<p>Glioma is a highly invasive primary brain tumor with a poor prognosis and currently lacks effective treatment methods. Increasing evidence indicates that mitochondrial oxidative phosphorylation (OXPHOS) is crucial for the development of glioma; however, the regulatory mechanisms controlling mitochondrial protein synthesis and energy metabolism are not yet fully understood. Analysis of the TCGA and CGGA databases reveals that MRPL27 is highly expressed in glioma tissues and is significantly associated with poor patient survival. Silencing MRPL27 significantly inhibits the proliferation, migration, and tumor growth of glioma cells, while inducing cell apoptosis. Mechanistically, the absence of MRPL27 impairs the mitochondrial oxidative phosphorylation process, reduces ATP production, disrupts redox balance, and increases oxidative stress. Notably, MRPL27 deficiency specifically reduces the protein content of mitochondrial-encoded oxidative phosphorylation components without altering their transcriptional levels, indicating its role in post-transcriptional regulation of mitochondrial protein expression. In summary, these findings suggest that MRPL27 is a key regulator of mitochondrial translation and energy metabolism in glioma, and emphasize the significance of mitochondrial ribosome regulation as a potential metabolic weakness for therapeutic intervention.</p> Graphical Abstract <p></p>