Inhibited Expression of Wnt5a in H157 Cell by RNA Interference

Objective To construct PAVU6-Wnt5a/siRNA expression plasmid and inhibit its function in H157 cells.Methods Human siwnt5a was subcloned into PAVU6+27, DH10B E.col.i was transformed with the recombinant plasmid, and the ampicillin-resistant clones were identified by double digestion with HindⅢ and BamHⅠ and DNA sequencing. H157 cells were transfected with the identified PAVU6-siWnt5a and PAVU6+27 by Lipofectamine 2000.Results The DNA fragment obtained by double digestion and DNA sequencing showed that siWnt5a was inserted into the PAVU6+27 in correct orientation and reading frame. G418-resistant colonies of H157 cells were obtained. The expression of Wnt5a protein decreased by Western blotting.Conclusion PAVU6-siWnt5a expression plasmid was constructed and inhibited obviously the expression of Wnt5a in H157 cells, which help to further study the carcinogenesis and metastasis role in non-small cell lung cancer of this protein.