Abstract: Objective To investigate the inhibitory effect of small interfering RNA (siRNA) on expression of survivin in human melanoma M14 cells.Methods DNA templates for siRNA against survivin gene were chemically synthesized,and cloned into pRNAT-H1.1/neo plasmid.Human melanoma M14 cells were transfected with the recombinant plasmid pRNAT-H1.1/neo-survivin,and with empty plasmid as a negative control.The expression of survivin mRNA and protein were assessed by RT-PCR and Western blot analysis,respectively.Results The pRNAT-H1.1/neo-survivin siRNA expression plasmid was constructed,and verified by restriction enzyme digestion and DNA sequencing.The expression of survivin mRNA and protein in M14 cells was significantly inhibited by survivin siRNA,and the inhibition rates were 62.3±1.5% and 53.6±0.9%,respectively.Conclusions These results indicate that a siRNA expression plasmid against survivin gene,pRNAT-Hl.1/neo-survivin,is successfully constructed,and can inhibit the expression of survivin in human melanoma M14 cells.

Key words: Survivin, RNA, small interfering, Plasmids Melanoma