Abstract 7459: The anticancer effects of brusatol are mediated by PPAR-γ/ME1-driven glycolysis in intrahepatic cholangiocarcinoma

Abstract Background: Intrahepatic cholangiocarcinoma (ICC) is a malignant tumor characterized by its dense fibrotic microenvironment, which is associated with chemotherapy resistance and poor outcome. Hence, developing novel strategies to improve the outcome of ICC is essential. Brusatol, a steroidal molecule isolated from Brucea javanica, exhibits various biological activities, including anti-inflammatory and anticancer effects. Although the molecular mechanisms underlying the anti-cancer effects of brusatol in ICC remain unclear, the present study aims to investigate its therapeutic potential in ICC cells. Methods: We conducted a series of in vitro experiments on human ICC cell lines (HCCC-9810 and HuCCT1) to assess the impact of brusatol on cell proliferation and glycolysis regulation. Transcriptomic sequencing and pathway enrichment analysis was performed to identify specific signaling pathways associated with brusatol treatment in ICC cells. Furthermore, the pathway was further validated through a series of mechanistic studies. Results: Brusatol significantly inhibited the proliferation, migration, and invasion of ICC cells in a concentration-dependent manner (p < 0.05). Glycolytic activity was markedly suppressed by brusatol, as evidenced by a reduction in glucose consumption, ATP production, and lactate secretion in ICC cells. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay confirmed the downregulation of glycolysis-related genes, including GLUT1, HK2, PFK1, and LDHA. RNA sequencing and pathway enrichment analysis suggested that brusatol inhibits the PPAR-γ/ME1 signaling pathway (p < 0.05), which likely regulates glycolysis in ICC cells. Subsequently, RT-qPCR and western blotting results corroborated the RNA-seq findings. Interfering with PPAR-γ expression via siRNA or inhibiting the PPAR-γ pathway with Rosiglitazone resulted in a marked reduction in glycolytic activity (p < 0.01), further reinforcing the critical role of the PPAR-γ/ME1 signaling axis in the regulation of glycolysis. These findings underscore the potential of brusatol as a therapeutic agent that effectively disrupts glycolytic metabolism in ICC by targeting the PPAR-γ/ME1 pathway. Conclusion: Our findings provide compelling evidence that the PPAR-γ/ME1 signaling pathway mediates the anti-cancer effects of brusatol, supporting its potential as a safe and effective integrative treatment approach for ICC. This potential offers hope for the development of new and improved treatments for ICC. Citation Format: Xiaoqing Wang, Caiming Xu, Kai Luo, Yuhan Fang, Goel Ajay, Guixin Zhang. The anticancer effects of brusatol are mediated by PPAR-γ/ME1-driven glycolysis in intrahepatic cholangiocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 7459.