Label-Free Quantitative Proteomic Study of The Effect of Dihydrotestosterone (DHT) On Rat Ovarian GCs

Abstract

Dihydrotestosterone (DHT) is a main androgen in the human body. Previous reports have shown that DHT can affect the proliferation, apoptosis and estrogen and progesterone secretion of ovarian granulosa cells (GCs). An imbalance in DHT secretion leads to GC dysfunction and follicular development disorder.Therefore, exploring the influence of DHT on GCs is necessary. The purpose of this study was to analyze the effect of DHT on GCs through label-free quantitative proteomics (LFQP). After primary cultured rat GCs were treated with DHT (10-8 mol/L), the effect of DHT on GCs was analyzed by LFQP, and some of the differentially expressed proteins (DEPs) were verified by western blotting.A total of 6124.0 proteins were identified, of which 4496.0 were quantifiable. Compared with the control group, 28 proteins were upregulated and 10 were downregulated after DHT intervention. The subcellular localization of DEPs indicates that DHT is involved in the proliferation, migration, molding and metabolism of GCs. Gene Ontology (GO) revealed that DHT downregulated the oxygen transport capacity and oxygen-binding protein of GCs. Orthologous Groups of proteins (COG/KOG) showed that DHT had an important effect on the survival, growth and apoptosis of GCs. Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed that DHT promotes metabolism, amino acid degradation, chemical carcinogenesis, platelet activation and vasoconstriction in GCs. The western blot results were consistent with the proteomics results. Mark3 and Mre11a are DEPs that were upregulated, and Fth1 and Nqo1 were downregulated, which indicated that DHT could promote the proliferation of GCs.This study comprehensively analyzes the impact of DHT on GCs through LFQP and provides clues for further research.