Herein, we reported for the first time that fractionated IR (fraction-dependent manner as to the clinical daily used dose: 2 Gy per day) induced GSCs with the stemness phenotype by p62-mediated autophagy through the Wnt/β-catenin/GSk3β/P62 axis signaling pathway in human glioma cells. The gene discussed is GSK3B; the disease is central nervous system cancer.