Abstract: Objective To investigate the role of BRAF^V600E in the development of melanoma,as well as in vivo and in vitro suppression of BRAF^V600E by short hairpin RNA(shRNA).Methods Previously constructed shRNAs targeted against the BRAF gene were used.Three plasmids,including two BRAF-specific shRNA plasmids,braf 1 and braf 2,and a negative control plasmid,were constructed and transfected into human melanoma cell line A375.A xenograft nude mouse model was used to examine the inhibitory effect ofshRNA in vivo.The expressions ofBRAF mRNA and protein were detected by RT-PCR and Western blotting,respectively.Results The specific shRNA inhibited the in vivo and in vitro expression of BRAF mRNA and protein;protein level was decreased by 62% in vivo and 90%/n vitro.In the mouse model,the tumorigenicity was decreased by the specific shRNA and the growth of tumor was inhibited by 62%.Conclusion The specific shRNAs suppress the mRNA and protein expression of BRAF^V600E,and this gene may play an important role in the development of melanoma.

Key words: shRNA, Melanoma, experimental, Genes, BRAF, RNA interference