Collectively, we identified for the first time that TGF-β1 stimulated NOX4-dependent ROS expression through the Smad signal pathway, which induced metabolic reprogramming and EMT in glioblastoma and showed an uncharacterized link between tumor microenvironment, metabolism, and EMT in glioblastoma holds promising strategies for cancer therapy. The gene discussed is TGFB1; the disease is glioblastoma.