Curcumin, derived from the rhizome of Curcuma longa is a natural anti-cancer agent and has been shown to inhibit proliferation and survival of tumor cells. Although the anti-cancer effects of curcumin are well established, detailed understanding of the signaling pathways altered by curcumin is still unclear. In this study, we carried out SILAC-based quantitative proteomic analysis of CAL 27 cell line, a HNSCC cell line to investigate tyrosine signaling in response to curcumin. Using high resolution Orbitrap Fusion™ Tribrid™ Fourier transform mass spectrometer, we identified 627 phosphotyrosine sites mapping to 357 proteins. We observed alterations in the level of phosphorylation of 307 sites corresponding to 204 proteins upon curcumin treatment.