Triptolide targets JUN to reverse cisplatin resistance of ovarian cancer: insights from single-cell transcriptome analysis and machine learning validation
TL;DR
This study developed a comprehensive strategy integrating computational predictions with in vitro experimental validations that reverses CDDP resistance in PROC by downregulating JUN, which dismantles intracellular pro-survival networks.
Abstract
Background Platinum-resistant ovarian cancer (PROC) is a major clinical challenge driven by profound intratumoral heterogeneity. Triptolide (TP) exhibits promising anti-tumor potential, yet its precise mechanisms within PROC remain elusive due to the limitations of traditional target-screening strategies. Methods This study developed a comprehensive strategy integrating computational predictions with in vitro experimental validations. First, scRNA-seq data were processed to evaluate TP target genes predicted by SwissTargetPrediction, alongside pseudotime trajectory and CellChat intercellular communication analyses. Subsequently, an ensemble of six machine learning (ML) algorithms (LASSO, Random Forest, Boruta, Decision Tree, XGBoost, and GBM) was utilized to pinpoint the core therapeutic target. To verify direct molecular engagement, molecular docking, molecular dynamics (MD) simulations, and Surface Plasmon Resonance (SPR) assays were performed. Finally, the functional mechanism of the identified target in CDDP resistance was validated in vitro using parental SKOV3 and resistant SKOV3/CDDP cell lines. Results scRNA-seq analysis revealed TP target genes are preferentially enriched in highly genomically unstable malignant epithelial cells. This subpopulation showed an aggressive intercellular communication profile, profound dependence on extracellular matrix (ECM) signals, and dominant secretion of the chemoresistance-related cytokine osteopontin (SPP1). Furthermore, the ML pipeline consistently pinpointed the proto-oncogene JUN as the core therapeutic target. Experiments confirmed TP efficiently suppressed aberrant c-Jun overexpression. Targeted JUN knockdown restored CDDP sensitivity, while its overexpression antagonized the synergistic cytotoxic and apoptotic effects of TP and CDDP. Ultimately, TP reverses CDDP resistance in PROC by downregulating JUN, dismantling intracellular pro-survival networks, and disrupting pro-tumorigenic crosstalk. Conclusion In conclusion, TP reverses CDDP resistance in PROC by downregulating JUN, which dismantles intracellular pro-survival networks. Integrating scRNA-seq and ML provides an accurate paradigm for deciphering botanical pharmacology, laying a strong foundation for the future development of TP-based therapies tailored for PROC.