Gliomas are primary intracranial tumors that cause considerable morbidity and mortality. The effect of interleukin 4-induced gene-1 (IL4I1) on the progression of various diseases has been demonstrated to be significant. However, the specific molecular mechanisms of how IL4I1 contributes to the progression and the epithelial-mesenchymal transition (EMT) process of glioma remain inadequately elucidated. IL4I1 expression in glioma was assessed using public datasets and immunohistochemistry. In in vitro experiments, IL4I1 expression was quantified through real-time quantitative PCR and western blotting. The effects of IL4I1 knockdown on the malignant phenotypes of glioma cells were investigated through in vitro studies. The evaluation of biomarkers associated with EMT and the Janus-activated kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway was conducted using western blotting and immunofluorescence assays after IL4I1 knockdown. A xenograft tumor model was established to validate the influence of IL4I1 knockdown in glioma progression. The results revealed that high expression of IL4I1 is linked to an unfavorable prognosis in human gliomas. IL4I1 knockdown effectively impeded the malignant phenotypes of glioma cells. IL4I1 knockdown induced EMT reversal, characterized by alterations in the expression levels and localization of EMT-related biomarkers. This reversal is partially mediated through the JAK2/STAT3 signaling pathway. The results of in vivo experiments confirmed that IL4I1 knockdown effectively suppressed glioma growth. Our research demonstrates that IL4I1 knockdown reverses the EMT process via JAK2/STAT3 signaling pathway and suppresses the malignant phenotypes of glioma, thereby highlighting its potential as both a prognostic marker and therapeutic target for glioma.