Curdione inhibits the proliferation of human uterine leiomyomas by targeting YTHDF1.

BACKGROUND

Curdione, an active sesquiterpene found in Curcumae Rhizoma, exhibits strong anti-inflammatory and antitumor properties, but its mechanism of action against uterine leiomyomas (ULMs) is not yet clear.

PURPOSE: This study aims to clarify the anti-proliferative effects of curdione on ULMs and explore the molecular mechanisms, with specific focus on the role of the m6A reader protein YTHDF1.

METHODS

To evaluate the effects of curdione on human ULM cells, assays such as CCK-8, crystal violet staining, EdU, TUNEL, and Annexin V-FITC/PI were used to assess cell proliferation and apoptosis. Key m6A targets were identified through a comprehensive analysis involving GEO data integration, molecular docking, and surface plasmon resonance (SPR). RNA-seq and MeRIP-seq characterized N6-methyladenosine (m6A) methylation patterns and identify downstream regulatory targets. Lentiviral transfection was used to develop YTHDF1 knockdown and overexpression cell models, with target functions validated through molecular assays. The efficacy of curdione was assessed in a rat ULM model using histopathological analysis, electron microscopy, and molecular assays.

RESULTS

Curdione exhibited an inhibitory effect on the viability of ULM cells, significantly reducing proliferation rates and inducing apoptosis. It was observed to downregulate YTHDF1 expression at both transcriptional and translational levels. MeRIP-seq analysis identified reprogramming of m6A methylation, with 231 hypermethylated and 984 hypomethylated peaks, alongside a reduction in THBS1 m6A modification and expression. Stable YTHDF1 knockdown and overexpression models were established through lentiviral transfection, confirming YTHDF1 as a pivotal mediator of curdione's anti-proliferative effects. In a rat ULM model, curdione administration resulted in a decreased uterine index, improved histopathological features, reduced collagen deposition, and normalization of serum inflammatory cytokines and sex hormones. Immunofluorescence and western blot analyses verified the co-localization and coordinated downregulation of YTHDF1 and THBS1 in uterine tissue.

CONCLUSION

Curdione exerts its effects by downregulating YTHDF1, thereby influencing m6A modification and the translation of THBS1 mRNA. This study highlights curdione's potential as a therapeutic agent for ULMs and suggests that targeting YTHDF1 could be an effective management strategy.