We employed the human glioblastoma cell line LN229 as a model system. Cells were treated with rapamycin (200 nM, 12 h) to induce autophagic stress, and acetylproteomic profiling was performed using high resolution LC MS/MS combined with immunoaffinity enrichment of acetylated peptides. We quantified 450 acetylated peptides corresponding to 333 proteins after normalization analysis when we applied a cut-off peptide abundance of 1.2-fold (log2). Interestingly, the acetylation of the majority of proteins, 439 peptides from 324 proteins, decreased after rapamycin treatment. Our dataset provides a comprehensive resource for understanding how acetylation integrates metabolic and epigenetic signals to sustain glioblastoma cell fitness under autophagic pressure. The findings may offer novel targets for combinatorial therapies that disrupt this adaptive acetylation network.