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Nitric oxide upregulation of caspase-8 mRNA expression in lung endothelial cells: role of JAK2/STAT-1 signaling

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Abstract

We recently reported that nitric oxide (NO) modulates expression of multiple genes associated with apoptotic pathways, including expression of caspase-8. The objective of the present study is to determine whether the NO-induced expression of the caspase-8 gene is regulated via signal transducers and activators of transcription-1 (STAT-1) signaling. The confluent monolayers of pulmonary artery endothelial cells (PAEC) were incubated with or without (control) 1 mM NOC-18, a NO donor, at 37°C for 0–24 h. In some experiments PAEC were pretreated with a Janus kinase (JAK-2) inhibitor, AG490 (20 μM). Exposure of PAEC to NO-increased relative levels of caspase-8 mRNA as determined using quantitative real time PCR. Relative levels of phosphorylated STAT-1 at Serine (Ser)-727, but not total STAT-1 expression in NO-exposed cells, were upregulated significantly compared to control cells. AG490 attenuated NO-induced phosphorylation of STAT-1 at Ser 727 and expression of caspase-8 mRNA, suggesting JAK2 plays a role in the induction of caspase-8 mRNA. The promoter of caspase-8 has four γ-activated sequence (GAS) and two interferon-stimulated response element (ISRE) transcription factor-binding sites. NO enhanced the STAT-1 binding activity to GAS/ISRE. Suppression of STAT-1 expression attenuated NO-induced elevation of caspase-8 mRNA. These studies demonstrate that a NO-dependent increase in caspase-8 mRNA levels is associated with phosphorylation of STAT-1 at Ser-727 and STAT1 binding to the caspase-8 promoter in cultured PAEC.

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Acknowledgments

The authors thank Ms. Gina. Eubanks and Mrs. Di-hua He for technician assistance and Mr. Bert Herrera for tissue culture assistance. This work was supported in part by the Medical Research Service of the Department of Veterans Affairs (JMP, ERB), by NIH grant HL-67886 (JMP), and by a Scientist Development Award from the American Heart Association of Florida (JZ).

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Authors and Affiliations

  1. Department of Medicine, University of Florida College of Medicine, Gainesville, FL, 32608-1197, USA

    Liuzhe Li, Jianliang Zhang, Bilian Jin, Edward R. Block & Jawaharlal M. Patel

  2. Research Service (151), Malcom Randall Department of Veterans Affairs Medical Center, 1601 S.W. Archer Road, Gainesville, FL, 32608-1197, USA

    Edward R. Block & Jawaharlal M. Patel

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  1. Liuzhe Li
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  2. Jianliang Zhang
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Correspondence to Jawaharlal M. Patel.

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Li, L., Zhang, J., Jin, B. et al. Nitric oxide upregulation of caspase-8 mRNA expression in lung endothelial cells: role of JAK2/STAT-1 signaling. Mol Cell Biochem 305, 71–77 (2007). https://doi.org/10.1007/s11010-007-9529-z

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  • DOI: https://doi.org/10.1007/s11010-007-9529-z

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