Construction of eukaryotic expression vector of siRNA specific for osteopontin and characterization of its efficiency in GC9811 gastric carcinoma cell line

AIM: To construct the small interfering RNA (siRNA) eukaryotic expression vector specific for human OPN gene and to observe its silencing effect on OPN gene. METHODS: The expression vectors of mU6pro/OPN-siRNA1 and mU6pro/OPN-siRNA2 were constructed by gene recombination and then were transfected by liposome-mediated transfection into the GC9811 gastric carcinoma cell line with high metastasis potential recently established in our laboratory. At 72 h after transfection, the expression of OPN in the levels of mRNA and protein was detected by RT-PCR and Western-blotting. RESULTS: The eukaryotic expression vectors of mU6pro/OPN-siRNA1 and mU6pro/OPN-siRNA2, which down-regulated mRNA and protein of OPN at 72 h after transfection, were successfully constructed. mU6pro/OPN-siRNA1 was more effective than the other one. CONCLUSION: Eukaryotic expression vector of siRNA specific for OPN is successfully constructed, which lays the basis for its application in the treatment of gastric carcinoma.