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. 2018 Mar 1;197(5):665-668.
doi: 10.1164/rccm.201706-1304LE.

Downregulation of MicroRNA-126 Augments DNA Damage Response in Cigarette Smokers and Patients with Chronic Obstructive Pulmonary Disease

Koralia E Paschalaki  1 , Anna Zampetaki  2 , Jonathan R Baker  1 , Mark A Birrell  1 , Richard D Starke  1 , Maria G Belvisi  1 , Louise E Donnelly  1 , Manuel Mayr  2 , Anna M Randi  1 , Peter J Barnes  1
Affiliations

Downregulation of MicroRNA-126 Augments DNA Damage Response in Cigarette Smokers and Patients with Chronic Obstructive Pulmonary Disease

Koralia E Paschalaki et al. Am J Respir Crit Care Med. .
No abstract available

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Figures

Figure 1.
Figure 1.
MicroRNA-126 (miR-126) is downregulated and enhances ATM (ataxia-telangiectasia mutated) activation in smokers and patients with chronic obstructive pulmonary disease (COPD) and in an in vivo model of subchronic cigarette smoke exposure. (A) Activation of ATM was assessed by measuring phosphorylation of the ATM/ATR substrate by immunofluorescence staining (n = 3). Deep red anthraquinone 5 (DRAQ5) was used for nuclear staining. The number of nuclear foci per nucleus was quantified with Volocity software in at least five optic fields (z-stack images). An increased number of cells with focal nuclear staining of phospohrylated ATM/ATR (p-ATM/ATR) was observed in patients with COPD compared with nonsmokers. Arrows show nuclei with increased number of nuclear foci (scale bars, 20 μm). (B) We measured miR-126 by TaqMan real-time PCR in blood outgrowth endothelial cells from nonsmokers (n = 7), healthy smokers (n = 6), and patients with COPD (n = 5). RNU44 was used for normalization. (C) Human umbilical vein endothelial cells were treated with specific anti–miR-126 inhibitor versus control for 72 hours (baseline conditions, no growth factors). mRNA levels for PIK3R2 (phosphoinositol-3 kinase regulatory subunit 2; also known as p85-β) and SPRED1 (Sprouty-related EVH1 domain containing 1) (positive controls for miR-126 inhibition) and for ATM were measured by real-time PCR. ATM protein levels were quantified by Western blotting. GAPDH and α-tubulin were measured for normalization. (D–F) Male C57BL/6 mice were challenged for 28 days with cigarette smoke (for 1 hour twice daily, 4 hours apart) or ambient air. (D) miR-126 was measured by TaqMan real-time PCR in lung tissue from mice exposed to cigarette smoke (n = 6) versus control mice lung tissue (n = 5). Small nucleolar RNA135 (snoRNA135) was measured for normalization. (E and F) ATM mRNA levels were measured by real-time PCR and activation of ATM (phospho-ATM), and ATM protein levels were measured by Western blotting in lung tissue from mice exposed to cigarette smoke (n = 6) versus control mice (n = 6). GAPDH and α-tubulin were measured for normalization. (G) miR-126 was measured by TaqMan real-time PCR in lung epithelial cells from nonsmokers (n = 6) and patients with COPD (n = 6). RNU44 was used for normalization. Data presented are means ± SEM. *P < 0.05; **P < 0.01. ATR = ataxia telangiectasia and Rad3-related protein.
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References

    1. Aoshiba K, Zhou F, Tsuji T, Nagai A. DNA damage as a molecular link in the pathogenesis of COPD in smokers. Eur Respir J. 2012;39:1368–1376. - PubMed
    1. Childs BG, Durik M, Baker DJ, van Deursen JM. Cellular senescence in aging and age-related disease: from mechanisms to therapy. Nat Med. 2015;21:1424–1435. - PMC - PubMed
    1. Paschalaki KE, Starke RD, Hu Y, Mercado N, Margariti A, Gorgoulis VG, Randi AM, Barnes PJ. Dysfunction of endothelial progenitor cells from smokers and chronic obstructive pulmonary disease patients due to increased DNA damage and senescence. Stem Cells. 2013;31:2813–2826. - PMC - PubMed
    1. Fish JE, Santoro MM, Morton SU, Yu S, Yeh RF, Wythe JD, Ivey KN, Bruneau BG, Stainier DY, Srivastava D. miR-126 regulates angiogenic signaling and vascular integrity. Dev Cell. 2008;15:272–284. - PMC - PubMed
    1. Jansen F, Yang X, Hoelscher M, Cattelan A, Schmitz T, Proebsting S, Wenzel D, Vosen S, Franklin BS, Fleischmann BK, et al. Endothelial microparticle-mediated transfer of MicroRNA-126 promotes vascular endothelial cell repair via SPRED1 and is abrogated in glucose-damaged endothelial microparticles. Circulation. 2013;128:2026–2038. - PubMed

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