Suberoylanilide hydroxamic acid (SAHA), as a pan HDAC inhibitor, has been approved for clinical use and it has been demonstrated to treat solid tumor. Nevertheless, the mechanism underlying the therapeutic effects of SAHA on tumors are still not fully understood. Protein phosphorylation is one of the most important regulatory events in cells, regulating primary biological processes, such as cell division, growth, migration, differentiation, and intercellular communication. The impact of SAHA treatment on cellular phosphorylation and signaling pathways might contribute to our understanding on its therapeutic mechanism. For this purpose, by combining tandem mass tags(TMT)-based quantitative proteomics and titanium dioxide-based phosphopeptide enrichment, we for the first time comprehensively identified and quantified protein phosphorylation in NPC cells toward SAHA treatment. we identified 7430 phosphorylation sites on 2456 phosphoproteins in the NPC cell line 5-8F, of which 1176 phosphorylation sites on 528 phosphoproteins were significantly expressed upon SAHA treatment. Biological analysis show that SAHA can influence several biological processes including mRNA/DNA processing, cell cycle. Furthermore, signaling pathway analysis demonstrate that SAHA can activate cell cycle regulation related pathway such as p53 signalling pathway, and inactivate energetic pathway such as AMPK pathway etc. Overall, our study presents evidence at a phosphoproteomic level that SAHA can inhibit NPC cell growth by regulating the cell cycle and cell energy metabolism. Our findings will contribute to elucidation of the mechanisms through which SAHA exerts its anticancer effects.