Figure 4

Silencing heparanase abolished the adhesion, migration and invasion of gastric cancer cells in vitro. Confluent SGC-7901 cells were seeded into 96-well or 24-well plates, and transfected with different concentrations (10 nmol/L, 50 nmol/L, and 100 nmol/L) of siH3 or scrambled siRNA (mock, 100 nmol/L). The untransfected cells served as a control (No). (A) In the adhesion assay, 48 hrs post-transfection, 2 × 10⁴ cells were inoculated into each matrigel-coated well of 96-well plates for 2 hrs, and washed three times with PBS. The results indicated that transfection of high concentrations of siH3 (50 nmo/L and 100 nmol/L), but not low concentration of siH3 (10 nmol/L) or mock, reduced the adhesion of SGC-7901 cells to the matrigel. (B) In scratch migration assay, 48 hrs post-transfection, the cells were scraped with 1-ml pipette tips (time 0). Cell migration into the wounded empty space was followed after 24 hrs and photographed. The results indicated that transfection of siH3 impaired the cellular migration in a dose-dependent manner. (C) In transwell analysis, 48 hrs post-transfection, homogeneous single cell suspensions (1 × 10⁵ cells/well) were added to the upper chambers and allowed to invade for 24 hrs. Migrated cells were stained with 0.1% crystal violet and examined by light microscopy. The results indicated that transfection of siH3 abolished the invasion of SGC-7901 cells in a dose-dependent manner. The symbols (* and **) indicates a significant (P < 0.05) and a very significant (P < 0.01) decrease from mock, respectively. Triplicate experiments were performed with essentially identical results.