Objective:To construct small hairpin RNA (shRNA)expression plasmids which can effectively knock down Ezrin protein expression

and to explore carrier-mediated RNA interference technology for the preventive treatment of osteosarcoma lung metastases. Methods:According to the criteria of designing small hairpin RNA(siRNA)

two villin2(coding gene of Ezrin protein)shRNA template DNA sequences were designed and synthesized. The annealed shRNA template was inserted into expression vector pGenesil-1. The positive plasmids were identified by enzyme digestion and DNA sequencing. Then the plasmids were transfected into rat osteosarcoma UMR106 cells with liposome mediation

and the expressions of Ezrin mRNA and its protein were detected by RT-PCR and Western blot. Results:Two shRNA expression plasmids for Ezrin were constructed successfully. After the transfection

the Ezrin expressions in rat osteosarcoma UMR106 cells were significantly decreased in both mRNA and protein levels. Conclusion:The shRNA expression plasmid for Ezrin can significantly inhibit Ezrin protein expression in rat osteosarcoma UMR106 cells

this studyset up a foundation for experiments of prophylactic treatment of osteosarcoma pulmonary metastasis.