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Background <p>Polycystic ovary syndrome (PCOS) is a complex endocrine disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovaries. Despite its prevalence, the underlying mechanisms contributing to the pathogenesis of PCOS remain poorly understood. This study aims to investigate the gene expression profiles in human granulosa cells and mouse models of PCOS to identify critical risk genes linked to hyperandrogenism and to elucidate their potential roles in the pathogenesis of PCOS. By integrating transcriptomic data from various sources, we seek to uncover the molecular pathways that may contribute to the development and progression of PCOS.</p> Methods <p>We initiated our study by performing RNA sequencing (RNA-seq) on human granulosa cells from patients with polycystic ovary syndrome (PCOS). Subsequently, we induced PCOS-like mouse models by administering dehydroepiandrosterone (DHEA) to 21-day-old mice and conducted RNA-seq on mouse ovaries and granulosa cells to obtain gene expression profiles. Bioinformatics analyses were performed using R software, where we identified differentially expressed genes (DEGs) between PCOS and control groups in both human and mouse samples. These DEGs were analyzed for Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways to determine their biological functions. To identify critical genes involved in PCOS, we integrated DEGs from the three datasets and performed Gene Set Enrichment Analysis (GSEA) to characterize gene expression patterns in enriched pathways. We validated the expression of critical genes using quantitative reverse transcription PCR (RT-qPCR) and Western blot analyses, and determined their localization in mouse granulosa cells through immunofluorescence (IF) and immunohistochemistry (IHC). Finally, we evaluated gene silencing effects of identified genes using siRNA in KGN cell lines.</p> Results <p>From the analysis of the three datasets, we identified seven critical co-genes: CDH23, CLMN, RASEF, ABCB4, PLAU, STEAP4, and CYP2S1. RT-qPCR and Western blot analyses confirmed that PLAU was significantly upregulated in PCOS granulosa cells, with IHC and IF studies showing its localization in the membrane and nucleus. Finally, silencing PLAU in KGN cells resulted in decreased mRNA and protein levels of CYP11A1, CYP19A1, STAR, p65, p-p65, p-IKB, Bax, Bax/Bcl2 and cleaved caspase 3.</p> Conclusion <p>This study identified seven critical genes (CDH23, CLMN, RASEF, ABCB4, PLAU, STEAP4, CYP2S1) expressed in granulosa cells. Notably, we found that the upregulation of PLAU in granulosa cells reduces estrogen secretion and promotes apoptosis by activating the NF-κB signaling pathway in PCOS. This represents the first exploration of PLAU’s involvement in the pathogenesis of PCOS, suggesting that PLAU may serve as a novel molecular target for enhancing granulosa cell function. These findings could provide new insights into diagnostic and therapeutic strategies for PCOS.</p>

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Upregulation of PLAU in granulosa cells disrupts steroid hormone synthesis and promotes apoptosis by activating NF-κB signaling pathway in PCOS

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月29日

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  • Wenxiu Chen,
  • Hanzhi Zhang,
  • Mingxin Jiang,
  • Xing Su,
  • Jingfei Chen,
  • Jianlin Chen

摘要

Background

Polycystic ovary syndrome (PCOS) is a complex endocrine disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovaries. Despite its prevalence, the underlying mechanisms contributing to the pathogenesis of PCOS remain poorly understood. This study aims to investigate the gene expression profiles in human granulosa cells and mouse models of PCOS to identify critical risk genes linked to hyperandrogenism and to elucidate their potential roles in the pathogenesis of PCOS. By integrating transcriptomic data from various sources, we seek to uncover the molecular pathways that may contribute to the development and progression of PCOS.

Methods

We initiated our study by performing RNA sequencing (RNA-seq) on human granulosa cells from patients with polycystic ovary syndrome (PCOS). Subsequently, we induced PCOS-like mouse models by administering dehydroepiandrosterone (DHEA) to 21-day-old mice and conducted RNA-seq on mouse ovaries and granulosa cells to obtain gene expression profiles. Bioinformatics analyses were performed using R software, where we identified differentially expressed genes (DEGs) between PCOS and control groups in both human and mouse samples. These DEGs were analyzed for Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways to determine their biological functions. To identify critical genes involved in PCOS, we integrated DEGs from the three datasets and performed Gene Set Enrichment Analysis (GSEA) to characterize gene expression patterns in enriched pathways. We validated the expression of critical genes using quantitative reverse transcription PCR (RT-qPCR) and Western blot analyses, and determined their localization in mouse granulosa cells through immunofluorescence (IF) and immunohistochemistry (IHC). Finally, we evaluated gene silencing effects of identified genes using siRNA in KGN cell lines.

Results

From the analysis of the three datasets, we identified seven critical co-genes: CDH23, CLMN, RASEF, ABCB4, PLAU, STEAP4, and CYP2S1. RT-qPCR and Western blot analyses confirmed that PLAU was significantly upregulated in PCOS granulosa cells, with IHC and IF studies showing its localization in the membrane and nucleus. Finally, silencing PLAU in KGN cells resulted in decreased mRNA and protein levels of CYP11A1, CYP19A1, STAR, p65, p-p65, p-IKB, Bax, Bax/Bcl2 and cleaved caspase 3.

Conclusion

This study identified seven critical genes (CDH23, CLMN, RASEF, ABCB4, PLAU, STEAP4, CYP2S1) expressed in granulosa cells. Notably, we found that the upregulation of PLAU in granulosa cells reduces estrogen secretion and promotes apoptosis by activating the NF-κB signaling pathway in PCOS. This represents the first exploration of PLAU’s involvement in the pathogenesis of PCOS, suggesting that PLAU may serve as a novel molecular target for enhancing granulosa cell function. These findings could provide new insights into diagnostic and therapeutic strategies for PCOS.