Berberine, a quaternary amine and isoquinoline alkaloid, has been proposed to have antimetastatic effects on many types of tumor cells; however, its exact inhibitory mechanisms with bladder cancer cells remain unclear. We used berberine and siRNA technology to interfere the expression of heparanase in bladder cancer T24 cells, detected the expression of heparanase mRNA and protein by reverse transcription PCR and Western blot respectively, and investigated their effects on the migration and invasion of T24 bladder cancer cells using transwell chamber. Our results showed that both mRNA and protein of heparanase were highly expressed in human bladder cancer T24 cells and markedly downregulated by both heparanase-specific siRNA (hpa-siRNA) and berberine. The tumor cell migration assay indicated that transfection of hpa-siRNA and treatment with berberine both attenuated the migration and invasion of T24 cells. Therefore, berberine inhibits the metastasis and invasion of bladder cancer cell, possibly via blocking the heparanase expression and thus may be used clinically to reduce the recurrence of bladder cancer.