Abstract: Objective:To investigate differentially expressed protein profiles in B 16-F 10grafted melanoma and its me-tastasis in the lung in order to identify molecular markers of melanoma metastasis. Methods:Differentially expressed pro-teins in B16-F 10grafted melanoma and its metastatic lesion in the lung were isolated and identified by fluorescence two-di -mensional differential gel electrophoresis (2D-DIGE) coupled with matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS). Some of identified proteins were further confirmed by Real-time PCR analysis. Results: High resolutional images of differential gel electrophoresis were obtained and9 of 30differentially expressed proteins (|Ratio|≥2, P<0.01) were identified by MALDI-TOF-MS. The expression of Myoglobin (MB), vimentin (VIM), phosphoglycer -ate kinase 1 (PGK 1), Triosephosphate isomerase (TPI or TIM), heavy-chain binding protein(BiP), α-enolase, β-actin, γ-ac-tin, and laminin-binding protein were up-regulated in the experimental group compared with the control group. These proteins were involved in the cytoskeletal formation, glycolysis and so on. Real-time PCR analysis showed up-regulation of mRNA expression of PGK1 and TPI in the experimental group (P=0.001 and 0.003 ), which was in consistent with the re -sults of proteomic analysis.Conclusion:A variety of abnormally expressed proteins contribute to the metastasis of mice mel -anoma. Glycolytic enzymes PGK 1 and TPI may be involved in this process.