硫代寡脱氧核苷酸MCP-1对人肺内皮细胞NF-κB、AP-1、SP1和NF-κB以及AP-1亚基组成的影响

U. Maus, W. Seeger, J. Lohmeyer
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引用次数: 3

摘要

磷酸硫代寡脱氧核苷酸(PS-ODN)是一种被广泛应用于序列特异性抑制正常和病变基因表达的典型反义低聚物。然而,作为多阴离子分子,PSODN也可能引起非序列特异性副作用。本研究的目的是利用凝胶转移法评估PS-ODN处理人肺动脉内皮细胞(HPAEC)和肺微血管内皮细胞(HMVEC-L)对转录因子核因子-κB (NF-κB)和激活蛋白-1 (AP-1)以及Sp1的细胞池的影响。此外,通过supershift实验,我们研究了内皮细胞的反义处理是否会影响NF-κB和AP-1的亚基组成。我们的数据表明,50 ~ 5000 nM的PS-ODN预处理HPAEC和hvec - l不影响肿瘤坏死因子-α (TNF-α)激活的内皮细胞(EC)中NF-κB、AP-1或Sp1库的总量,也不影响转录因子的亚基组成。
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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
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 transcri...
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