Effects of follicle stimulating hormone on TGF-β signal transduction in human ovarian granulosa KGN cells
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.
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