卵泡刺激素对人卵巢颗粒KGN细胞TGF-β信号传导的影响
Effects of follicle stimulating hormone on TGF-β signal transduction in human ovarian granulosa KGN cells
张铭露 1温苗 2杨泽岸 3于思思 4张媛 3吴彦霖3
作者信息
- 1. 中国食品药品检定研究院,北京 102629;河南省药品医疗器械检验院,郑州 450018
- 2. 中国食品药品检定研究院,北京 102629;中国药科大学药学院,南京 210009
- 3. 中国食品药品检定研究院,北京 102629
- 4. 中国食品药品检定研究院,北京 102629;沈阳药科大学生命科学与生物制药学院,沈阳 117004
- 折叠
摘要
目的 利用抗体芯片技术考察卵泡刺激素(FSH)在不同时间上对卵巢颗粒细胞(KGN)转化生长因子β(TGF-β)信号通路的影响.方法 FSH分别刺激KGN细胞3、6、12、24、48、72 h后收集细胞,进行不同时间点的蛋白磷酸化水平差异含量考察,对蛋白磷酸化水平差异进行GO功能注释和KEGG信号通路富集分析.结果 FSH可以介导TGF-β通路的传导,促进颗粒细胞的DNA合成.FSH作用KGN细胞3、12 h后,TGF-β通路下游因子Smad蛋白磷酸化水平升高;GO富集分析发现,磷酸化Smad参与如复合物组装、信号转导等功能.作用24 h和48 h时对丝裂原活化蛋白激酶通路正向调节,其下游因子环AMP依赖性转录因子2、c-Jun、c-Fos等蛋白磷酸化程度增加,表明除了与Smad依赖型TGF-β通路有关,也上调了非Smad依赖型TGF-β通路.12 h后可观察到抑制性I-Smad功能富集,说明其开始对TGF-β通路进行负调控,与R-Smad竞争性结合,阻断TGF-β信号传导;随着FSH作用的时间增加,对TGF-β信号通路的影响也逐渐变小.结论 FSH可以介导KGN细胞内TGF-β信号通路的传导,促进后续的系列反应.
Abstract
Objective Using protein chip technology to investigate the effects of follicle stimulating hormone (FSH) on the transforming growth factor (TGF)-β signaling pathway of ovarian granulosa cells (KGN) at different times. Methods After FSH stimulation of KGN cells for 3,6,12,24,48,and 72 hours,the cells were collected and the differences in protein phosphorylation levels at different time points were investigated. GO functional annotation and KEGG signaling pathway enrichment analysis were performed on the differences in protein phosphorylation levels. Results FSH can mediate the conduction of TGF-β pathway and promote DNA synthesis in granulosa cells. After 3 and 12 hours of FSH action on KGN cells,the phosphorylation level of downstream factor Smad protein in the TGF-pathway increased. GO enrichment analysis revealed that phosphorylated Smad is involved in functions such as complex assembly and signal transduction. At 24 and 48 hours of action,the mitogen-activated protein kinase pathway was positively regulated,and downstream factors such as cAMP-dependent transcription factor 2,c-Jun,and c-Fos showed an increased degree of phosphorylation,indicating that in addition to being related to the Smad dependent TGF-β pathway,the non Smad dependent TGF-β pathway was also upregulated. After 12 hours,the enrichment of inhibitory I-Smad function was observed,indicating that it began to negatively regulate the TGF-β pathway,competitively bind with R-Smad,and block TGF-β signaling. As the duration of FSH action increases,the impact on the TGF-β signaling pathway gradually decreases. Conclusion FSH can mediate the transmission of TGF-β signaling pathway in KGN cells,promoting subsequent series of reactions.
关键词
卵泡刺激素/转化生长因子β信号通路/人卵巢颗粒细胞/基因芯片/丝裂原活化蛋白激酶通路/GO功能注释/KEGG信号通路富集分析/Smad蛋白Key words
Follicle stimulating hormone/Transforming growth factor-β signaling pathway/Human ovarian granulosa cells/Gene chip/Mitogen-activated protein kinases pathway/GO functional annotation/KEGG signaling pathway enrichment analysis/Smad protein引用本文复制引用
出版年
2024
NETL
NSTL
万方数据