Malignant melanoma still remains to be a serious health threat.  Overexpression of focal adhesion kinase (FAK) in melanoma has suggested that FAK could be a promising target for therapeutic intervention.  To further investigate the function of FAK in melanoma, FAK expression was down-regulated by stable transfection of plasmid harboring FAK small interfering RNA (siRNA) into melanoma cell line.  Two stable cell lines, F10-siFAK and F10-control, have been constructed and screened.  Compared with the F10-control, both the mRNA and protein levels of FAK decreased significantly, and the cell cycle of F10-siFAK was arrested at G₁ phase.  Furthermore, the tumor growth rate of F10-siFAK cells was notably slower than that of F10-control in in vivo tumor models.  These results show that FAK is an important regulatory gene in melanoma.  The stable FAK-knockdown melanoma cell line is an useful tool for further investigation of FAK’s function in the progression of melanoma, and also an effective means of drug screening for anti-melanoma therapeutics.