Aim To investigate the inhibitory effect of antisense monocyte chemotactic protein-1 (MCP-1) expression mediated by recombinant gene transfer on the migration of monocytes into arteries in vivo, as a first step, a recombinant retroviral vector expressing antisense MCP-1 gene was constructed and its expression was observed in vitro. Methods An antisense MCP-1 recombinant vector was constructed by inserting rabbit MCP-1 cDNA into LNCX plasmid in a reversed orientation. Recombinant viral DNA was transfected into φ-2 packaging cells by lipofectamine transfecting procedure. The transiently produced viruses were used to transfer PA317 amphotropic packaging cells. G418 resistant positive PA317 clones were isolated and expanded. Virus surpernatant from G418 resistant PA317 cells was used to determine virus titer and to infect aortic smooth muscle cells. After the selection with G418 medium, the integration and expression of recombinant gene in cells were measured. Results The highest retroviruses' titer was 5.6×10 7 CFU/L. PCR result showed that recombinant viruses were integrated into genome of the infected NIH3T3 cells and cultured smooth muscle cells. RNA slot blot analysis showed that antisense MCP-1 mRNA expressing was detected in the infected smooth muscle cells. There was obvious inhibition of MCP-1 mRNA expression obtained in the infected smooth muscle cells in comparing to that the uninfected cells. Conclusions The recombinant retroviral vector containing antisense MCP-1 could inhibit target gene expression in cultured smooth muscle cells and may be used for further experimental studies in vivo.