Impact of a phosphorothioate oligodeoxynucleotide MCP-1 on NF-kappaB, AP-1, SP1 and NF-kappaB, and AP-1 subunit composition in human pulmonary endothelial cells.

Abstract

Phosphorothioate oligodeoxynucleotides (PS-ODN) are widely used prototypic antisense oligomers for sequence-specific suppression of normal and diseased gene expression. As polyanionic molecules, however, PS-ODN may also evoke nonsequence-specific side effects. The objective of the present study was to evaluate the impact of PS-ODN treatment of human pulmonary artery endothelial cells (HPAEC) and microvascular endothelial cells of the lung (HMVEC-L) on the cellular pool of the transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) as well as Sp1, using gel shift assays. In addition, by performing supershift assays, we investigated whether antisense treatment of endothelial cells affected the subunit composition of NF-kappaB and AP-1. Our data show that pretreatment of HPAEC and HMVEC-L with PS-ODN doses ranging from 50 to 5000 nM did not affect the total NF-kappaB, AP-1, or Sp1 pool in tumor necrosis factor-alpha (TNF-alpha)-activated endothelial cells (EC) or the subunit composition of the transcription factors NF-kappaB and AP-1. These findings suggest that putative nonsequence-specific effects of PS-ODN are not due to interactions of these oligomers with the transcription factors NF-kappaB, AP-1, or Sp1, at least in the EC type, a common target in transfection studies.