YY1通过转化生长因子β1信号轴在滋养细胞功能调控中的探索

Exploration of YY1 in the Regulation of Trophoblast Function Through TGF-β1 Signal Axis

目的:探讨Yin Yang-1(YY1)-转化生长因子β1(TGF-β1)-基质金属蛋白酶(MMPs)信号轴对滋养层细胞活性、侵袭和迁移能力的影响。方法:以HTR8/SVneo滋养细胞系为研究对象,构建YY1敲低(siYY1)和正常表达(阴性对照,NC)的细胞模型,实验分为NC组、siYY1组和siYY1+SD-208(TGF-β抑制剂,1μmol/L,48 h)组。采用实时荧光定量聚合酶链反应(PCR)和蛋白质印迹(Western blotting)检测信号通路相关分子,CCK-8法检测细胞活性,划痕实验和Trans-well实验检测细胞迁移和侵袭能力。结果:siYY1组细胞活性、细胞迁移和侵袭能力都明显减弱,MMP-2和MMP-9表达水平显著降低,而基质金属蛋白酶组织抑制物1(TIMP1)和TIMP2表达水平都明显升高。此外,TGF-β1、Smad2和Smad3的表达水平都显著增高,而抑制性Smad7的表达水平却降低。随后用TGF-β抑制剂SD-208处理后,能够显著改善YY1敲低组细胞的活性、侵袭和迁移能力。结论:YY1-TGF-β1信号轴能调控滋养层细胞活性、侵袭和迁移能力,可能参与胚胎的着床和胎盘发育,致使流产的发生。

Objective: To investigate the effects of YY1-TGF-β1-MMPs signal axis on trophoblast cell activity, invasion and migration. Methods: HTR8/SVneo trophoblast cell line was used as the research object to construct the cell model of the YY1 knockdown(siYY1) group and the normal expression(NC) group. The experimental groups were divided into NC group, si YY1 group and si YY1+SD-208(TGF-β inhibitor, 1 μmol/L, 48 h) group. RT-PCR and Western blotting were used to detect the expression of the signaling pathway related molecules;CCK-8 was used to detect cell viability;scratch assay and Trans-well assay were used to detect cell migration and invasion. Results: The cell viability, migration and invasion ability of si YY1 group were significantly attenuated, and the expression levels of MMP-2 and MMP-9 were significantly decreased, while TIMP1 and TIMP2 were significantly increased. In addition, the expression levels of TGF-β1, Smad2 and Smad3 were significantly increased, while the expression of inhibitory Smad7 was decreased. Subsequent treatment with TGF-β inhibitors significantly improved the activity,invasion and migration of YY1 knockdown cells. Conclusions: The signal axis of YY1-TGF-β1 can regulate the activity, invasion and migration of trophoblast cells, which may be involved in embryo implantation and placental development, resulting in miscarriage.