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.
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.
Department of Internal Medicine, Justus-Liebig-University, Giessen, Germany. Ulrich.A.Maus@med.uni-giessen.de
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D001483:Base Sequence; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors* / Q000235:genetics*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000098:blood supply; D016328:NF-kappa B / Q000737:chemistry / Q000378:metabolism; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D021122:Protein Subunits; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013873:Thionucleotides / Q000235:genetics / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000737:chemistry / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018932:Chemokine CCL2; D016328:NF-kappa B; D020319:Oligodeoxyribonucleotides, Antisense; D021122:Protein Subunits; D016329:Sp1 Transcription Factor; D013873:Thionucleotides; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha