The role of C60 nanoparticle on apoptosis and autophagy in Hmuna Malignant Glioma

Glioblastoma is one the most aggressive types of astrocytoma, resistant chemo- and radio-therapy. Nanoparticles C60fullerene are the power antioxidant and demonstrate anticancer activity. In contrast to many chemotherapy agents, this fullerene absolutely non-toxic in wide range of concentrations. Pristine C60 fullerene is a promising candidate for many biomedical applications. Despite the fact that, the effect of C60 fullerene in cancer has been extensively studied, the potential regulation of between autophagy and apoptosis has not been addressed in glioblastoma. Molecular mechanism of crosstalk between autophagy and apoptosis in glioma cell line remain unknown. Therefore, we investigated the effect of C60 fullerene on p53/caspase-9/caspase-3/PARP and autophagy pathway on human glioblastoma astrocytoma cell. Glioblastoma U-373 cells were treated with different concentrations (0.5, 1 and 2 µM) C60 fullerene for 24 hours. Cell proliferation and viability were detected by MTT assay. ROS levels were measured by ROS/RNS and DCFD assay respectively. Changes of p53, Caspase-9, Caspase-3, PARP, p21, Beclin1 and LC3protein expressions were analysed by western blotting .Results of MTT assay showed that C60 fullerene significantly decreased cell proliferation and cell viability. All doses C60 fullerene treatment increased ROS and NO levels in U373 cell line. The expression of Beclin-1 and ratio of LC3-II/LC3-I was significantly induced by treatment of C60 fullerene. We showed that apoptosis was significantly upregulated in U373 stimulated with C60 fullerene induced apoptosis and increased the activity of caspase-9/3 and p53. We found that C60 fullerene induced cell cycle arrest, p53-dependent upregulation of p21 and activated PARP in human malignant glioma U-373 cell line after 24-hour treatment. C60 fullerene inhibited the proliferation and induced apoptosis and autophagy in U373 cells in a dose-dependent manner. Overall, our findings have suggested that water-soluble C60 fullerene induces cell cycle arrest, apoptosis, and autophagy in U373 cells through inducing p21/PARP, Beclin1/LC3 and activating p53/caspase-9/caspase-3 pathway signaling pathways. Our findings shed light on the effect of C60 fullerene in glioma cancer cells. Key words: C60 fullerene, glioma U-373, apoptosis and autophagy.