Isofuranodiene synergizes with Temozolomide in inducing glioma cells death

Background: Glioblastoma multiforme (GBM) is the most common and deadly brain form of tumor. GBM exhibits high resistance to the standard treatment consisting of temozolomide (TMZ) combined with radiotherapy. Isofuranodiene (IFD) is a bioactive sesquiterpene occurring in the essential oils obtained from Alexanders (Smyrnium olusatrum L., Apiaceae). This compound has shown a broad spectrum of antitumoral activities in different human cancer cell lines both in vitro and in vivo. However, the mechanism of action of IFD on GBM and its potential effects in combination with chemotherapeutic drugs, have not been fully elucidated. Purpose: The aim of the present study was to evaluate the anticancer effects of IFD itself and in combination with TMZ in GBM. Methods: Sulforhodamine B-based proliferation assay, cell cycle analysis and Annexin V/PI staining were carried out to determine the IFD effects on three human GBM cell lines, U87, T98, U251 and in normal human astrocyte. Modulation of protein expression levels was determined by western blot analysis. Reactive oxygen species (ROS) production was evaluated by cytofluorimetry. Moreover, the effects on cell viability of the IFD and TMZ co- administration was evaluated through the calculation of combination index (CI). Results: IFD exerted cytotoxic effects against the GBM cell lines, but not in normal cells (normal human astro- cytes). This compound induced a cell cycle blockage and a necrotic cell death depending on the increase of intracellular ROS levels. Furthermore, the synergism between IFD and TMZ was demonstrated in GBM cell lines. Conclusion: This study demonstrated the glioma selectivity of IFD and its cytotoxic properties suggesting a new strategy for the treatment of GBM in order to overcome the TMZ resistance and to reduce its side effects.