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目的探讨转录因子C/EBP-β调节人子宫内膜基质细胞蜕膜化的作用机制。方法采用8-Br-c AMP和MPA方法对人子宫内膜基质细胞体外蜕膜化实施处理分析,并通过瞬时转染方法调节转录因子C/EBP-β表达,然后分别收集蜕膜化处理不同时间及干扰前后的细胞,利用流式细胞仪检测细胞蜕膜化不同时间细胞分期的改变情况以及干扰前后细胞周期的变化,并采用Western blot和荧光定量实时PCR法(RT-PCR)分别在mRNA水平和蛋白质水平下检测细胞周期蛋白p57在转染siRNA组和对照组中的表达情况。结果子宫内膜基质细胞在体外蜕膜化中S期细胞的比例随着细胞蜕膜化时间的增长而减少,而G0/G1期细胞的比例随着细胞蜕膜化时间的增长呈现明显的上升趋势,G2/M期细胞的比例随着细胞蜕膜化时间的增长而显著下降,差异均有统计学意义(P<0.05);而转染C/EBP-βsiRNA后再蜕膜化48 h后的S期细胞比例变化范围为4.28%~6.88%,差异有统计学意义(P<0.05);G0/G1期细胞比例范围为86.07%~87.34%,G2/M期细胞比例范围为7.04%~8.38%,均呈逐渐下降趋势,但差异无统计学意义(P>0.05);且转染C/EBP-βsiRNA以后,对人子宫内膜基质细胞中细胞周期蛋白p57的表达进行检测分析,发现p57的表达有明显的下降趋势,差异有统计学意义(P<0.05)。结论转录因子C/EBP-β在人子宫内膜基质细胞蜕膜化过程中通过调节细胞周期蛋白p57使细胞脱离其周期而分化为一种新的蜕膜细胞,在蜕膜化过程中发挥重要作用。
Objective To investigate the mechanism of transcription factor C / EBP-β in regulating the decidualization of human endometrial stromal cells. Methods 8-Br-c AMP and MPA methods were used to analyze the decidualization of human endometrial stromal cells in vitro and the expression of C / EBP-β was regulated by transient transfection. The decidualization was then separately collected Time and interference before and after the cells, the use of flow cytometry cell decidualization at different stages of cell staging changes and changes in cell cycle before and after interference, and using Western blot and real-time quantitative PCR (RT-PCR) mRNA The expression of cyclin-p57 in transfected siRNA group and control group was detected at the level and protein level. Results In endometrial stromal cells, the proportion of S phase cells in endometrial stromal cells decreased with the time of decidualization, while the proportion of G0 / G1 phase cells increased markedly with the increase of decidualization time (P <0.05). However, the percentage of cells in G2 / M phase decreased significantly with the increase of cell decidualization time (P <0.05), while the expression of C / EBP- (P <0.05). The proportion of cells in G0 / G1 phase ranged from 86.07% to 87.34%, the proportion of cells in G2 / M phase ranged from 7.04% to 6.88%, the difference was statistically significant 8.38%, all showed a gradual downward trend, but the difference was not statistically significant (P> 0.05). After transfection of C / EBP-βsiRNA, the expression of cyclin p57 in human endometrial stromal cells was detected and analyzed. The expression of p57 had a significant downward trend, the difference was statistically significant (P <0.05). Conclusion Transcription factor C / EBP-β can differentiate into a new type of decidual cells during the process of decidualization by regulating the expression of cyclin P57 during the decidualization of human endometrial stromal cells effect.