NF-κB regulation of endothelial cell function during LPS-induced toxemia and cancer
Tatiana Kisseleva, … , Jan Kitajewski, Christian Schindler
Tatiana Kisseleva, … , Jan Kitajewski, Christian Schindler
Published November 1, 2006
Citation Information: J Clin Invest. 2006;116(11):2955-2963. https://doi.org/10.1172/JCI27392.
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Research Article Vascular biology

NF-κB regulation of endothelial cell function during LPS-induced toxemia and cancer

Abstract

The transcription factor NF-κB is an important regulator of homeostatic growth and inflammation. Although gene-targeting studies have revealed important roles for NF-κB, they have been complicated by component redundancy and lethal phenotypes. To examine the role of NF-κB in endothelial tissues, Tie2 promoter/enhancer–IκBαS32A/S36A transgenic mice were generated. These mice grew normally but exhibited enhanced sensitivity to LPS-induced toxemia, notable for an increase in vascular permeability and apoptosis. Moreover, B16-BL6 tumors grew significantly more aggressively in transgenic mice, underscoring a new role for NF-κB in the homeostatic response to cancer. Tumor vasculature in transgenic mice was extensive and disorganized. This correlated with a marked loss in tight junction formation and suggests that NF-κB plays an important role in the maintenance of vascular integrity and response to stress.

Authors

Tatiana Kisseleva, Li Song, Marina Vorontchikhina, Nikki Feirt, Jan Kitajewski, Christian Schindler

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Figure 1

Generation of transgenic mice.

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Generation of transgenic mice. (A) The transgenic construct consisted of...
(A) The transgenic construct consisted of the Tie2 promoter, dominant negative FLAG-tagged sIkBαS32A/S36A, hGH polyadenylation cassette (hGH-pA), and the Tie2 enhancer. Restriction sites for SalI (SI), HindIII (H3), ClaI (CI), XbaI (XI), and NcoI (NI) are shown. (B) RT-PCR analysis for sIκBα expression in RNA from transgenic (line 2) and WT heart (H), lung (Lu), liver (Li), aorta (Ao), spleen (Sp), thymus (Th), and BM. The PCR product was visualized either by UV fluorescence (upper 2 panels) or hybridization with IκBα or GAPDH [32P]dCTP-radiolabeled probes (lower 2 panels). One set of BM was harvested from mice 7 days after LPS stimulation (2 μg/g). (C) IκBα and sIκBα protein expression was evaluated by a sensitive immunoblotting (anti-FLAG M5 mAb; Sigma-Aldrich)/immunoprecipitation (IκBα polyclonal Ab [pAb]; Santa Cruz Biotechnology Inc.) assay in lung, splenic, and thymic extracts prepared from transgenic mice (line 4). Extracts prepared from HEK 293T cells transiently transfected with the sIκBα expression vector served as a positive control. A nonspecific (NS) band (recognized by M5 mAb) and size standards are indicated.