Abstract
Objective: To investigate the antitumor activity of the natural compound Furanodiene in glioblastoma multiforme (GBM), evaluate its synergistic effect with temozolomide (TMZ), and assess its potential to enhance antitumor immune responses. Methods: LN229 and U251 glioma cells, as well as patient-derived glioma stem-like cells (GSCs), were treated with Furanodiene alone or in combination with TMZ. Cell proliferation, apoptosis, and neurosphere formation were evaluated in vitro. For in vivo experiments, orthotopic glioma models were established in nude and C57BL/6 mice. Tumor burden was monitored via bioluminescence imaging, and survival was evaluated using the Kaplan-Meier analysis. Tumor sections were examined by immunofluorescence (Ki67, TUNEL), while immune responses were assessed via flow cytometry. Tumor rechallenge assays were performed to evaluate immune memory. Results: Furanodiene inhibited cell proliferation and induced apoptosis in a dose-dependent manner. Combined treatment with TMZ further enhanced cytotoxicity and suppressed neurosphere formation. In vivo, Furanodiene suppressed tumor growth, increased apoptosis, decreased Ki67 expression, and prolonged survival. Flow cytometry revealed increased CD8+ T cell infiltration and TNF-α expression in tumor tissues. Rechallenge experiments confirmed resistance to tumor regrowth, indicating long-term immune memory. Conclusions: Furanodiene exerts potent antitumor effects and potentiates TMZ efficacy in GBM. Moreover, it enhances T cell activation and induces durable immune protection, supporting its potential as both a therapeutic and immunomodulatory agent.