pathway-related proteins were analyzed by transfecting Galection-1 siRNA and Galection-1 overexpression vector in the RL-95-2 cells and HEC-1-A cells, respectively. Then the transfected endometrial cells were added with WNT pathway inhibitor DKK1 and activator LiCl respectively, and the changes of EMT-related proteins and the invasion, migration of cells were examined.Results:1) RT-qPCR and Western blotting results showed that Galectin-1 was significantly higher in RL-95-2 than in HEC-1-A ( P=0.020, P=0.030); immunofluorescence experiments showed that Galectin-1 was mainly expressed in the cytoplasm of RL-95-2, while in HEC-1-A cells, Galectin-1 was mainly expressed in the nucleus. 2) The results of cell adhesion assay showed that the adhesion rate of JAR cells to RL-95-2 cells was significantly higher than that to HEC-1-A cells ( P=0.010). 3) After Galectin-1 overexpression, the expression of E-cadherin protein was decreased in the HEC-1-A cells ( P=0.001), and the expressions of N-cadherin and Vimentin were increased ( P=0.003, P=0.023); the exprssioins of β-catenin, WNT4a, WNT5a, and WNT7a were increased ( P=0.025, P=0.004, P=0.005, P=0.001), and the migration ability and invasion ability of HEC-1-A cells were significantly enhanced ( P=0.022, P=0.003). 4) After DKK1 treatment of HEC-1-A cells overexpressing Galectin-1, E-cadherin protein expression was decreased ( P=0.003), N-cadherin and Vimentin protein expressions were increased ( P=0.015, P=0.033), and the migratory ability as well as invasive ability of HEC-1-A cells were significantly increased ( P=0.030, P=0.040). 5) After down-regulation of Galectin-1 expression in RL-95-2 cells, E-cadherin protein expression was increased ( P=0.004), N-cadherin and Vimentin protein expressions were significantly decreased ( P=0.030, P=0.023), and β-catenin, WNT4a, WNT5a, and WNT7a protein expressions were reduced ( P=0.001, P=0.005, P=0.023, P=0.020). 6) After LiCl treatment of RL-95-2 cells with down-regulated Galectin-1 expression, E-cadherin protein expression was increased, and N-cadherin protein and Vimentin protein expressions were decreased compared with LiCl-treated RL-95-2 cells ( P=0.012, P=0.035, P=0.020); RL-95-2 cells migration rate, and invasion number were decreased ( P=0.040, P=0.020). Conclusion:The expression level of Galectin-1 was increased in highly receptivity endometrium compared with low receptivity endometrium. Galectin-1 may regulate the expression of EMT-related proteins through WNT/β-catenin signaling pathway, thus affecting embryo implantation.
