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Hormone-dependent protection against oxidative stress in uterine leiomyoma

ABSTRACT: Context. Uterine leiomyomas exist in a chronically pro-oxidant environment characterized by reduced antioxidant capacity, yet they persist and grow, suggesting the activation of adaptive stress-tolerance mechanisms. Although estrogen and progesterone are central drivers of leiomyoma growth, their role in regulating mitochondrial function under oxidative stress remains incompletely understood. Objective. To investigate how estrogen and progesterone signaling modulate oxidative stress–induced transcriptional programs and mitochondrial metabolic function in uterine leiomyoma cells. Design. Primary patient-derived leiomyoma spheroids were exposed to paraquat-induced oxidative stress in the presence of estradiol, progestins, or hormone receptor antagonists. Transcriptomic responses were assessed by RNA sequencing followed by differential expression and gene set enrichment analyses. Functional mitochondrial metabolism was evaluated using Seahorse extracellular flux assays, and cellular outcomes including senescence and cell death were quantified. Patients. Uterine leiomyoma tissues were obtained from patients undergoing hysterectomy, and primary patient-derived leiomyoma cells were used to generate three-dimensional spheroid cultures. Results. Estradiol and progesterone modulated oxidative stress–responsive transcriptional programs associated with senescence, apoptosis, and mitochondrial regulation. Hormone receptor antagonism induced the most pronounced transcriptomic disruption and was associated with increased senescence and cell death. Consistent with these findings, Seahorse analyses demonstrated that hormonal signaling preserved mitochondrial maximal respiration and spare respiratory capacity under oxidative stress, whereas hormone blockade impaired mitochondrial reserve capacity without compensatory glycolytic upregulation. Progesterone receptor isoform–specific effects further supported a role for differential PR signaling in regulating mitochondrial adaptability. Conclusions. Estrogen and progesterone promote mitochondrial adaptability in leiomyoma cells, enabling survival under sustained oxidative stress. These findings identify mitochondria as a key mediator of hormone-dependent stress resilience and highlight mitochondrial metabolism as a potential therapeutic vulnerability in leiomyomas.

ORGANISM(S): Homo sapiens

PROVIDER: GSE314406 | GEO | 2026/07/01

REPOSITORIES: GEO

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Project description:Uterine Leiomyoma are the most common benign tumors in women of reproductive age. Studies show that both leiomyoma and surrounding myometrium exist in a state of high oxidative stress, characterized by reactive oxygen species (ROS) production and impaired antioxidant defenses including defects of manganese superoxide dismutase (MnSOD). Oxidative stress has been proposed as a driver of leiomyoma initiation by promoting oxidative DNA damage leading to mutations in MED12 and leiomyoma growth. We hypothesize that chronic oxidative stress in human myometrial cells induces cellular remodeling and genomic alterations that promote adaptation and contribute to early events in leiomyoma development. To test this hypothesis, we generated stable human myometrial cell lines (Myo-hTERT) of wild type, MnSOD68K and mutant MnSOD68Q that impairs MnSOD antioxidant activity thereby imposing sustained oxidative stress. Experiments were performed both in vitro using three-dimensional spheroid cultures and in vivo using xenograft models in immunocompromised mice, with some studies extending up to 10 months to model chronic oxidative stress. Cells were exposed to the oxidative inducer paraquat (PQ) and analyzed using cellular and molecular approaches including spatial transcriptomics and MED12 mutational analysis. Cells expressing MnSOD68Q exhibited increased intracellular ROS and oxidative DNA damage, as indicated by accumulation of 8-hydroxy-deoxyguanosine (8-OHdG). Transcriptomic analysis revealed activation of pathways associated with oxidative stress responses including collagen/extracellular matrix, cellular senescence and IGF/AKT signaling. Xenografts of primary myometrial cells subjected to prolonged oxidative stress demonstrated similar molecular responses and showed an increased frequency of MED12 mutations. These changes were accompanied by modest increases in cell proliferation with minimal induction of cell death. Together, these findings provide mechanistic evidence that sustained oxidative stress can drive molecular and genomic changes in myometrial cells and support a causal role for chronic redox imbalance in the initiation of uterine leiomyomas.

Project description:Purpose:Our goal was to identify and characterize a new molecular target of Bisphenol A（BPA）in uterine leiomyoma cells to better understand how this compound may affect uterine leiomyomas growth and development. Methods: Primary cultured cell lines of uterine leiomyoma were treated with 0.1% DMSO or 10.0 μmol/L BPA for 48 h before RNA-seq and histone 3 lysine 27 acetylation (H3K27ac) ChIP-seq were performed. Integrative analysis of ChIP-seq and RNA-seq data identifies the key transcription factor (TF)，target gene and signaling pathway. To confirm the expression level of TF and target gene in uterine leiomyomas tissues, we analyzed 10 human uterine leiomyoma tissues and the matched adjacent uterine smooth muscle tissues using western blotting, and 96 paired paraffin-embedded human uterine samples by immunohistochemistry(IHC). The ChIP assay used to confirm the combination between the TF and target gene. In order to verify the functions of key TF, lentivirus was used to knock down the expression level and detect the effects on cell proliferation, cell cycle, and cell tumorigenicity. We further conducted Western blot analysis to confirm the whether the downstream signaling pathway is activated. RESULTS: Integrative Analysis of ChIP-seq and RNA-seq data identifies the key transcription factor XBP1and target gene ITGA2. The qPCR and ChIP- qPCR initially verified the sequencing results. XBP1 and ITGA2 were overexpressed in human uterine leiomyoma tissues. The IHC results confirmed that ITGA2 was significantly correlated with XBP1 expression (R = 0.6708, P < 0.001)， and the ChIP assay showed that XBP1 binds to the the ITGA2 predicted promoter region. BPA promoted uterine leiomyoma cells proliferation both in vitro and in vivo and that XBP1 expression levels modulated the cellular response to this endocrine disruptor. BPA upregulated ITGA2 through XBP1 and activated the downstream PI3K-AKT signaling pathway to promote the proliferation of human uterine leiomyoma cells. CONCLUSION: In conclusion, our current work reveals a novel mechanism by which BPA promotes the proliferation in uterine leiomyoma cells. The XBP1 transcription factor regulates and activates the downstream ITGA2/PI3K/AKT pathway. By defining XBP1 as an important regulatory role of BPA in uterine leiomyoma cell proliferation, they provide new insights into the pathogenesis related to the exposure to BPA and other endocrine disruptors acting similarly, and XBP1 may serve as a candidate molecular target for intervention and treatment of uterine leiomyomas.

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Hormone-dependent protection against oxidative stress in uterine leiomyoma

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