Laryngeal carcinoma related gene LCRG1, cloned by the laboratory using mRNA differential display, has the suppressive function to none expression LCRG1 Hep-2 cell line. Bioinformatics analysis using software showed LCRG1 may play function in cellular signal transduction. In order to further elucidate the function of LCRG1, RT-PCR and colony efficiency were used to identify whether LCRG1 expressed and had the tumor suppressive function in incubated Hep-2/LCRG1 cell lines. The results suggested LCRG1 was expressed in Hep-2/LCRG1 cell lines and had the significant suppressive proliferation ability. Hence, the total proteins of Hep-2/LCRG1 and Hep-2/pcDNA3.1(+) cell lines were separated by immobilized pH gradient(IPG)-based two-dimensional gel electrophoresis(2DGE), coupled with anti-tyrosine phosphorylated antibody immunoblotting and matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS) identifying tyrosine-phosphorylated proteins. The well-resolved, reproducible 2DGE patterns of Hep-2/LCRG1 and Hep-2/pcDNA3.1(+) cell lines were established and 13 differential tyrosine-phosphorylated proteins were identified using immunoblotting, analysis software and MALDI-TOF-MS methods. These proteins were involved in the signal transduction and cell cycle. So it was speculated that LCRG1 may be involved in the processes of cellular proliferation, metabolic pathways and apoptosis etc. and play tumor suppressive functions through regulating the phosphorylation/ dephosphorylation status of these proteins. These data will be helpful to elucidate the molecular mechanism of LCRG1 tumor suppressive function.