In order to study the relationship between sphingomyelin synthase(SMS) activity and sphingomyelin(SM) metabolism, we transfected HEK293 cells with SMS1 or SMS2, two isoforms and SMS, expression vector. The expression levels of SMS were monitored by SMS activity assay, and cellular and medium SM levels were determined by an established method. The results show that SMS1 overexpression significantly increased SMS activity(65%), cellular and medium SM levels(58% and 46%, p<0.01, respectively) in comparison with control cells. Whereas, SMS2 overexpression also significantly increased SMS activity(13%) and SM levels in the cells(33%, p<0.05) or in the medium(29%, p<0.05) in comparison with controls. The results show that SM levels can be regulated by SMS1 or SMS2, suggesting that the monitoring SMS activity could be an alternative approach to treat atherosclerosis.