Impact of a Phosphorothioate Oligodeoxynucleotide MCP-1 on NF-κB,AP-1,SP1 and NF-κB,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, PSODN 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-κB (NF-κB) 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-κB 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-κB, AP-1, or Sp1 pool in tumor necrosis factor-α (TNF-α)-activated endothelial cells (EC) or the subunit composition of the transcription factors NF-κB 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-κB, AP-1, or Sp1, at least in the EC type, a common target in transfection studies.