Aim To investigate the simultaneous effects of antisense and decoy oligonucleotides of nuclear factor (NF) κB on expression of adhesion molecules induced by tumor necrosis factor(TNF)-α in vitro. Methods Intracellular uptake of fluorescein isothiocyanate (F1TC) -labeled oligonucleotides was detected by fluorescence active cell sort (FACS) ; Adhesion molecules expression was presented by reverse transcription-polumerase chain reaction (RT-PCR) at RNA level, FACS at protein level. Results Simultaneous incubation with antisense P65 and decoy oligonucleotides (ODN) of NF-κB, the inhibitory effect of VGAM-1 was 59.0%, 41.1% and 35.2% at mRNA level, 85.5%, 44.3% and 49.2% on human vascular endothclial cells (hUVEC) surface compared with control group, decoy group and antisense group. The inhibitory effect of simultaneous incubation with antisense and decoy oligonucleotides was stronger than the effect of incubation with one of them. Conclusion It was demonstrated that a simultaneous administration of antiscnse and decoy oligonucleotides of NF-κB was effective and necessary if a sufficient inhibition of NF-κB mediated activities should be achieved.