Conclusively, our findings demonstrate that EGFRvIII promotes the proneural–mesenchymal transition of GBM and reduces its sensitivity to TMZ by regulating the NF-κB/ALDH1A3 axis, which providing an experimental basis for the selection of clinical drugs for GBM patients with EGFRvIII (+). Here, NFKB1 is linked to glioblastoma.