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Objective: To investigate the possible correlation between osteoglycin expression and gelatinases activity of tumor cells.Methods: Eukaryotic expression plasmid and antisense expression plasmid of osteoglycin were constructed, and transfected into mouse hepatocarcinoma Hca-F cells and Hca-P cells, respectively. RT-PCR and West blot were employed to analyze osteoglycin expression in tumor cells. Zymographic analysis was used to observe the possible correlation between osteoglycin expression and gelatinases activity of tumor cells cultured under different conditions. Results: High expression of osteoglycin after transfection of osteoglycin gene attenuated secretion of gelatinases in Hca-F cells cultured with extract of lymph node (P<0.05). However, transfection of osteoglycin gene into Hca-F cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Effective suppression of osteoglycin expression in Hca-P cells by osteoglycin shRNA resulted in enhanced activity of gelatinases in Hca-P cells cultured with extract of lymph node (P<0.05). However, inhibition of osteoglycin expression in Hca-P cells failed to influence gelatinases activity of tumor cells cultured with extracts of liver and spleen or in DMEM medium. Conclusion: Osteoglycin expression influences gelatinases activity of murine hepatocarcinoma cells cultured with extract of lymph node. Regulation of gelatinases activity might be one of the mechanisms by which osteoglycin contribute to lymphatic metastasis suppression.