Objective To investigate the starvation-induced autophagy in rat C6 glioma cells and the changes of autophagy-related protein Beclin-1. Methods Rat C6 glioma cells of the logarithmic growth phase were cultured with amino acid-free EBSS medium instead of the DMEM medium. Cells were collected at 1 h, 3 h, 6 h, 12 h, and 18 h after EBSS culture. Western blotting analysis and immunofluorescence were used to detect the specific markers of autophagic microtubule-associated protein 1 light chain 3 (LC-3) and autophagy-related protein Beclin-1, and transmission electron microscope was used to observe the autophagic body. Results LC-3 expression was found in C6 glioma cells 1 h after starvation; the expression increased with the prolongation of starvation, and reached a peak 12 h later and then began to decline. Autophagic bodies were observed under transmission electron microscopy, and autophagosomes were found in immunofluorescence. Beclin-1 expression began to increase in C6 glioma cells 1 h after starvation, reached a peak at 3 h and then decreased gradually. Increased expression of Beclin-1 was also observed by immunofluorescence. Conclusion Starvation can induce autophagy in C6 glioma cells, which is correlated with the expression of autophagy-related protein Beclin-1. This study lays a foundation for studying the autophagy mechanism in glioma cells.