Effects of heparanase gene silencing by RNA interference on invasion of gallbladder carcinoma in vitro

Objective:To investigate the effects of heparanase(HPA) gene silencing by miRNA interference on invasion and migration of GBC-SD cells of gallbladder carcinoma. Methods:Four pairs of single-stranded recombinant plasmid miRNA oligo-DNA targeting human HPA gene sequence were synthesized and transfected into GBC-SD cells by lipofectamine^TM 2000. HPA mRNA expression was detectede by RT-PCR before and after transfection, the recombinant plasmid which showed the best silencing effect for further experiments was selected. Transwell invasion assays and wound scarification assays were performed to examine the migration and invasive ability of GBC-SD cells. Results:Recombinant plasmid pcDNA6.2-GW/EmGFP-miR-HPA-2 showed the best silencing effects. The migration distance in wound scarification assays of GBC-SD cells transfected with recombinant plasmid pcDNA 6.2-GW/EmGFP-miR-HPA-2 at 12h,24h was significant shorter than negative control and blank groups(P<0.05-P<0.01). The invasive ability of GBC-SD cells transfected with pcDNA6.2-GW/EmGFP-miR-HPA-2 was decreased significantly when compared with negative control and blank groups(P<0.01). Conclusions:The GBC-SD cells transfected with recombinant plasmid pcDNA6.2-GW/EmGFPmiR showed decreased HPA mRNA expression,as well as inhibited invasion and migration in vitro.