Figure 2

siRNA suppressed the heparanase expression in gastric cancer cells. Confluent gastric cancer SGC-7901 cells were seeded into 6-well plates, and transfected with different heparanase-specific siRNAs (siH1, siH2 and siH3) or scrambled siRNA (mock). The untransfected cells served as a control (No). The mRNA and protein expression of heparanase were examined by RT-PCR, real-time quantitative PCR and Western blot. (A) Forty-eight hrs post-transfection, siH3 (100 nmol/L) exerted the most efficiency in suppressing the heparanase expression, and siH1 (100 nmol/L) inhibited the heparanase expression to a lesser extent. However, transfection of siH2 (100 nmol/L) slightly influence the expression of heparanase in SGC-7901 cells. (B) The siH3 (100 nmol/L)-induced suppression of heparanase expression of SGC-7901 cells started at 24 hrs, and lasted for 120 hrs. Of note, the heparanase mRNA levels, but not the heparanase protein levels, were partially restored at 120 hrs post-transfection. (C) Transfection of different concentrations (10 nmol/L, 50 nmol/L and 100 nmol/L) of siH3 into SGC-7901 cells for 48 hrs, resulted in decrease of heparanase expression 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.